Patent Application
20250092032
The present invention relates to the field of medicine, in particular to Deoxycytidine kinase (dCK) inhibitors and their uses for treating a cancer.
Deoxycytidine kinase (dCK) is an enzyme having a crucial role in cellular division. This enzyme allows the phosphorylation of a large number of deoxyribonucleosides and their nucleoside analogs. More particularly, dCK catalyzes the 5′-phosphorylation of physiologic pyrimidines and purines, such as 2′-deoxycytosine (dC), 2′-deoxyadenosine (dA) and 2′-deoxyguanosine (dG). dCK has been observed to be predominantly expressed in a large panel of human cancer models in the Cancer Cell Line Encyclopedia (https://portals.broadinstitute.org/ccle) and was associated with certain forms of resistance to antiviral and anticancer chemotherapeutic agents. In this context, dCK has been validated as an interesting target in oncology. So far, there remains a need to develop further dCK inhibitors for the treatment of diseases and disorders for which dCK activity is implicated, such as cell proliferative diseases, particularly cancer.
In this context, the inventors have provided new compounds having the properties to bind to dCK and inhibit its activity, demonstrating thereby the therapeutic interest of such compounds in medicine, more particularly in cancer therapies.
The present invention thus provides a compound having the following formula (I):
wherein:
In a particular embodiment, the compound of formula (I) is such that:
Preferably, said compound has the following formula (I′):
in which, R1, R2, R3, R4, R5, R6, X1, X2, X3, n1, n2, and n3 are such as defined herein.
In a particular embodiment, the compound of formula (I) or (I′) is such that at least one group chosen among R1, R2, and R3 is not a hydrogen.
In a particular embodiment, the compound of formula (I) or (I′) is such that:
In a particular embodiment, the compound of formula (I) or (I′) is such that:
In a particular embodiment, the compound of formula (I) or (I′) is such that R6 represents a hydrogen.
In a particular embodiment, the compound of formula (I) or (I′) is such that n1+n2+n3 is equal or superior to 1, preferably n1+n2+n3 is 1, 2, or 3.
In a particular embodiment, the compound of formula (I) or (I′) is such that n1 is 0.
In a further particular embodiment, the compound of formula (I) or (F) is such that n1 is 1 and X1 represents a —NHCO— group.
In a particular embodiment, the compound of formula (I) or (I′) is such that n2 is 1 and X2 represents a 5-12 membered ring selected in the group consisting of a phenyl, a pyrimidinyl, a thiophenyl, a pyridinyl, a triazolyl, and an indolyl, said 5-12 membered ring is optionally substituted by at least one radical selected in the group consisting of a methoxy, a trifluoromethoxy, a halogen, a hydroxy, a methyl, and a trifluromethyl.
In a particular embodiment, the compound of formula (I) or (I′) is such that n3 is 1 and X3 represents a radical selected in the group consisting of a (C1-C6)alkyl, a (C1-C6)alkoxy, a —C(O)—, a —SO2—, a —NH—SO2—, and a —NHCO—, said radical being substituted by a piperazinyl optionally substituted by a (C1-C6)alkyl, preferably a piperazinyl substituted by a methyl.
In a further particular embodiment, the compound of formula (I) or (I′) is such that n3 is 1 and X3 represents a (C1-C6)alkyl substituted by at least one radical A as defined herein, preferably, a piperazinyl substituted by a methyl, a —N(CH3)2, a 2,6-diazaspiro[3.3]heptanyl substituted by a methyl, and a —NHSO2CH3.
In a further particular embodiment, the compound of formula (I) or (I′) is such that n3 is 1 and X3 represents a —SO2— substituted by at least one radical A as defined herein, preferably a piperazinyl substituted by a methyl, a —NR11R12 group with Rn and R12 being independently a hydrogen or a methyl group, and a morpholinyl.
In a preferred embodiment, the compound of formula (I) or (I′) is such that:
A preferred compound of formula (I) is selected in the group consisting of:
A further object of the invention is a compound as defined herein for use as a drug.
A further object is a pharmaceutical composition comprising a compound as defined herein, and a pharmaceutically acceptable excipient. In a particular embodiment, the pharmaceutical composition further comprises an inhibitor of the De Novo nucleotide biosynthesis pathway, particularly a ribonucleotide reductase inhibitor, preferably thymidine.
Another object of the invention is a compound as defined herein or a pharmaceutical composition as defined herein for use for treating a cancer, preferably a liquid cancer, more preferably acute lymphoblastic leukemia, even more preferably T-cell acute lymphoblastic leukemia.
According to the present invention, the terms below have the following meanings:
The terms mentioned herein with prefixes such as for example C1-C6, can also be used with lower numbers of carbon atoms such as C1-C2. If, for example, the term C1-C6 is used, it means that the corresponding hydrocarbon chain may comprise from 1 to 6 carbon atoms, especially 1, 2, 3, 4, 5, or 6 carbon atoms. If, for example, the term C1-C3 is used, it means that the corresponding hydrocarbon chain may comprise from 1 to 3 carbon atoms, especially 1, 2, or 3 carbon atoms.
The term “alkyl” refers to a saturated, linear or branched aliphatic group. The term “(C1-C6)alkyl” more specifically means methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, isopentyl, or hexyl.
The term “alkoxy” or “alkyloxy” corresponds to the alkyl group as above defined bonded to the molecule by an —O— (ether) bond. (C1-C6)alkoxy includes methoxy or methyloxy, ethoxy or ethyloxy, propoxy or propyloxy, isopropoxy or isopropyloxy, butoxy or butyloxy, isobutoxy or isobutyloxy, pentoxy or pentyloxy, isopentoxy or isopentyloxy, and hexoxy or hexyloxy.
The term “3-20 membered ring” corresponds to a ring having between 3 and 20 atoms. Such a term includes the term “5-12 membered ring” having between 5 and 12 atoms. The term “ring” corresponds to a mono-, bi, or tricycle, which can be saturated or unsaturated, and optionally comprises at least one heteroatom. Particularly, the term “ring” includes a cycloalkyl, a heterocycloalkyl, an aryl, and a heteroaryl.
The term “cycloalkyl” corresponds to a saturated or unsaturated mono-, bi- or tri-cyclic alkyl group comprising between 3 and 20, preferably between 5 and 12 atoms of carbons. It also includes fused, bridged, or spiro-connected cycloalkyl groups. The term “cycloalkyl” includes for instance cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.
The term “heterocycloalkyl” corresponds to a saturated or unsaturated cycloalkyl group as above defined further comprising at least one heteroatom such as nitrogen, oxygen, or sulphur atom. It also includes fused, bridged, or spiro-connected heterocycloalkyl groups.
Representative heterocycloalkyl groups include, but are not limited to dioxolanyl, benzo[1,3]dioxolyl, azetidinyl, oxetanyl, pyrazolinyl, pyranyl, thiomorpholinyl, pyrazolidinyl, piperidyl, piperazinyl, 1,4-dioxanyl, imidazolinyl, pyrrolinyl, pyrrolidinyl, piperidinyl, azepanyl, 2,6-diazaspiro[3.3]heptanyl, imidazolidinyl, morpholinyl, 1,4-dithianyl, pyrrolidinyl, oxozolinyl, oxazolidinyl, isoxazolinyl, isoxazolidinyl, thiazolinyl, thiazolidinyl, isothiazolinyl, isothiazolidinyl, dihydropyranyl, tetrahydropyranyl, tetrahydrofuranyl, and tetrahydrothiophenyl. In a preferred embodiment, the heterocycloalkyl group is azepanyl, piperazinyl, morpholinyl, piperidinyl, and 2,6-diazaspiro[3.3]heptanyl.
The term “aryl” corresponds to a mono- or bi-cyclic aromatic hydrocarbons having from 6 to 12 carbon atoms. For instance, the term “aryl” includes phenyl, naphtalenyl, or anthracenyl. In a preferred embodiment, the aryl is a phenyl.
The term “heteroaryl” as used herein corresponds to an aromatic, mono- or poly-cyclic group comprising between 5 and 14 atoms and comprising at least one heteroatom such as nitrogen, oxygen or sulphur atom. As used herein, the term “heteroaryl” further includes the “fused arylheterocycloalkyl” and “fused heteroarylcycloalkyl”. The terms “fused arylheterocycloalkyl” and “fused heteroarylcycloalkyl” correspond to a bicyclic group in which an aryl as above defined or a heteroaryl is respectively bounded to the heterocycloalkyl or the cycloalkyl as above defined by at least two carbons. In other terms, the aryl or the heteroaryl shares a carbon bond with the heterocycloalkyl or the cycloalkyl. Examples of such mono- and poly-cyclic heteroaryl group, fused arylheterocycloalkyl and fused arylcycloalkyl may be: pyridinyl, thiophenyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, tetrazolyl, oxadiazolyl, furazanyl, thiadiazolyl, tetrazolyl, benzofuranyl, thianaphthalenyl, indolyl, indolinyl, indanyl, quinolinyl, isoquinolinyl, benzimidazolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, triazinyl, thianthrenyl, benzofuranyl, dihydrobenzofuranyl, isobenzofuranyl, benzothiophenyl, isobenzothiophenyl, chromenyl, xanthenyl, phenoxanthinyl, pyrazinyl, pyridazinyl, indolizinyl, isoindolyl, indazolyl, purinyl, quinolizinyl, phtalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, pteridinyl, carbazolyl, β-carbolinyl, phenanthridinyl, acridinyl, pyrimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, furazanyl, phenoxazinyl, isochromanyl, chromanyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, indolinyl, isoindolinyl, oxazolidinyl, benzotriazolyl, benzoisoxazolyl, oxindolyl, benzoxazolyl, benzoxazolinyl, benzoxazinyl, benzothienyl, benzothiazolyl, benzodiazepinyl, benzazepinyl, benzoxazepinyl, isatinyl, dihydrobenzodioxepinyl, dihydropyridyl, s-triazinyl, oxazolyl, or thiofuranyl. In a preferred embodiment, the heteroaryl group is pyrimidinyl, thiophenyl, pyridinyl, triazolyl, and indolyl.
The term “halogen” corresponds to a fluorine, chlorine, bromine, or iodine atom, preferably a fluorine, chlorine or bromine atom, preferably a fluorine or a chlorine.
The expressions “a radical substituted by a” and “a radical substituted by at least” means that the radical is substituted by one or several groups of the list. For instance, the expression “a (C1-C6)alkyl substituted by at least one halogen, preferably a fluorine, may include a fluoromethyl (—CH2F), a difluoromethyl (—CHF2), or a trifluoromethyl (—CF3).
The expression “optionally substituted” means that the radical is not substituted or substituted by one or several groups of the list.
The “tautomers” are isomeric compounds that differ only in the position of the protons and the electrons.
The “solvates” are compounds further comprising at least one molecule of solvent. The “hydrates” are compounds further comprising at least one molecule of water. For instance, if the compound comprises one molecule of water, it corresponds to a monohydrate form. If the compound comprises two molecules of water, it corresponds to a dihydrate form.
The “pharmaceutically salts” include inorganic as well as organic acids salts. Representative examples of suitable inorganic acids include hydrochloric, hydrobromic, hydroiodic, phosphoric, di- or tri-hydrochloric, di- or tri-hydrobromic, di- or tri-hydroiodic, di- or tri-phosphoric, and the like. Representative examples of suitable organic acids include formic, acetic, trichloroacetic, trifluoroacetic, propionic, benzoic, cinnamic, citric, fumaric, maleic, methanesulfonic and the like. Further examples of pharmaceutically inorganic or organic acid addition salts include the pharmaceutically salts listed in J. Pharm. Sci. 1977, 66, 2, and in Handbook of Pharmaceutical Salts: Properties, Selection, and Use edited by P. Heinrich Stahl and Camille G. Wermuth 2002. In a preferred embodiment, the salt is selected from the group consisting of maleate, chlorhydrate, bromhydrate, and methanesulfonate. The “pharmaceutically salts” also include inorganic as well as organic base salts. Representative examples of suitable inorganic bases include sodium or potassium salt, an alkaline earth metal salt, such as a calcium or magnesium salt, or an ammonium salt. Representative examples of suitable salts with an organic base includes for instance a salt with methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris-(2-hydroxyethyl)amine. —COR may refer to —C(O)—R, —CO— may refer to —C(O)—, —CONHR may refer to —C(O)—NH—R, —NRR′ may refer to —N(R)R′, —NHCO— may refer to —NH—CO—, —O—CH2— may refer to —CH2—O—, and —CO2R may refer to —C(O)—O—R.
As used herein, the terms “treatment”, “treat” or “treating” refer to any act intended to ameliorate the health status of patients such as therapy, prevention, prophylaxis and retardation of a disease, in particular a cancer. In certain embodiments, such terms refer to the amelioration or eradication of the disease, or symptoms associated with it. In other embodiments, this term refers to minimizing the spread or worsening of the disease, resulting from the administration of one or more therapeutic agents to a subject with such a disease.
As used herein, the terms “subject”, “individual” or “patient” are interchangeable and refer to a mammal, even more preferably to a human, including adult, child, newborn and human at the prenatal stage. However, the term “subject” can also refer to non-human animals, in particular mammals such as dogs, cats, horses, cows, pigs, sheep and non-human primates, among others.
The terms “quantity,” “amount,” and “dose” are used interchangeably herein and may refer to an absolute quantification of a molecule.
As used herein, the terms “active principle”, “active ingredient”, “active pharmaceutical ingredient”, and “drug” are equivalent and refer to a component of a pharmaceutical composition having a therapeutic effect.
As used herein, the term “therapeutic effect” refers to an effect induced by an active ingredient, or a pharmaceutical composition according to the invention, capable to prevent or to delay the appearance or development of a disease or disorder, or to cure or to attenuate the effects of a disease or disorder.
As used herein, the term “effective amount” refers to a quantity of an active ingredient or of a pharmaceutical composition which prevents, removes or reduces the deleterious effects of the disease, particularly a cancer. It is obvious that the quantity to be administered can be adapted by the man skilled in the art according to the subject to be treated, to the nature of the disease, etc. In particular, doses and regimen of administration may be function of the nature, of the stage and of the severity of the disease to be treated, as well as of the weight, the age and the global health of the subject to be treated, as well as of the judgment of the doctor.
As used herein, the term “pharmaceutically acceptable excipient” refers to any ingredient except active ingredients which are present in a pharmaceutical composition. Its addition may be aimed to confer a particular consistency or other physical or gustative properties to the final product.
A pharmaceutically acceptable excipient must be devoid of any interaction, in particular chemical, with the active ingredients.
The present invention provides new compounds of therapeutic interest.
According to the invention, a compound has the following formula (I):
wherein:
According to a particular embodiment of the invention, a compound has the following formula (I):
wherein:
According to the invention, Y and Z represent independently NH, N, O, or S. Particularly, Y and Z represent independently N, O, or S. In a preferred embodiment, Y represents N and Z represents O or S. In a more preferred embodiment, Y represents N and Z represents S.
According to this more preferred embodiment, the compound has the following formula (I′):
in which, R1, R2, R3, R4, R5, R6, X1, X2, X3, n1, n2, and n3 are such as defined herein.
According to the invention, R1, R2, and R3 represent independently a radical selected in the group consisting of a hydrogen, a —NR7R8 group with R7 and R8 being independently a hydrogen or a (C1-C6)alkyl group, and a halogen. In a particular embodiment, R1 represents a hydrogen or a —NR7R8 group with R7 and R8 being a hydrogen, i.e. an amino group —NH2; R2 represents a hydrogen or a halogen such as a fluorine, preferably a hydrogen; and R3 represents a hydrogen or a —NR7R8 group with R7 and R8 being a hydrogen, preferably a —NR7R8 group with R7 and R8 being a hydrogen.
In a particular embodiment, at least one group chosen among R1, R2, and R3 is not a hydrogen.
In a more particular embodiment, R1, R2, and R3 represent independently a radical selected in the group consisting of a hydrogen, a —NR7R8 group with R7 and R8 being independently a hydrogen or a (C1-C6)alkyl group, and a halogen, provided that at least one group chosen among R1, R2, and R3 is not a hydrogen. Preferably R1 is not a hydrogen. In a preferred embodiment, R1 represents a —NR7R8 group with R7 and R8 being a hydrogen; R2 represents a hydrogen or a halogen, preferably a hydrogen; and R3 represents a hydrogen or a —NR7R8 group with R7 and R8 being a hydrogen, preferably a —NR7R8 group with R7 and R8 being a hydrogen.
In a more preferred embodiment, R1 represents a —NR7R8 group with R7 and R8 being a hydrogen; R2 represents a hydrogen; and R3 represents a —NR7R8 group with R7 and R8 being a hydrogen. According to this more preferred embodiment, R1 is —NH2, R2 is H, and R3 is —NH2.
According to the invention, R4 and R5 represent independently a hydrogen, a (C1-C6)alkyl group optionally substituted by a radical selected in the group consisting of a cycloalkyl, and a —NR9R10 group with R9 and R10 being independently a hydrogen or a (C1-C6)alkyl group, or R4 and R5 form together an azepanyl.
In a particular embodiment, R4 represents a hydrogen or a (C1-C6)alkyl group optionally substituted by a cycloalkyl or by a —NR9R10 group with R9 and R10 being a hydrogen. Preferably R4 represents a hydrogen, a propyl, an isopropyl, an isobutyl, a —CH2-cyclopropyl, an isopentyl, a butyl, a —(CH2)2—NH2, a —(CH2)3—NH2, a methyl, and an ethyl, preferably a propyl.
In a particular embodiment, R5 represents a (C1-C6)alkyl group, preferably a methyl group.
In a more particular embodiment, R4 represents a hydrogen or a (C1-C6)alkyl group optionally substituted by a cycloalkyl or by a —NR9R10 group with R9 and R10 being a hydrogen, preferably R4 represents a (C1-C6)alkyl group; and R5 represents a (C1-C6)alkyl group, preferably a methyl group.
In a preferred embodiment, R4 represents a propyl group and R5 represents a methyl group.
In a further particular embodiment, R4 and R5 form together an azepanyl.
According to the invention, R6 represents a hydrogen or a halogen. Preferably, R6 is a hydrogen.
According to the invention, n1, n2, and n3 are independently 0 or 1. In a particular embodiment, n1+n2+n3 is equal or superior to 1, preferably n1+n2+n3 is 1, 2, or 3.
According to the invention, n1 is 0 or 1. If n1 is 1, then X1 represents a —NHCO— group, an oxygen atom, a halogen, a —C≡C— group, or a —O—CH2— group. In a particular embodiment, n1 is 1 and X1 represents a —NHCO— group.
According to the invention, n2 is 0 or 1. If n2 is 1, then X2 represents a 5-12 membered ring optionally substituted by at least one radical selected in the group consisting of a (C1-C6)alkyl group optionally substituted by at least one halogen, a (C1-C6)alkoxy group optionally substituted by at least one halogen, a halogen, and a hydroxy.
In a particular embodiment, n2 is 1 and X2 represents a 5-12 membered ring selected in the group consisting of a phenyl, a pyrimidinyl, a thiophenyl, a pyridinyl, a triazolyl, and an indolyl, said 5-12 membered ring is optionally substituted by at least one radical selected in the group consisting of a methoxy, a trifluoromethoxy, a halogen, a hydroxy, a methyl, and a trifluromethyl.
According to the invention, n3 is 0 or 1. If n is 1, then X3 represents a radical selected in the group consisting of:
In a particular embodiment, n3 is 1 and X3 represents a radical selected in the group consisting of a (C1-C6)alkyl, a (C1-C6)alkoxy, a —C(O)—, a —SO2—, a —NH—SO2—, and a —NHCO—, said radical being substituted by a piperazinyl optionally substituted by a (C1-C6)alkyl, preferably a piperazinyl substituted by a methyl.
In a particular embodiment, n1 is 1. According to this particular embodiment, X1 represents a —NHCO— group, an oxygen atom, a halogen, a —C≡C— group, or a —O—CH2— group.
According to this particular embodiment, a compound of formula (I) or (I) is such that:
In a preferred embodiment, n1 is 1 and X1 represents a —NHCO— group.
According to this preferred embodiment, a compound of formula (I) or (I′) is such that:
In a further preferred embodiment, n1 is 1 and X1 represents an oxygen atom.
According to this preferred embodiment, a compound of formula (I′) is 4-(4-Aminopyrimidin-2-yl)-N-(2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)phenoxy)phenyl)thiazol-2-amine OR0143.
In a preferred embodiment, n1 is 1 and X1 represents a halogen, preferably a bromine. According to this preferred embodiment, a compound of formula (I′) is 2-(2-((5-Bromo-2-methylphenyl)(isobutyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0637-1; and 2-(2-((5-Bromo-2-methylphenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0652-1.
In a further preferred embodiment, n1 is 1 and X1 represents a —C≡C— group.
In a further preferred embodiment, n1 is 1 and X1 represents a —O—CH2— group.
According to this preferred embodiment, a compound of formula (I) is OR0232.
In a further particular embodiment, n1 is 0.
According to this particular embodiment, n2 is 1 and X2 represents a 5-12 membered ring selected in the group consisting of a phenyl, a pyrimidinyl, a thiophenyl, a pyridinyl, a triazolyl, and an indolyl, said 5-12 membered ring is optionally substituted by at least one radical selected in the group consisting of a methoxy, a trifluoromethoxy, a halogen, a hydroxy, a methyl, and a trifluromethyl.
According to this particular embodiment, n3 is 0 or 1, preferably 1 and X3 represents a radical selected in the group consisting of a (C1-C6)alkyl, a (C1-C6)alkoxy, a —C(O)—, a —SO2—, a —NH—SO2—, and a —NHCO—, said radical being substituted by at least one radical A as defined herein.
In a preferred embodiment, n3 is 1 and X3 represents a (C1-C6)alkyl substituted by at least one radical A as defined herein. Preferably said at least one radical A is a piperazinyl substituted by a methyl, a —N(CH3)2, and a —NHSO2CH3.
In a further preferred embodiment, n3 is 1 and X3 represents a —SO2— substituted by at least one radical A as defined herein. Preferably said at least one radical A is a piperazinyl optionally substituted by a methyl, an ethyl or a —CH2—COOH, a —NR11R12 group with Rn and R12 being independently a hydrogen or a methyl group, a —NH—(CH2)2—NH2, a morpholinyl, and a piperidinyl optionally substituted by a —NH2, more preferably a piperazinyl unsubstituted or substituted by a methyl, a —NR11R12 group with R11 and R12 being independently a hydrogen or a methyl group, or a morpholinyl.
In a further preferred embodiment, n3 is 1 and X3 represents a (C1-C6)alkoxy substituted by at least one radical A as defined herein, preferably a piperazinyl optionally substituted by a methyl or a —CO-piperazinyl optionally substituted by a methyl.
In a further preferred embodiment, n3 is 1 and X3 represents a —C(O)— substituted by at least one radical A as defined herein, preferably a piperazinyl optionally substituted by a methyl, a morpholinyl, or a methoxy.
In a further preferred embodiment, n3 is 1 and X3 represents a —NH—SO2— substituted by at least one radical A as defined herein, preferably a piperazinyl optionally substituted by a methyl.
In a further preferred embodiment, n3 is 1 and X3 represents a —NHCO— substituted by at least one radical A as defined herein, preferably a piperazinyl optionally substituted by a methyl.
In a further preferred embodiment, n3 is 1 and X3 represents a hydroxy.
In a further preferred embodiment, n3 is 1 and X3 represents a —NR15R16 group with R15 and R16 being independently a hydrogen or a (C1-C6)alkyl group, preferably a methyl.
A preferred compound of formula (I) or (I′) is such that:
A further preferred compound of formula (I) or (I′) is such that:
An even more preferred compound of formula (I) or (I) is such that:
A particular compound of formula (I) is selected in the group consisting of:
As illustrated by examples, the inventors have demonstrated the therapeutic interest of the compounds of the invention. Indeed, the inventors have shown that the compounds according to the invention are able to bind dCK and inhibit its activity, demonstrating thereby the therapeutic interest of such compounds in therapies, more particularly in cancer therapies. They have also demonstrated that such compounds, in combination with an antitumor drug, such as an inhibitor of the De Novo nucleotide biosynthesis pathway, significantly improved survival rate in a murine leukemia model.
Accordingly, the present invention relates to a compound of formula (I) or (I′) as defined herein, for use as a drug or a medicine. The present invention further relates to a pharmaceutical or veterinary composition comprising a compound according to the invention. Preferably, the pharmaceutical composition further comprises a pharmaceutically or veterinary acceptable carrier or excipient. The present invention relates to the use of a compound according to the invention as a drug or a medicine. The invention further relates to a method for treating a disease in a subject, wherein a therapeutically effective amount of a compound according to the invention, is administered to said subject in need thereof. The invention also relates to the use of a compound according to the invention, for the manufacture of a medicine. The invention also relates to a pharmaceutical composition comprising a compound according to the invention for use as a drug.
The present invention also concerns:
The term “cancer”, as used herein, refers to the presence of cells possessing characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and certain characteristic morphological features. The cancer may be a solid cancer, such as a solid tumor or a liquid cancer, such as a hematopoietic tumor. Examples of cancer include, for example, leukemia, lymphoma, blastoma, carcinoma, such as cholangiocarcinoma, and sarcoma. More particular examples of such cancers include chronic myeloid leukemia, acute lymphoblastic leukemia, such as T-cell acute lymphoblastic leukemia, Philadelphia chromosome positive acute lymphoblastic leukemia (Ph+ALL), squamous cell carcinoma, lung cancer, small-cell lung cancer, non-small cell lung cancer, glioma, gastrointestinal cancer, renal cancer, ovarian cancer, liver cancer, colorectal cancer, endometrial cancer, kidney cancer, prostate cancer, melanoma, skin cancer, thyroid cancer, neuroblastoma, osteosarcoma, pancreatic cancer, glioblastoma multiforme, cervical cancer, stomach cancer, bladder cancer, hepatoma, breast cancer, oesophagal cancer, colon cancer, head and neck cancer, brain cancer, gastric cancer, germ cell tumor, pediatric sarcoma, sinonasal natural killer, multiple myeloma, acute myelogenous leukemia (AML), chronic lymphocytic leukemia, mastocytosis and any symptom associated with mastocytosis. In a particular embodiment, the cancer is a liquid cancer, preferably acute lymphoblastic leukemia, more preferably T-cell acute lymphoblastic leukemia.
The term “therapy”, as used herein, refers to any type of treatment of cancer (i.e., antitumor therapy), including an adjuvant therapy and a neoadjuvant therapy. Therapy comprises radiotherapy and therapies, preferably systemic therapies such as hormone therapy, chemotherapy, immunotherapy and monoclonal antibody therapy.
The term “adjuvant therapy”, as used herein, refers to any type of treatment of cancer given as additional treatment, usually after surgical resection of the primary tumor, in a patient affected with a cancer that is at risk of metastasizing and/or likely to recur. The aim of such an adjuvant treatment is to improve the prognosis. Adjuvant therapies comprise radiotherapy and therapy, preferably systemic therapy, such as hormone therapy, chemotherapy, immunotherapy and monoclonal antibody therapy.
The term “hormone therapy” or “hormonal therapy” refers to a cancer treatment having for purpose to block, add or remove hormones. For instance, in breast cancer, the female hormones oestrogen and progesterone can promote the growth of some breast cancer cells. So, in these patients, hormone therapy is given to block oestrogen and a non-exhaustive list commonly used drugs includes: tamoxifen, toremifene, anastrozole, exemestane, letrozole, goserelin, leuprolide, megestrol acetate, and fluoxymesterone.
As used herein, the term “chemotherapeutic treatment” or “chemotherapy” refers to a cancer therapeutic treatment using chemical or biological substances, in particular using one or several antineoplastic agents.
The term “radiotherapeutic treatment” or “radiotherapy” is a term commonly used in the art to refer to multiple types of radiation therapy including internal and external radiation therapies or radioimmunotherapy, and the use of various types of radiations including X-rays, gamma rays, alpha particles, beta particles, photons, electrons, neutrons, radioisotopes, and other forms of ionizing radiations.
The term “therapeutical antibody” refers to any antibody having an anti-tumoral effect.
Preferably, the therapeutical antibody is a monoclonal antibody. Therapeutic antibodies are generally specific for surface antigens, e.g., membrane antigens. Most preferred therapeutic antibodies are specific for tumor antigens (e.g., molecules specifically expressed by tumor cells), such as CD20, CD52, ErbB2 (or HER2/Neu), CD33, CD22, CD25, MUC-1, CEA, KDR, aVb3, and the like. The therapeutical antibody includes, but is not limited to, antibodies such as trastuzumab (anti-HER2 antibody), rituximab (anti-CD20 antibody), alemtuzumab, gemtuzamab, cetuximab, pertuzumab, epratuzumab, basiliximab, daclizumab, labetuzumab, sevirumab, tuvurimab, palivizumab, infliximab, omalizumab, efalizumab, natalizumab, clenoliximab, and bevacizumab.
Hyperthermia is a medical treatment in which is exposed to high temperatures to damage and kill cancer cells or to make cancer cells more sensitive to the effects of radiation and certain anti-cancer drugs. There are many techniques, well-known by the one skilled in the art, by which heat may be delivered. Some of the most common involve the use of focused ultrasound (FUS or HIFU), infrared sauna, microwave heating, induction heating, magnetic hyperthermia, infusion of warmed liquids, or direct application of heat such as through sitting in a hot room or wrapping a patient in hot blankets.
The administration route can be topical, transdermal, oral, rectal, sublingual, intranasal, intrathecal, intratumor or parenteral (including subcutaneous, intramuscular, intravenous and/or intradermal). Preferably, the administration route is parental, oral or topical. The pharmaceutical composition is adapted for one or several of the above-mentioned routes. The pharmaceutical composition, kit, product or combined preparation is preferably administered by injection or by intravenous infusion or suitable sterile solutions, or in the form of liquid or solid doses via the alimentary canal.
The pharmaceutical composition can be formulated as solutions in pharmaceutically compatible solvents or as emulsions, suspensions or dispersions in suitable pharmaceutical solvents or vehicles, or as pills, tablets or capsules that contain solid vehicles in a way known in the art. Formulations of the present invention suitable for oral administration may be in the form of discrete units as capsules, sachets, tablets or lozenges, each containing a predetermined amount of the active ingredient; in the form of a powder or granules; in the form of a solution or a suspension in an aqueous liquid or non-aqueous liquid; or in the form of an oil-in-water emulsion or a water-in-oil emulsion. Formulations for rectal administration may be in the form of a suppository incorporating the active ingredient and carrier such as cocoa butter, or in the form of an enema. Formulations suitable for parenteral administration conveniently comprise a sterile oily or aqueous preparation of the active ingredient which is preferably isotonic with the blood of the recipient. Every such formulation can also contain other pharmaceutically compatible and nontoxic auxiliary agents, such as, e.g. stabilizers, antioxidants, binders, dyes, emulsifiers or flavoring substances. The formulations of the present invention comprise an active ingredient in association with a pharmaceutically acceptable carrier therefore and optionally other therapeutic ingredients. The carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulations and not deleterious to the recipient thereof. The pharmaceutical compositions are advantageously applied by injection or intravenous infusion of suitable sterile solutions or as oral dosage by the digestive tract. Methods for the safe and effective administration of most of these chemotherapeutic agents are known to those skilled in the art. In addition, their administration is described in the standard literature.
Pharmaceutical compositions according to the invention may be formulated to release the active drug substantially immediately upon administration or at any predetermined time or time period after administration. Preferably, the treatment with the compound according to the invention or the pharmaceutical composition according to the invention starts no longer than a month, preferably no longer than a week, after the diagnosis of the disease. In a most preferred embodiment, the treatment starts the day of the diagnosis.
The compound according to the invention or the pharmaceutical composition according to the invention may be administered as a single dose or in multiple doses.
Preferably, the treatment is administered regularly, preferably between every day and every month, more preferably between every day and every two weeks, more preferably between every day and every week, even more preferably the treatment is administered every day. In a particular embodiment, the treatment is administered several times a day, preferably 2 or 3 times a day, even more preferably 3 times a day.
The duration of treatment with the compound according to the invention or the pharmaceutical composition according to the invention is preferably comprised between 1 day and 50 weeks, more preferably between 1 day and 30 weeks, still more preferably between 1 day and 15 weeks, even more preferably between 1 day and 10 weeks. In a particular embodiment, the duration of the treatment is of about 1 week. Alternatively, the treatment may last as long as the disease persists.
The amount of compound according to the invention or of pharmaceutical composition according to the invention to be administered has to be determined by standard procedure well known by those of ordinary skills in the art. Physiological data of the patient (e.g. age, size, and weight) and the routes of administration have to be taken into account to determine the appropriate dosage, so as a therapeutically effective amount will be administered to the patient.
In a preferred embodiment, the total compound dose for each administration of the compound according to the invention or of the pharmaceutical composition according to the invention is comprised between 0.00001 and 1 g, preferably between 0.01 and 10 mg. The form of the pharmaceutical compositions, the route of administration and the dose of administration of the compound according to the invention, or the pharmaceutical composition according to the invention can be adjusted by the man skilled in the art according to the type and severity of the disease, and to the patient, in particular its age, weight, sex, and general physical condition.
In an embodiment, the compound of the invention can be used in combination with another antitumor drug or antineoplastic agent. The additional antitumor drug can be selected in the non-exhaustive list of antitumor agents consisting of an inhibitor of topoisomerases I or II, an anti-mitotic agent, a DNA alkylating agent, an agent causing crosslinking of DNA, an anti-metabolic agent, a targeted agent such as a kinase inhibitor, a histone deacetylase inhibitor and an anti-EGFR agent and/or a therapeutical antibody designed to mediate cytotoxicity against the cancer cells or to modulate one of their key biological functions.
Antimitotic agents include, but are not limited to, paclitaxel, docetaxel and analogs such as larotaxel (also called XRP9881; Sanofi-Aventis), XRP6258 (Sanofi-Aventis), BMS-184476 (Bristol-Meyer-Squibb), BMS-188797 (Bristol-Meyer-Squibb), BMS-275183 (Bristol-Meyer-Squibb), ortataxel (also called IDN 5109, BAY 59-8862 or SB-T-101131; Bristol-Meyer-Squibb), RPR 109881A (Bristol-Meyer-Squibb), RPR 116258 (Bristol-Meyer-Squibb), NBT-287 (TAPESTRY), PG-paclitaxel (also called CT-2103, PPX, paclitaxel poliglumex, paclitaxel polyglutamate or Xyotax™), ABRAXANE® (also called Nab-paclitaxel; ABRAXIS BIOSCIENCE), tesetaxel (also called DJ-927), IDN 5390 (INDENA), taxoprexin (also called docosahexanoic acid-paclitaxel; PROTARGA), DHA-paclitaxel (also called Taxoprexin®), and MAC-321 (WYETH). Preferably, antimitotic agents are docetaxel, paclitaxel, and is more preferably docetaxel.
Inhibitors of topoisomerases I and/or II include, but are not limited to etoposide, topotecan, camptothecin, irinotecan, amsacrine, intoplicin, anthracyclines such as doxorubicin, epirubicin, daunorubicin, idarubicin and mitoxantrone. Inhibitors of topoisomerase I and II include, but are not limited to intoplicin.
The additional antitumor agent can be alkylating agents including, without limitation, nitrogen mustards, ethylenimine derivatives, alkyl sulfonates, nitrosoureas, metal salts and triazenes. Non-exhaustive examples thereof include uracil mustard, chlormethine, cyclophosphamide (CYTOXAN®), ifosfamide, melphalan, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, cisplatin, carboplatin, fotemustine, oxaliplatin, thiotepa, streptozocin, dacarbazine, and temozolomide. In a preferred embodiment, the DNA alkylating agent is preferably cisplatin, carboplatin, temozolomide, fotemustine or dacarbazine.
Anti-metabolic agents block the enzymes responsible for nucleic acid synthesis or become incorporated into DNA, which produces an incorrect genetic code and leads to apoptosis. Non-exhaustive examples thereof include, without limitation, folic acid antagonists, pyrimidine analogs, purine analogs and adenosine deaminase inhibitors, and more particularly methotrexate, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, pentostatine, 5-fluorouracil, gemcitabine and capecitabine. In a preferred embodiment, such an agent is gemcitabine.
The additional anti-tumor agent can also be a targeted agent, in particular a kinase inhibitor. The kinase may be selected from the group consisting of intracellular tyrosine or serine/threonine kinases, receptors tyrosine or serine/threonine kinase. The kinase could be selected among EGFR family, ALK, B-Raf, MEK, and mTOR. For instance, the agents may have ability to inhibit angiogenesis based on the inhibitory activities on VEGFR and PDGFR kinases. In particular, the targeted agent can be selected among the multiple kinase inhibitor drugs which are already approved: Gleevec®, which inhibits Bcr-Abl and c-Kit, and Iressa® and Tarceva®, which both inhibit EGFR, sorafenib (Nexavar®, BAY 43-9006) which inhibits Raf, dasatinib (BMS-354825) and nilotinib (AMN-107, Tasigna®) which also inhibits Bcr-Abl, lapatinib which also inhibits EGFR, temsirolimus (Torisel®, CCI-779) which targets the mTOR pathway, sunitinib (Student, SU11248) which inhibits several targets including VEGFR as well as specific antibodies inactivating kinase receptors: Herceptin® and Avastin®. The anti-EGFR agent can be selected among gefitinib, erlotinib, lapatinib, vandetanib, afatinib, osimertinib, neratinib, dacomitinib, brigatinib, canertinib, naquotinib, nazartinib, pelitinib, rociletinib, icotinib, AZD3759, AZ5104 (CAS No 1421373-98-9), poziotinib, WZ4002, preferably is erlotinib or cetuximab. The ALK inhibitor can be selected among crizotinib, entrectinib, ceritinib, alectinib, brigatinib, lorlatinib, TSR-011, CEP-37440, and ensartinib. The B-Raf inhibitor can be selected among vemurafenib, dabrafenib, regorafenib, and PLX4720. The MEK inhibitor can be selected among cobimetinib, trametinib, binimetinib, selumetinib, PD-325901, CI-1040, PD035901, U0126, TAK-733.
The additional drug can also be a checkpoint inhibitor, for instance an antibody targeting PD-1, PD-L1, CTLA-4 and the like.
In a preferred embodiment, the compound of the invention can be used in combination with an inhibitor of the De Novo nucleotide biosynthesis pathway. In De Novo (from scratch) pathways, the nucleotide bases are assembled from simpler compounds. The framework for a pyrimidine base is assembled first and then attached to ribose. In contrast, the framework for a purine base is synthesized piece by piece directly onto a ribose-based structure. De novo pathways synthesize pyrimidines and purine nucleotides from amino acids, carbon dioxide, folate derivatives, and phosphoribosyl pyrophosphate (PRPP).
As used herein the terms “De Novo nucleotide biosynthesis pathway inhibitor”, “inhibitor of the De Novo nucleotide biosynthesis pathway” or simply “De Novo pathway inhibitor” or “inhibitor of the De Novo pathway” refers to any agent (e.g., compound, antibody, protein, or nucleic acid) capable of reducing the level of a protein component of the De Novo nucleotide biosynthesis pathway, an mRNA of a protein component of the De Novo nucleotide biosynthesis pathway, or the activity of a component of the De novo nucleotide biosynthesis pathway, relative to a control (e.g., comparison of level in the absence of the De Novo pathway inhibitor). Preferably, the De Novo Pathway inhibitor is a compound (e.g., molecule). The De Novo pathway inhibitor may reduce the level of production of dCTP, and/or dATP and/or dGTP and dTTP compared to control (e.g., absence of the De Novo Pathway inhibitor). Non-limiting examples of De Novo Pathway inhibitor are listed below:
In a more preferred embodiment, the inhibitor of the De Novo pathway is a ribonucleotide reductase (RNR) inhibitor. As used herein the terms “inhibitor of ribonucleotide reductase” or “ribonucleotide reductase inhibitor refer to an agent (e.g., chemical compound, antibody, protein, or nucleic acid) capable of reducing the level of RNR protein, RNR mRNA, or RNR activity, relative to a control (e.g., comparison of level in the absence of the RNR inhibitor). Preferably, the RNR inhibitor is a chemical compound (e.g., a molecule). The RNR inhibitor may reduce the level of activity of RNR. The RNR inhibitor may reduce the level of activity of RNR when the RNR inhibitor binds RNR. Non-limiting examples of RNR inhibitors are listed below:
In an even more preferred embodiment, the ribonucleotide reductase inhibitor is thymidine.
Further aspects and advantages of the invention will be disclosed in the following experimental section.
General Methods. Commercially available reagents and solvents were used without further additional purification. Thin layer chromatography (TLC) was performed on precoated aluminum sheets of silica (60 F254 nm, Merck) and visualized using short-wave UV light. Reaction monitoring and purity of compounds were recorded by using analytical Agilent Infinity high performance liquid chromatography with DAD at 254 nm column Agilent Poroshell 120 EC-C18 2.7 μm (4.6×50 mm), mobile phase (A: 0.1% FA H2O, B: 0.1% FA MeCN), method (A) flow rate 0.3 mL/min, time/% B 0/10, 4/90, 7/90, 9/10, 10/10; method (B) flow rate 0.5 mL/min, time/% B 0/10, 4/90, 7/90, 9/10, 13/10); mobile phase (A: 0.1% TFA H2O, B: 0.1% TFA MeCN), method (C) flow rate 1 mL/min, time/% B 0/10, 5/100, 8/100; column Thermo Scientific Hypersil Gold 12 μm (4.6×250 mm), mobile phase (A: 0.1% TFA H2O, B: 0.1% TFA MeCN), method (D) flow rate 2 mL/min, time/% B 0/10, 6/100, 11/100. Column chromatography was performed on a Reveleris purification system using Reveleris Flash silica cartridges or C18 40 μM cartridges. Petroleum refers to the fraction with distillation range 40-65° C. 1H and 13C NMR spectra were recorded by using a Bruker AC 400, AC300 or AC250 spectrometer. Chemical shifts, (δ) are reported in ppm and coupling values (J) in hertz. Abbreviations for peaks are, br: broad, s: singlet, d: doublet, t: triplet, q: quadruplet, quint: quintuplet, sext: sextuplet, sept: septuplet and m: multiplet). The spectra recorded are consistent with the proposed structures. Low-resolution mass spectra were obtained with Agilent SQ G6120B mass spectrometer in positive and negative electrospray mode.
General methods for preparing compounds of the invention are illustrated by Schemes 1-7.
N-(3-((4-(4-aminopyrimidin-2-yl)thiazol-2-yl)amino)-4-aryl)benzamide derivatives were prepared by convergent synthesis, from commercially available 4-amino-2-chloropyrimidine and appropriate nitroaniline (Scheme 1). The 4-amino-2-chloropyrimidine was protected using Boc2O to afford corresponding bis-carbamate. A Stille cross-coupling reaction with tributyl(1-ethoxyvinyl)tin gives the enol ether which was then turned into the corresponding α-bromoketone using N-bromosuccinimide. Starting from appropriate nitroaniline, condensation with acetylisothiocyanate followed by saponification gave the corresponding thiourea. The α-bromoketone was engaged in the Hantzsch thiazole synthesis with the thiourea leading to the corresponding thiazole. Optionally R4 as (C1-C6)alkyl group can be introduced before reduction of the nitro group followed by peptide coupling with appropriate carboxylic acid affording expected amides. Finally, deprotection with TFA led to corresponding N-(3-((4-(4-aminopyrimidin-2-yl)thiazol-2-yl)amino)-4-aryl)benzamides.
General Procedure for the Synthesis of N-(3-((4-(4-aminopyrimidin-2-yl)thiazol-2-yl)amino)-4-aryl)benzamide Derivatives. A solution of N-(3-((4-(4-(di-tert-butoxycarbonylamino)pyrimidin-2-yl)thiazol-2-yl)amino)-4-aryl)benzamide derivative (0.1 mmol) in a mixture of dichloromethane-trifluoroacetic acid (3:1, 4 mL) was stirred at room temperature for 2 hrs. The solvent was distillated off under reduced pressure and the residue was purified by flash chromatography to afford the corresponding N-(3-((4-(4-aminopyrimidin-2-yl)thiazol-2-yl)amino)-4-aryl)benzamide.
N-(3-((4-(4-Aminopyrimidin-2-yl)thiazol-2-yl)amino)-4-methylphenyl)-4-((4-methylpiperazin-1-yl)methyl)benzamide dCKi-1 (78%) as a white powder. 1H NMR (400 MHz, MeOD) δ 8.40 (d, J=1.7 Hz, 1H), 8.13 (d, J=6.0 Hz, 1H), 7.96 (d, J=8.2 Hz, 2H), 7.66 (dd, J=8.2, 1.7 Hz, 1H), 7.60 (s, 1H), 7.49 (d, J=8.2 Hz, 2H), 7.21 (d, J=8.2 Hz, 1H), 6.44 (d, J=6.0 Hz, 1H), 3.61 (brs, 2H), 2.53 (brs, 8H), 2.32 (s, 3H), 2.30 (s, 3H); 13C NMR (100 MHz, MeOD) δ 168.4, 167.5, 165.5, 161.2, 155.8, 151.1, 142.9, 140.7, 138.8, 135.4, 131.9, 130.6, 128.7, 125.5, 117.0, 113.9, 112.7, 104.6, 63.3, 55.7, 53.5, 45.9, 17.6; LCMS C27H30FN8OS method (D) Rt=3.579 min, ESI+ m/z=515.2 (M+H).
N-(1-(4-(4-Aminopyrimidin-2-yl)thiazol-2-yl)-2,3,4,5-tetrahydro-1H-benzo[b]azepin-8-yl)-4-((4-methylpiperazin-1-yl)methyl)benzamide OR0289 (66%) as a white solid. Rf=0.40 (DCM-MeOH—NH4OH, 85:13.5:1.5); 1H NMR (400 MHz, MeOD) δ 8.08 (d, J=6.0 Hz, 1H), 7.89 (d, J=8.2 Hz, 2H), 7.83 (d, J=2.0 Hz, 1H), 7.62 (dd, J=8.2, 2.1 Hz, 1H), 7.44 (d, J=8.2 Hz, 2H), 7.37 (s, 1H), 7.30 (d, J=8.3 Hz, 1H), 6.41 (d, J=6.0 Hz, 1H), 3.95 (m, 2H), 3.58 (s, 2H), 2.78-2.67 (m, 2H), 2.57 (s, 8H), 2.34 (s, 3H), 1.93 (m, 2H), 1.69 (m, 2H); 13C NMR (100 MHz, MeOD) δ 170.78, 168.40, 165.47, 161.24, 155.30, 151.56, 146.10, 142.94, 139.56, 138.08, 135.09, 132.21, 130.50, 128.78, 121.77, 121.45, 112.18, 104.50, 63.08, 55.57, 53.20, 52.20, 45.63, 34.87, 29.52, 27.58; LC method (C) Rt=2.822 min.
N-(3-((4-(4-Aminopyrimidin-2-yl)thiazol-2-yl)(propyl)amino)-4-methylphenyl)-4-((4-methylpiperazin-1-yl)methyl)benzamide OR0105 (86%) as a white solid. Rf=0.18 (DCM-MeOH—NH4OH, 90:9:1). 1H NMR (400 MHz, MeOD) δ 8.14 (d, J=6.0 Hz, 1H), 7.93 (d, J=8.3 Hz, 2H), 7.78 (d, J=2.0 Hz, 1H), 7.72 (dd, J=8.3, 2.2 Hz, 1H), 7.51 (d, J=8.2 Hz, 2H), 7.41 (d, J=8.4 Hz, 1H), 7.38 (s, 1H), 6.45 (d, J=6.0 Hz, 1H), 4.01 (brs, 2H), 3.64 (s, 2H), 2.57 (brs, 8H), 2.34 (s, 3H), 2.26 (s, 3H), 1.86-1.69 (m, 2H), 1.02 (t, J=7.4 Hz, 3H); LC method (C) Rt=3.967 min.
N-(3-((4-(4-Aminopyrimidin-2-yl)thiazol-2-yl)(propyl)amino)-4-methylphenyl)-2-((4-methylpiperazin-1-yl)methyl)pyrimidine-5-carboxamide OR0125 (81%) as a light orange powder. Rf=0.23 (DCM-MeOH—NH4OH, 80:20:0.2); 1H NMR (400 MHz, MeOD) δ 9.33 (s, 2H), 8.54 (d, J=1.9 Hz, 1H), 8.14 (d, J=6.0 Hz, 1H), 7.70 (dd, J=8.2, 1.9 Hz, 1H), 7.63 (s, 1H), 7.21 (d, J=8.2 Hz, 1H), 6.45 (d, J=6.0 Hz, 1H), 3.91 (s, 2H), 2.82-2.46 (m, 8H), 2.33 (s, 6H); LC method (D) Rt=3.967 min.
N-(3-((4-(4-Aminopyrimidin-2-yl)thiazol-2-yl)amino)-4-methylphenyl)-4-(2-(4-methylpiperazin-1-yl)ethyl)benzamide OR0146 (49%) as orange crystals. Rf=0.55 (DCM-MeOH—NH4OH, 80:18:2). 1H NMR (400 MHz, MeOD) δ 8.42 (d, J=2.0 Hz, 1H), 8.10 (d, J=6.0 Hz, 1H), 7.88 (d, J=8.3 Hz, 2H), 7.61 (dd, J=8.5, 2.0 Hz, 1H), 7.59 (s, 1H), 7.32 (d, J=8.3 Hz, 2H), 7.16 (d, J=8.5 Hz, 1H), 6.41 (d, J=6.0 Hz, 1H), 2.88-2.81 (m, 2H), 2.65-2.59 (m, 2H), 2.59 (s, 8H), 2.32 (s, 3H), 2.29 (s, 3H); 13C NMR (100 MHz, MeOD) δ 168.39, 167.34, 165.46, 160.96, 155.39, 150.83, 145.51, 140.60, 138.79, 134.16, 131.85, 129.92, 128.86, 125.26, 116.90, 113.78, 112.99, 104.63, 60.58, 55.45, 53.28, 45.69, 33.83, 17.63; LC method (D) Rt=3.592 min.
N-(3-((4-(4-Amino-5-fluoropyrimidin-2-yl)thiazol-2-yl)amino)-4-methylphenyl)-4-((4-methylpiperazin-1-yl)methyl)benzamide OR0239 (57%) as a white powder. 1H NMR (400 MHz, MeOD) δ 8.33 (d, J=1.8 Hz, 1H), 8.10 (d, J=3.6 Hz, 1H), 7.95 (d, J=8.2 Hz, 2H), 7.64 (dd, J=8.3, 1.8 Hz, 1H), 7.54 (s, 1H), 7.49 (d, J=8.2 Hz, 2H), 7.21 (d, J=8.3 Hz, 1H), 3.62 (brs, 2H), 2.52 (brs, 8H), 2.32 (s, 3H), 2.29 (s, 3H); LC method (D) Rt=3.548 min N-(3-((4-(4-Aminopyrimidin-2-yl)thiazol-2-yl)amino)-4-methylphenyl)-4-((dimethylamino)methyl)benzamide OR0155. (77%) as a white solid. Rf=0.20 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ8.39 (s, 1H), 8.11 (s, 1H), 7.96 (d, J=8.1 Hz, 2H), 7.62 (m, 1H), 7.60 (s, 1H), 7.47 (d, J=8.21 Hz, 2H), 7.20 (d, J=8.01 Hz, 1H), 6.43 (d, J=6.1 Hz, 1H), 3.60 (s, 2H), 2.31 (s, 3H), 2.30 (s, 6H); 13C NMR (100 MHz, MeOD) S 168.32, 167.53, 165.52, 161.13, 155.60, 152.54, 142.42, 140.58, 138.80, 135.63, 131.92, 130.83, 128.81, 125.52, 117.08, 113.97, 112.80, 104.67, 64.30, 45.19, 17.59; LC method (D) Rt=3.703 min.
N-(3-((4-(4-Aminopyrimidin-2-yl)thiazol-2-yl)amino)-4-methylphenyl)-4-((6-methyl-2,6-diazaspiro[3.3]heptan-2-yl)methyl)benzamide OR0156 (10% over 2 steps) as a white solid. Rf=0.18 (DCM-MeOH—NH4OH, 80:20:2). 1H NMR (400 MHz, MeOD) δ 8.42 (d, J=2.0 Hz, 1H), 8.13 (d, J=6.0 Hz, 1H), 7.96 (d, J=8.1 Hz, 2H), 7.63 (s, 1H), 7.52 (dd, J=8.2, 2.0 Hz, 1H), 7.43 (d, J=8.1 Hz, 2H), 7.21 (d, J=8.3 Hz, 1H), 6.46 (d, J=6.1 Hz, 1H), 3.98 (s, 4H), 3.72 (s, 2H), 3.49 (s, 4H), 2.69 (s, 3H), 2.33 (s, 3H).
N-(3-((4-(4-Aminopyrimidin-2-yl)thiazol-2-yl)amino)-4-methylphenyl)-5-((4-methylpiperazin-1-yl)methyl)thiophene-2-carboxamide OR0241 (83%) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 10.17 (s, 1H), 9.37 (brs, 1H), 8.08 (d, J=5.8 Hz, 1H), 7.98 (d, J=1.8 Hz, 1H), 7.83 (d, J=3.8 Hz, 1H), 7.47 (s, 1H), 7.45 (dd, J=8.4, 1.8 Hz, 1H), 7.20 (d, J=8.4 Hz, 1H), 7.02 (d, J=3.8 Hz, 1H), 6.79 (brs, 2H), 6.31 (d, J=5.8 Hz, 1H), 3.68 (s, 2H), 2.42 (brs, 4H), 2.33 (brs, 4H), 2.23 (s, 3H), 2.15 (s, 3H); LC method (D) Rt=3.725 min.
General Procedure for the Synthesis of di-tert-butyl (2-(2-((3-benzamidoaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate Derivatives. To a solution of di-tert-butyl (2-(2-((3-aminoaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate derivative (0.37 mmol) and appropriate acid (0.56 mmol) in dry dimethyformamide (6 mL) were successively added 0-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (170 mg, 0.44 mmol), in one portion, and diisopropylethylamine (330 μL, 1.85 mmol). The resulting mixture was stirred overnight at room temperature and then concentrated under reduced pressure. The residue was diluted with EtOAc (100 mL) and successively washed twice with water (20 mL) and saturated aqueous Na2CO3 (20 mL). The organic layer was dried over Na2SO4, and the solvent was distillated off under reduced pressure. The residue was purified by flash chromatography to afford the corresponding di-tert-butyl (2-(2-((3-benzamidoaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate.
Di-tert-butyl (2-(2-((2-methyl-5-(4-(2-(4-methylpiperazin-1-yl)methyl)benzamido)phenyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate dCKi-1-1 (38%) as a white powder. 1H NMR (300 MHz, CDCl3) 6.68 (d, J=5.7, 1H), 8.19 (s, 1H), 7.93 (brs, 1H), 7.84 (d, J=8.0, 2H), 7.55 (s, 1H), 7.54 (d, J=5.6, 1H), 7.45-7.41 (m, 3H), 7.22-7.18 (m, 2H), 3.57 (s, 2H), 2.51 (brs, 8H), 2.33 (s, 3H), 2.29 (s, 3-H), 1.55 (s, 18H); 13C NMR (75 MHz, CDCl3) δ 165.7, 165.5, 159.9, 158.6, 150.3, 149.7, 142.8, 138.9, 137.4, 133.8, 131.6, 129.5, 127.2, 124.2, 115.3, 112.5, 110.9, 110.6, 84.4, 77.4, 62.6, 52.2, 53.0, 46.0, 28.0, 17.6; LCMS C37H46N8O5S method (D) Rt=4.908 min, ESI+ m/z=715.3 (M+H).
Di-tert-butyl (2-(2-(8-(4-((4-methylpiperazin-1-yl)methyl)benzamido)-2,3,4,5-tetrahydro-1H-benzo[b]azepin-1-yl)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0289-1 (45%) as a white foamy solid. Rf=0.30 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) S 8.74 (d, J=5.7 Hz, 1H), 7.91 (d, J=8.2 Hz, 2H), 7.85 (d, J=2.0 Hz, 1H), 7.64 (dd, J=8.3, 2.1 Hz, 1H), 7.56 (d, J=5.7 Hz, 1H), 7.49 (d, J=8.3 Hz, 2H), 7.46 (s, 1H), 7.35 (d, J=8.3 Hz, 1H), 4.05 (brs, 2H), 3.67 (s, 2H), 2.95-2.82 (brs, 4H), 2.69-2.50 (brs, 4H), 2.75 (m, 2H), 2.59 (s, 3H), 1.98 (s, 2H), 1.74 (m, 2H), 1.54 (s, 18H); 13C NMR (100 MHz, MeOD) δ 171.20, 168.46, 161.55, 160.25, 159.48, 151.57, 151.18, 146.12, 142.77, 139.61, 138.22, 135.24, 132.29, 130.45, 128.82, 121.86, 121.46, 113.68, 113.03, 85.64, 62.68, 55.31, 52.29, 52.19, 44.76, 34.89, 29.57, 28.07, 27.62; LC method (C) Rt=4.143 min.
Di-tert-butyl (2-(2-((2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)benzamido)phenyl)(propyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0105-1 (55%) as a white powder used in the next step without further purification.
Di-tert-butyl (2-(2-((2-methyl-5-(2-((4-methylpiperazin-1-yl)methyl)pyrimidine-5-carboxamido)phenyl)(propyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0125-1 (26%) as an orange powder. Rf=0.42 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 9.38 (s, 2H), 8.69 (d, J=5.7 Hz, 1H), 8.34 (d, J=1.3 Hz, 1H), 7.58 (dd, J=8.3, 1.3 Hz, 1H), 7.52 (d, J=5.7 Hz, 1H), 7.49 (s, 1H), 7.16 (d, J=8.3 Hz, 1H), 3.91 (s, 2H), 3.09 (brs, 8H), 2.31 (s, 6H), 1.53 (s, 18H).
Di-tert-butyl (2-(2-((2-methyl-5-(4-(2-(4-methylpiperazin-1-yl)ethyl)benzamido)phenyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0146-1 (28%) as a white foamy solid. Rf=0.36 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 8.73 (d, J=5.6 Hz, 1H), 8.36 (d, J=1.4 Hz, 1H), 7.92 (d, J=8.1 Hz, 2H), 7.64 (s, 1H), 7.61 (dd, J=8.3, 1.5 Hz, 1H), 7.59 (d, J=5.5 Hz, 1H), 7.39 (d, J=8.1 Hz, 2H), 7.19 (d, J=8.3 Hz, 1H), 3.09 (brs, 4H), 2.91 (m, 2H), 2.85 (brs, 4H), 2.80 (m, 2H), 2.72 (s, 3H), 2.30 (s, 3H), 1.53 (s, 18H); 13C NMR (100 MHz, MeOD) δ 168.40, 168.16, 161.12, 160.25, 159.55, 151.40, 150.24, 145.04, 140.44, 138.76, 134.14, 131.98, 130.06, 128.93, 125.90, 117.41, 114.48, 114.29, 113.00, 85.75, 59.58, 54.71, 51.67, 44.15, 33.59, 28.09, 17.64; LC method (D) Rt=4.960 min.
Di-tert-butyl (5-fluoro-2-(2-((2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)benzamido)phenyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0239-1 (31%) as a light brown powder. 1H NMR (400 MHz, CDCl3) δ 8.65 (d, J=1.3 Hz, 1H), 8.19 (d, J=1.8 Hz, 1H), 7.84 (d, J=8.2 Hz, 2H), 7.78 (dd, J=8.2, 1.3 Hz, 1H), 7.61 (s, 1H), 7.45 (d, J=8.2 Hz, 2H), 7.21 (d, J=8.2 Hz, 1H), 3.57 (s, 2H), 2.48 (brs, 8H), 2.31 (s, 3H), 2.29 (s, 3H), 1.45 (s, 18H).
Di-tert-butyl (2-(2-((2-methyl-5-(5-((4-methylpiperazin-1-yl)methyl)thiophene-2-carboxamido)phenyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0241-1 (31%) as a yellow powder. Rf=0.53 (DCM-MeOH—NH4OH, 95:5:0.5); 1H NMR (400 MHz, CDCl3) δ 8.67 (d, J=5.7 Hz, 1H), 8.18 (d, J=1.8 Hz, 1H), 8.07 (brs, 1H), 7.60-7.50 (m, 3H), 7.39 (brs, 1H), 7.26-7.18 (m, 2H), 6.90 (d, J=3.6 Hz, 1H), 3.74 (s, 2H), 2.67 (brs, 8H), 2.44 (s, 3H), 2.27 (s, 3H), 1.56 (s, 18H).
Di-tert-butyl (2-(2-((5-(4-((dimethylamino)methyl)benzamido)-2-methylphenyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0155-1.
Under argon, at 0° C., to a solution of di-tert-butyl (2-(2-((5-amino-2-methylphenyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate dCKi-1-2 (195 mg, 0.391 mmol), in a mixture of tetrahydrofuran-dimethylformamide (4:1, 5 mL) was successively added N,N-diisopropylethylamine (DIPEA) (400 μL, 2.30 mmol) and a freshly prepared suspension of 4-((dimethylamino)methyl)benzoyl chloride hydrochloride (108 mg, 0.463 mmol) in tetrahydrofuran (4 mL). After addition, the resulting mixture was stirred overnight at room temperature. The solvent was distillated off under reduced pressure and the residue was taken up into saturated Na2CO3 aqueous solution (20 mL) and extracted with EtOAc (2×50 mL). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The residue was purified by flash chromatography, gradient (DCM-MeOH—NH4OH, 90:10:1) to afford 4-((dimethylamino)methyl)-N-(3-((4-(4-(di-tertbutoxycarbonylamino)pyrimidin-2-yl)thiazol-2-yl)amino)-4-methylphenyl)benzamide OR0155-1 (75 mg, 29%) as a white solid. Rf=0.10 (DCM-MeOH—NH4OH, 95:5:1). 1H NMR (400 MHz, CDCl3) δ 9.09 (s, 1H), 8.54 (s, 1H), 8.17 (s, 1H), 7.88 (d, J=6.8 Hz, 2H), 7.53-7.34 (m, 5H), 7.09 (d, J=7.6 Hz, 1H), 3.70 (s, 2H), 2.37 (s, 6H), 2.20 (s, 3H), 1.52 (s, 18H). 13C NMR (100 MHz, CDCl3) δ 166.19, 165.81, 159.74, 158.44, 158.32, 150.21, 149.51, 138.81, 137.52, 134.84, 131.38, 130.06, 127.88, 124.86, 116.21, 114.15, 112.39, 111.95, 110.69, 84.44, 62.65, 44.26, 27.90, 17.52; LC method (D) Rt=5.229 min.
General Procedure for the Synthesis of di-tert-butyl (2-(2-((3-aminoaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate derivatives. To a solution of di-tert-butyl (2-(2-((3-nitroaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate derivative (0.5 mmol) in a mixture of tetrahydrofuran-ethanol (10:1, 22 mL) was added tin (II) chloride dihydrate (1.14 g, 5.06 mmol). The reaction mixture was stirred overnight at room temperature until the reaction was complete as indicated by TLC or LCMS monitoring. The reaction was quenched with a saturated aqueous Na2CO3 solution (20 mL) and stirred for further 15 min. After dilution with EtOAc (70 mL) and filtration through a short pad of celite, the aqueous layer was extracted twice with EtOAc (70 mL). The combined organic layers were dried over Na2SO4 and the solvent was distillated off under reduced pressure. The residue was purified by flash chromatography to afford the corresponding di-tert-butyl (2-(2-((3-aminoaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate.
Di-tert-butyl (2-(2-((5-amino-2-methylphenyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate dCKi-1-2 (77%) as an orange foamy solid. Rf=0.40 (DCM-MeOH, 95:5); 1H NMR (400 MHz, CDCl3) δ 8.65 (d, J=5.7 Hz, 1H), 7.51 (d, J=5.7 Hz, 1H), 7.48 (s, 1H), 7.00 (d, J=8.1 Hz, 1H), 6.89 (d, J=2.3 Hz, 1H), 6.43 (dd, J=8.0, 2.3 Hz, 1H), 3.67 (s, 2H), 2.18 (s, 3H), 1.55 (s, 18H); 13C NMR (75 MHz, CDCl3) δ 166.6, 158.6, 150.3, 149.9, 145.8, 139.2, 132.0, 119.3, 111.8, 111.7, 111.0, 107.1, 84.4, 77.4, 28.0, 17.1; LCMS C24H30N6O4S method (D) Rt=5.154 min, ESI+ m/z=499.2 (M+H).
Di-tert-butyl (2-(2-(8-amino-2,3,4,5-tetrahydro-1H-benzo[b]azepin-1-yl)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0289-2 (71%) as a yellow foamy solid. Rf=0.20 (DCM-MeOH, 95:5); 1H NMR (400 MHz, CDCl3) δ 8.70 (d, J=5.7 Hz, 1H), 7.47 (d, J=5.7 Hz, 1H), 7.37 (s, 1H), 7.04 (d, J=8.1 Hz, 1H), 6.72 (d, J=2.1 Hz, 1H), 6.57 (dd, J=8.0, 2.3 Hz, 1H), 3.97 (brs, 2H), 3.45 (brs, 2H), 2.70-2.55 (m, 2H), 1.90 (m, 2H), 1.63 (s, 2H), 1.53 (s, 18H); 13C NMR (100 MHz, CDCl3) δ 169.99, 160.40, 158.51, 158.39, 150.45, 150.32, 145.79, 145.53, 131.66, 130.85, 115.07, 114.64, 112.63, 110.45, 84.25, 51.01, 33.46, 28.48, 27.92, 26.77; LC method (C) Rt=4.122 min.
Di-tert-butyl (2-(2-((5-amino-2-methylphenyl)(propyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0105-2 (78%) as a yellow foamy solid. Rf=0.30 (DCM-MeOH, 95:5); 1H NMR (400 MHz, CDCl3) δ 8.72 (d, J=5.7 Hz, 1H), 7.47 (d, J=5.7 Hz, 1H), 7.37 (s, 1H), 7.11 (d, J=8.2 Hz, 1H), 6.68 (dd, J=8.1, 2.2 Hz, 1H), 6.59 (s, 1H), 3.96 (brs, 2H), 2.12 (s, 3H), 2.04 (s, 3H), 1.69-1.59 (m, 2H), 1.54 (s, 18H), 0.92 (t, J=7.3 Hz, 2H); LC method (D) Rt=5.050 min.
Di-tert-butyl (2-(2-((5-amino-2-methylphenyl)amino)thiazol-4-yl)-5-fluoropyrimidin-4-yl)carbamate OR0239-2 (70%) as a yellow powder. 1H NMR (400 MHz, CDCl3) δ 8.62 (s, 1H), 7.55 (s, 1H), 7.02 (d, J=8.0 Hz, 1H), 6.88 (d, J=2.0, 1H), 6.45 (dd, J=8.0, 2.0 Hz, 1H), 3.67 (brs, 2H), 2.19 (s, 3H), 1.45 (s, 18H); LC method (D) Rt=5.101 min.
Di-tert-butyl (2-(2-((5-nitro-2-methylphenyl)(propyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0105-3.
Under argon, at 0° C., to a stirred solution of di-tert-butyl (2-(2-((2-methyl-5-nitrophenyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate dCKi-1-3 (400 mg, 0.757 mmol) in a mixture of tetrahydrofuran-dimethylformamide (5:2, 14 mL) was added sodium hydride (60% in mineral oil, 38 mg, 0.95 mmol). After addition, the reaction mixture was stirred during 30 minutes at 0° C., allowed to warm to room temperature and stirred for further 30 minutes at this temperature. Then, propyl bromide (90 μL, 0.99 mmol) was added at 0° C. and the resulting suspension was heated to 60° C. for 3 days. The reaction was carefully quenched at 0° C. with methanol (10 ml) and concentrated under reduced pressure. The residue was purified by flash chromatography, gradient Cyclohexane-EtOAc (95:5 to 85:15) to afford di-tert-butyl (2-(2-((5-nitro-2-methylphenyl)(propyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0105-3 (142 mg, 33%) as a white solid. Rf=0.67 (PE-EtOAc, 1:1); 1H NMR (400 MHz, CDCl3) δ 8.72 (d, J=5.7 Hz, 1H), 8.18 (dd, J=8.4, 2.2 Hz, 1H), 8.16 (d, J=2.2 Hz, 1H), 7.53 (d, J=8.4 Hz, 1H), 7.52 (d, J=5.7 Hz, 1H), 7.43 (s, 1H), 4.09-3.96 (m, 2H), 2.37 (s, 3H), 1.72-1.59 (m, 2H), 1.54 (s, 18H), 0.95 (t, J=7.4 Hz, 3H); LCMS C2H34N6O6S method (B) Rt=8.987 min, ESI+ m/z=571.3 (M+H).
General Procedure for the Synthesis of di-tert-butyl (2-(2-((3-nitroaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate derivatives. To a stirred solution of appropriate di-tert-butyl (2-(2-bromoacetyl)-pyrimidin-4-yl)carbamate (1.3 mmol) and 1-(3-nitroaryl)thiourea (1.22 mmol) in ethanol (40 mL) was added potassium carbonate (258 mg, 1.87 mmol). The resulting mixture was stirred at temperature and for time indicated below. The solvent was distillated off under reduced pressure. The residue was purified by flash chromatography to afford the corresponding di-tert-butyl (2-(2-((3-nitroaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate.
Di-tert-butyl (2-(2-((2-methyl-5-nitrophenyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate dCKi-1-3 (96%) as a dark orange foamy solid. Rf=0.10 (PE-EtOAc, 1:1); 1H NMR (400 MHz, CDCl3) δ 8.58 (d, J=5.7 Hz, 1H), 8.47 (d, J=2.2 Hz, 1H), 7.90 (dd, J=8.3, 2.2 Hz, 1H), 7.60 (s, 1H), 7.55 (d, J=5.7 Hz, 1H), 7.38 (d, J=8.3 Hz, 1H), 2.40 (s, 3H), 1.56 (s, 18H); LCMS C24H28N6O6S, method (B) Rt=8.197 min, ESI+ m/z=529.2 (M+H).
Di-tert-butyl (2-(2-(8-nitro-2,3,4,5-tetrahydro-1H-benzo[b]azepin-1-yl)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0289-3 (50%) as a yellow foamy solid. Rf=0.20 (DCM-MeOH, 98:2); 1H NMR (400 MHz, CDCl3) δ 8.73 (d, J=5.8 Hz, 1H), 8.32 (d, J=2.2 Hz, 1H), 8.10 (dd, J=8.4, 2.2 Hz, 1H), 7.54 (d, J=5.8 Hz, 1H), 7.47 (d, J=8.5 Hz, 1H), 7.46 (s, 1H), 4.06 (brs, 2H), 2.91-2.82 (m, 2H), 2.01-1.88 (m, 2H), 1.75 (brs, 2H), 1.54 (s, 18H). 13C NMR (100 MHz, CDCl3) δ 168.86, 159.95, 158.57, 158.35, 150.25, 148.70, 147.50, 145.45, 131.81, 123.56, 123.51, 122.87, 113.21, 110.52, 84.37, 51.08, 34.24, 28.24, 27.92, 25.42; LCMS C27H32N6O6S, method (B) Rt=8.694 min, ESI+ m/z=569.3 (M+H).
Di-tert-butyl (5-fluoro-2-(2-((2-methyl-5-nitrophenyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0239-3 (83%) as a deep orange solid. 1H NMR (400 MHz, CDCl3) δ 8.67 (s, 1H), 8.52 (d, J=2.2 Hz, 1H), 7.92 (dd, J=8.3, 2.2 Hz, 1H), 7.71 (s, 1H), 7.40 (d, J=8.3 Hz, 1H), 2.42 (s, 3H), 1.46 (s, 18H); LCMS C24H27FN6O6S, method (A) Rt=7.871 min, ESI− m/z=545.2 (M−H).
General Procedure for the Synthesis of 1-(3-nitroaryl)thiourea Derivatives. A suspension of appropriate N-((3-nitroaryl)carbamothioyl)acetamide (2 mmol) and potassium carbonate (1.53 g, 11.1 mmol), in methanol (10 mL) was stirred at room temperature, upon complete consumption of the starting material monitored by TLC or LCMS. The solvent was distillated off under reduced pressure and the residue was poured onto water (30 mL), and then extracted with DCM (3×50 mL). The combined organic layers were dried over Na2SO4, and the solvent was distillated off under reduced pressure. The crude product was purified by flash chromatography to afford corresponding 1-(3-nitroaryl)thiourea.
1-(2-Methyl-5-nitrophenyl)thiourea dCKi-1-4 (90%) as a pale yellow solid. Rf=0.20 (PE-EtOAc, 1:1); 1H NMR (300 MHz, DMSO-d6) δ 9.41 (s, 1H), 8.27 (d, J=2.4 Hz, 1H), 7.98 (dd, J=8.4, 2.4 Hz, 1H), 7.70 (s, 2H), 7.51 (d, J=8.5 Hz, 1H), 2.31 (s, 3H); LCMS C8H9N3O2S method (D) Rt=4.675 min, ESI+ m/z=212.1 (M+H).
N-8-Nitro-2,3,4,5-tetrahydro-1H-benzo[b]azepine-1-carbothioamide OR0289-4 (79%) as a white solid. Rf=0.31 (DCM-MeOH, 98:2); 1H NMR (400 MHz, CDCl3) δ 8.20-8.12 (m, 2H), 7.51 (d, J=7.9 Hz, 1H), 5.62 (brs, 2H), 5.31 (d, J=12.8 Hz, 1H), 2.99 (t, J=12.8 Hz, 1H), 2.86 (m, 2H), 2.33 (m, 1H), 2.09 (d, J=14.0 Hz, 1H), 1.83 (d, J=13.9 Hz, 1H), 1.41 (m, 1H); LCMS C11H13N3O2S method (B) Rt=5.782 min, ESI+ m/z=252.1 (M+H).
General Procedure for the Synthesis of N-((3-nitroaryl)carbamothioyl)acetamide Derivatives. Under argon, to a suspension of potassium thiocyanate (982 mg, 10 mmol) in acetone (30 mL) was added dropwise acetyl chloride (0.60 mL, 7.95 mmol). The resulting solution was refluxed for 3 hrs. After to cooling to room temperature, a solution of appropriate 3-nitroaniline (7.68 mmol) in acetone (20 mL) was added dropwise. The mixture was stirred overnight at room temperature, upon complete consumption of the starting material monitored by TLC or LCMS. The solvent was distillated off under reduced pressure and the residue was poured onto water (100 mL) and stirred for 1 hr. The resulting precipitate was collected by filtration, to afford corresponding N-((3-nitroaryl)carbamothioyl)acetamide.
N-((2-Methyl-5-nitrophenyl)carbamothioyl)acetamide dCKi-1-5 (99%) as a dark orange powder. Rf=(PE-EtOAc, 85:15); 1H NMR (300 MHz, DMSO-d6) δ12.37 (s, 1H), 11.67 (s, 1H), 8.67 (d, J=2.4 Hz, 1H), 8.06 (dd, J=8.4, 2.4 Hz, 1H), 7.58 (d, J=8.5 Hz, 1H), 2.33 (s, 3H), 2.18 (s, 3H); LCMS C10H11N3O3S method (B) Rt=6.036 min, ESI+ m/z=254.1 (M+H).
N-(8-Nitro-2,3,4,5-tetrahydro-1H-benzo[b]azepine-1-carbonothioyl)acetamide OR0289-5 (93%) as a light brown solid. LCMS C13H15N3O3S method (B) Rt=6.061 min, ESI+ m/z=294.1 (M+H).
8-Nitro-2,3,4,5-tetrahydro-1H-benzo[b]azepine OR0289-6
At −5° C., under stirring, to concentrated sulfuric acid (30 mL) was added by solid fraction over 30 min, a mixture of 2,3,4,5-tetrahydro-1H-benzo[b]azepine (2.94 g, 20 mmol) and potassium nitrate (2.02 g, 20 mmol). After addition, the reaction mixture was poured onto ice (100 g), then Na2CO3 was added until pH=8-9, and extracted with DCM (3×20 mL). The combined organic layers were dried over Na2SO4 and the solvent was distillated off under reduced pressure. The residue was purified by flash chromatography, DCM-PE (8:2) to afford 8-nitro-2,3,4,5-tetrahydro-1H-benzo[b]azepine OR0289-6 (2.70 g, 71%) as an orange oil which crystallized from a mixture of MeOH-Et2O-PE as orange plates. Rf=0.35 (DCM-PE, 8:2); 1H NMR (400 MHz, CDCl3) δ 7.67 (dd, J=8.2, 2.3 Hz, 1H), 7.64 (d, J=2.3 Hz, 1H), 7.22 (d, J=8.2 Hz, 1H), 3.15-3.08 (m, 2H), 2.91-2.82 (m, 2H), 1.89-1.83 (m, 2H), 1.74-1.62 (m, 2H); LCMS C10H12N2O2 method (B) Rt=6.522 min, ESI+ m/z=193.1 (M+H).
General Procedure for the Synthesis of di-tert-butyl (2-(2-bromoacetyl)-pyrimidin-4-yl)carbamate Derivatives. To a stirred solution of appropriate di-tert-butyl (2-(1-ethoxyvinyl)pyrimidin-4-yl)carbamate (4 mmol) in a mixture of tetrahydrofuran-water (3:1, 8 mL) was added in one portion N-bromosuccinimide (844 mg, 4.74 mmol). The resulting solution was stirred for 3 hrs at room temperature, upon complete consumption of the starting material. The reaction mixture was concentrated under reduced pressure without heating (T≤30° C.). The crude product was crystallized from Et2O to afford corresponding di-tert-butyl (2-(2-bromoacetyl)-pyrimidin-4-yl)carbamate.
Di-tert-butyl (2-(2-bromoacetyl)-pyrimidin-4-yl)carbamate dCKi-1-6 (95%) as a white crystals. Rf=0.35 (PE-EtOAc, 8:2); 1H NMR (400 MHz, CDCl3) δ 8.78 (d, J=5.8 Hz, 1H), 7.93 (d, J=5.8 Hz, 1H), 4.68 (s, 2H), 1.57 (s, 18H); LCMS C16H22BrN3O5 method (B) Rt=7.009 min, ESI+ m/z=418.2 (M+H).
Di-tert-butyl (2-(2-bromoacetyl)-5-fluoropyrimidin-4-yl)carbamate OR0239-4 (quantitative) as a white crystals engaged in the next step without further purification. LCMS C16H21BrFN3O5 method (A) Rt=7.256 min, ESI+ m/z=234.1 (M-(Boc)2).
General Procedure for the Synthesis of di-tert-butyl (2-(1-ethoxyvinyl)pyrimidin-4-yl)carbamate Derivatives. Under argon, to a solution of appropriate di-tert-butyl (2-chloropyrimidin-4-yl)carbamate (5.0 mmol) in dry 1,4-dioxane or toluene (50 mL) were added successively tributyl(1-ethoxyvinyl)tin (2.52 g, 7.0 mmol) and cesium fluoride (1.51 g, 10.0 mmol). The resulting mixture was thoroughly degassed several times under argon fillings before addition of bis(triphenylphosphine)palladium dichloride (351 mg, 0.5 mmol, 10 mol %). The resulting mixture was warmed until complete consumption of starting material. After cooling to room temperature the resulting mixture was filtrated through a short pad of Celite, rinsing with EtOAc (50 mL). The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography to afford corresponding di-tert-butyl (2-(1-ethoxyvinyl)pyrimidin-4-yl)carbamate.
Di-tert-butyl (2-(1-ethoxyvinyl)pyrimidin-4-yl)carbamate dCKi-1-7. Under argon, to a solution of di-tert-butyl (2-chloropyrimidin-4-yl)carbamate dCKi-1-8 (25.5 g, 77.4 mmol) in dry 1,4-dioxane (450 mL) were added successively tributyl(1-ethoxyvinyl)tin (38.4 g, 106.2 mmol) and cesium fluoride (23.3 g, 153.0 mmol). The resulting mixture was thoroughly degassed several times under argon fillings before addition of bis(triphenylphosphine)palladium dichloride (5.5 g, 7.75 mmol, 10 mol %). The resulting mixture was stirred at 90° C. overnight, and then allowed to cool to room temperature before filtration through a pad of Celite, washing with EtOAc (800 mL). The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography, PE-EtOAc (8:2) to afford di-tert-butyl (2-(1-ethoxyvinyl)pyrimidin-4-yl)carbamate dCKi-1-7 (20.7 g, 73%) as yellow crystals. Rf=0.35 (PE-EtOAc, 8:2); 1H NMR (400 MHz, CDCl3) δ 8.66 (d, J=5.7 Hz, 1H), 7.56 (d, J=5.7 Hz, 1H), 5.58 (d, J=1.9 Hz, 1H), 4.57 (d, J=1.9 Hz, 1H), 3.99 (q, J=7.0 Hz, 2H), 1.55 (s, 18H), 1.46 (t, J=7.0 Hz, 3H); LCMS C18H27N3O5 method (A) Rc=7.608 min, ESI+ m/z=366.3 (M+H).
Di-tert-butyl (2-(1-ethoxyvinyl)-5-fluoropyrimidin-4-yl)carbamate OR0239-5. A stirring solution of di-tert-butyl (2-chloro-5-fluoropyrimidin-4-yl)carbamate OR0239-6 (400 mg, 1.15 mmol), tributyl(1-ethoxyvinyl)tin (550 μL, 1.61 mmol), copper(I) bromide (25 mg, 0.17 mmol) and cesium fluoride (350 mg, 2.3 mmol) in dry toluene (10 mL) was degassed under argon fillings for 15 min. PdCl2(PPh3)2 (41 mg, 0.06 mmol) and triphenylphosphine (45 mg, 0.17 mmol) were successively added. Argon was bubbled through for another 5 min and the mixture was refluxed during 4 hrs upon complete conversion monitored by TLC. The resulting mixture was allowed to cool to room temperature, filtered through a pad of celite and rinsed with DCM. The crude product was purified by flash chromatography, DCM-EtOAc to afford di-tert-butyl (2-(1-ethoxyvinyl)-5-fluoropyrimidin-4-yl)carbamate OR0239-5 as a colorless oil (401 mg, 91%). 1H NMR (400 MHz, CDCl3) δ 8.62 (d, J=1.4 Hz, 1H), 5.60 (d, J=2.2 Hz, 1H), 4.62 (d, J=2.2 Hz, 1H), 4.02 (q, J=7.0 Hz, 2H), 1.48 (t, J=7.0 Hz, 3H), 1.44 (s, 18H); LCMS C18H26FN3O5 method (A) Rt=7.568 min, ESI+ m/z=384.2 (M+H).
General Procedure for the Synthesis of di-tert-butyl (2-chloropyrimidin-4-yl)carbamate Derivatives. To a stirred solution of appropriate 2-chloro-4-aminopyrimidine (1.95 g, 15 mmol) in tetrahydrofuran (50 mL), was added di-tert-butyl dicarbonate (8.0 g, 37 mmol) and 4-dimethylaminopyridine (124 mg, 1 mmol). The resulting solution was stirred at room temperature overnight upon completion of the reaction. The solvent was distillated off under reduced pressure and the residue was diluted with DCM (50 mL) and washed with H2O (2×10 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure to afford corresponding di-tert-butyl (2-chloropyrimidin-4-yl)carbamate.
Di-tert-butyl (2-chloropyrimidin-4-yl)carbamate dCKi-1-8 (quantitative) as a white solid. Rf=0.70 (PE-EtOAc, 85:15). 1H NMR (300 MHz, CDCl3) δ 8.46 (d, J=5.7 Hz, 1H), 7.71 (d, J=5.7 Hz, 1H), 1.56 (s, 18H); LCMS C14H20ClN3O4 method (B) Rt=7.407 min, ESI+ m/z=330.2 (M+H).
Di-tert-butyl (2-chloro-5-fluoropyrimidin-4-yl)carbamate OR0239-6 (28%) as a white powder. 1H NMR (400 MHz, CDCl3) δ 8.50 (s, 1H), 1.46 (s, 18H); LCMS C14H19ClFN3O4 method (A) Rt=7.759 min, ESI+ m/z=192.1 (M-(Boc)2+HCO2H).
N-([1,1′-biaryl]-3-yl)-4-(4-aminopyrimidin-2-yl)thiazol-2-amine derivatives were prepared by convergent synthesis, in eight steps, from commercially available 4-amino-2-chloropyrimidine and appropriate bromo-3-nitrobenzene (Scheme 2). The 4-amino-2-chloropyrimidine was protected using Boc2O to afford corresponding bis-carbamate. A Stille cross-coupling reaction with tributyl(1-ethoxyvinyl)tin gave the enol ether which was then turned into the corresponding α-bromoketone using N-bromosuccinimide. Starting from appropriate bromo-3-nitrobenzene, a Suzuki cross-coupling reaction with corresponding aryl boronate allowed introducing the key biaryl scaffold. Hydrogenation of the nitro group followed by condensation with potassium thiocyanate, after optional (C1-C6)alkylation, gave the corresponding thiourea. As previously described, the α-bromoketone was engaged in the Hantzsch thiazole synthesis with the thiourea leading to the corresponding thiazole. Finally, deprotection with TFA led to expected N-([1,1′-biaryl]-3-yl)-4-(4-aminopyrimidin-2-yl)thiazol-2-amines.
General Procedure for the Synthesis of N-([1,1′-biaryl]-3-yl)-4-(4-aminopyrimidin-2-yl)thiazol-2-amine. As previously described for N-(3-((4-(4-aminopyrimidin-2-yl)thiazol-2-yl)amino)-4-aryl)benzamide derivatives.
4-(4-Aminopyrimidin-2-yl)-N-(4-methyl-4′-((4-methylpiperazin-1-yl)methyl)-[1,1′-biphenyl]-3-yl)thiazol-2-amine OR0274 (57%) as a white powder. 1H NMR (400 MHz, CDCl3) δ 8.31 (d, J=5.7 Hz, 1H), 7.76 (s, 1H), 7.59 (s, 1H), 7.55 (d, J=8.1 Hz, 2H), 7.39 (d, J=8.1 Hz, 2H), 7.32-7.27 (m, 2H), 6.34 (d, J=5.7 Hz, 1H), 4.98 (brs, 2H), 3.55 (s, 2H), 2.50 (brs, 8H), 2.34 (s, 3H), 2.30 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 166.65, 163.11, 160.54, 156.49, 150.57, 140.32, 139.31, 139.17, 137.56, 131.73, 129.81, 128.70, 126.88, 123.25, 118.96, 110.94, 103.41, 62.75, 55.20, 53.11, 46.07, 17.69. LC method (D) Rt=3.558 min.
4-(4-Aminopyrimidin-2-yl)-N-(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)thiazol-2-amine OR0325 (45%) as a white powder. 1H NMR (400 MHz, CDCl3) δ 8.30 (d, J=5.7 Hz, 1H), 7.74 (s, 1H), 7.58 (s, 1H), 7.52 (d, J=8.2 Hz, 2H), 7.35 (brs, 1H), 7.29-7.27 (m, 4H), 6.33 (d, J=5.7 Hz, 1H), 4.99 (s, 2H), 2.85 (dd, J=9.9, 6.4 Hz, 1H), 2.65 (dd, J=9.9, 6.4 Hz, 1H), 2.52 (brs, 8H), 2.33 (s, 3H), 2.32 (s, 3H). 13C NMR (100 MHz, CDCl3) δ 166.66, 163.10, 160.53, 156.48, 150.57, 140.34, 139.73, 139.16, 138.35, 131.71, 129.32, 128.62, 127.05, 123.19, 118.92, 110.92, 103.40, 60.47, 55.19, 53.20, 46.11, 33.31, 17.67; LC method (D) Rt=3.602 min. 3′-((4-(4-Aminopyrimidin-2-yl)thiazol-2-yl)amino)-4′-methyl-[1,1′-biphenyl]-4-ol OR0320 (58%) as a white powder. 1H NMR (400 MHz, MeOD) δ 8.12 (d, J=6.0 Hz, 1H), 7.76 (d, J=1.6 Hz, 1H), 7.49 (s, 1H), 7.46 (d, J=8.6 Hz, 2H), 7.32 (dd, J=8.1, 1.6 Hz, 1H), 7.29 (d, J=8.1 Hz, 1H). 6.84 (d, J=8.6 Hz, 2H), 6.43 (d, J=6.0 Hz, 1H), 2.34 (s, 3H). 13C NMR (100 MHz, MeOD) δ 169.65, 165.60, 160.77, 158.24, 154.87, 150.72, 141.44, 140.88, 133.12, 132.59, 130.83, 128.88, 124.35, 122.07, 116.64, 112.10, 104.58, 17.61; LC method (D) Rt=4.369 min.
3′-((4-(4-Aminopyrimidin-2-yl)thiazol-2-yl)amino)-4′-methyl-[1,1′-biphenyl]-4-ol OR0321 (53%) as a white powder. 1H NMR (400 MHz, CDCl3) δ 8.28 (s, 1H), 7.70 (s, 1H), 7.57 (s, 1H), 7.28-7.26 (m, 2H), 7.12-6.96 (m, 3H), 6.33 (d, J=5.5 Hz, 1H), 5.03 (brs, 2H), 3.94 (s, 3H), 2.32 (s, 3H); LC method (D) Rt=4.389 min.
4-(4-Aminopyrimidin-2-yl)-N-(4-methyl-4′-(3-(4-methylpiperazin-1-yl)propyl)-[1,1′-biphenyl]-3-yl)thiazol-2-amine OR0331 (46%) as a light yellow powder. 1H NMR (400 MHz, CDCl3) δ 8.26 (d, J=5.7 Hz, 1H), 7.74 (brs, 1H), 7.56 (s, 1H), 7.51-7.49 (m, 3H), 7.31-7.24 (m, 4H), 6.31 (d, J=5.7 Hz, 1H), 5.05 (brs, 2H), 2.66 (t, J=7.8 Hz, 2H), 2.47 (brs, 8H), 2.39 (t, J=7.8 Hz, 2H), 2.31 (s, 3H), 2.28 (s, 3H), 1.81 (quint, J=7.8 Hz, 2H); 13C NMR (100 MHz, CDCl3) δ 166.67, 163.10, 160.55, 156.51, 150.56, 141.65, 140.45, 139.12, 138.04, 131.71, 129.02, 128.51, 126.97, 123.20, 118.89, 110.97, 103.40, 58.08, 55.26, 53.29, 46.16, 33.47, 28.71, 17.68; LCMS C28H33N7S method (B) Rt=3.881 min, ESI+ m/z=500.1 (M+H).
4-(4-Aminopyrimidin-2-yl)-N-(4-methyl-4′-(3-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)-N-propylthiazol-2-amine OR0345 (42%), as a colorless solid; 1H NMR (400 MHz, CDCl3) δ 8.26 (d, J=5.7 Hz, 1H), 7.53-7.46 (m, 4H), 7.39-7.38 (m, 2H), 7.28-7.25 (m, 2H), 6.25 (d, J=5.7 Hz, 1H), 5.3 (brs, 2H), 4.01 (brs, 2H), 2.86-2.82 (m, 2H), 2.87-2.51 (m, 2H), 2.52 (brs, 8H), 2.31 (s, 3H), 2.26 (s, 3H), 1.66 (sext, J=7.3 Hz, 2H), 0.92 (t, J=7.3 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 171.27, 163.14, 160.88, 156.41, 151.19, 143.27, 140.74, 139.81, 137.84, 136.17, 132.49, 129.33, 127.97, 126.96, 111.21, 103.18, 60.39, 55.13, 53.61, 53.11, 46.05, 33.25, 21.27, 17.45, 11.40; LCMS C30H37N7S method (B) Rt=4.152 min, ESI+m/z=528.3 (M+H).
4-(4-Aminopyrimidin-2-yl)-N-(4-methyl-4′-(3-(4-methylpiperazin-1-yl)propyl)-[1,1′-biphenyl]-3-yl)-N-propylthiazol-2-amine OR0598 (70%), as a pale yellow solid. Rf=0.40 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 8.29 (d, J=5.8 Hz, 1H), 7.53 (dd, J=7.9, 1.7 Hz, 1H), 7.49 (d, J=8.1 Hz, 2H), 7.46 (d, J=1.7 Hz, 1H), 7.39 (d, J=7.6 Hz, 1H), 7.37 (s, 1H), 7.24 (d, J=8.1 Hz, 2H), 6.29 (d, J=5.8 Hz, 1H), 5.10 (s, 2H), 4.02 (brs, 2H), 2.72-2.65 (m, 2H), 3.11-2.55 (m, 8H), 2.54-2.45 (m, 2H), 2.43 (s, 3H), 2.27 (s, 3H), 1.89 (quint, J=7.6 Hz, 2H), 1.67 (sext, J=7.5 Hz, 2H), 0.93 (t, J=7.5 Hz, 3H). 13C NMR (100 MHz, CDCl3) δ 171.41, 163.09, 160.94, 156.53, 151.17, 143.29, 141.14, 140.78, 137.78, 136.24, 132.54, 129.04, 128.02, 127.00, 111.31, 103.19, 57.48, 54.33, 53.53, 52.03, 45.29, 33.18, 28.07, 21.32, 17.47, 11.44; LCMS C31H39N7S method (B) Rt=4.253 min, ESI+ m/z=542.3 (M+H).
2-(2-(8-(4-(2-(4-Methylpiperazin-1-yl)ethyl)phenyl)-2,3,4,5-tetrahydro-1H-benzo[b]azepin-1-yl)thiazol-4-yl)pyrimidin-4-amine OR0402 (87%), as a pale yellow solid. Rf=0.18 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 8.12 (d, J=6.0 Hz, 1H), 7.60 (d, J=1.8 Hz, 1H), 7.56-7.47 (m, 3H), 7.38 (d, J=7.9 Hz, 1H), 7.35 (s, 1H), 7.27 (d, J=8.2 Hz, 2H), 6.43 (d, J=6.0 Hz, 1H), 4.03 (brs, 2H), 2.85-2.79 (m, 2H), 2.78-2.72 (m, 2H), 2.66-2.58 (m, 2H), 2.55 (brs, 8H), 2.30 (s, 3H), 1.95-1.93 (m, 2H), 1.75-7.73 (m, 2H). 13C NMR (100 MHz, MeOD) δ 170.97, 165.49, 161.54, 155.76, 151.89, 146.59, 142.09, 141.04, 140.70, 139.12, 132.64, 130.35, 127.88, 127.59, 127.25, 111.86, 104.49, 61.16, 55.53, 53.55, 52.04, 45.89, 35.03, 33.63, 29.59, 27.58; LCMS C30H35N7S method (B) Rt=4.064 min, ESI+m/z=526.3 (M+H).
4-(4-Aminopyrimidin-2-yl)-N-(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethoxy)-[1,1′-biphenyl]-3-yl)-N-propylthiazol-2-amine OR0596 (63%), as a white solid. 1H NMR (400 MHz, CDCl3) δ 8.29 (d, J=5.8 Hz, 1H), 7.50 (d, J=8.8 Hz, 2H), 7.49 (dd, J=7.7, 1.8 Hz, 1H), 7.43 (d, J=2.0 Hz, 1H), 7.38 (s, 1H), 7.37 (d, J=7.7 Hz, 1H), 6.97 (d, J=8.8 Hz, 2H), 6.27 (d, J=5.8 Hz, 1H), 5.12 (s, 2H), 4.14 (t, J=5.8 Hz, 2H), 4.02 (brs, 2H), 2.83 (t, J=5.8 Hz, 2H), 2.63 (brs, 4H), 2.48 (brs, 4H), 2.29 (s, 3H), 2.26 (s, 3H), 1.67 (sext, J=7.4 Hz, 2H), 0.93 (t, J=7.4 Hz, 3H). 13C NMR (100 MHz, CDCl3) δ 171.39, 163.09, 160.98, 158.62, 156.58, 151.19, 143.28, 140.59, 135.71, 132.73, 132.49, 128.02, 127.70, 126.75, 115.12, 111.27, 103.16, 66.17, 57.28, 55.16, 53.73, 53.56, 46.16, 21.31, 17.44, 11.44; LCMS C30H37N7S method (B) Rt=4.304 min, ESI+ m/z=544.2 (M+H).
4-(4-Aminopyrimidin-2-yl)-N-(4-methyl-4′-(2-(4-methylpiperazin-1-yl)-2-oxoethoxy)-[1,1′-biphenyl]-3-yl)-N-propylthiazol-2-amine OR0597 (49%), as a white solid. 1H NMR (400 MHz, CDCl3) δ 8.30 (d, J=5.8 Hz, 1H), 7.51 (d, J=8.8 Hz, 2H), 7.48 (dd, J=8.0, 1.9 Hz, 1H), 7.42 (d, J=1.9 Hz, 1H), 7.38 (s, 1H), 7.38 (d, J=8.0 Hz, 1H), 7.01 (d, J=8.8 Hz, 2H), 6.29 (d, J=5.8 Hz, 1H), 5.04 (s, 2H), 4.72 (s, 2H), 4.03 (brs, 2H), 3.63 (m, 4H), 2.40 (m, 4H), 2.29 (s, 3H), 2.27 (s, 3H), 1.67 (sext, J=7.4 Hz, 2H), 0.94 (t, J=7.4 Hz, 3H). 13C NMR (100 MHz, CDCl3) δ 171.41, 166.40, 163.07, 161.00, 157.69, 156.63, 151.19, 143.30, 140.40, 135.97, 133.65, 132.55, 128.22, 127.81, 126.83, 115.16, 111.31, 103.16, 67.85, 55.27, 54.74, 53.51, 46.15, 45.39, 42.18, 21.33, 17.45, 11.45; LCMS C30H35N7O2S method (B) Rt=4.296 min, ESI+ m/z=558.2 (M+H).
4-(4-Aminopyrimidin-2-yl)-N-(2-methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethyl)pyridin-3-yl)phenyl)-N-propylthiazol-2-amine OR0599 (10%) as a yellow solid. Rf=0.40 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 8.74 (s, 1H), 8.30 (d, J=5.3 Hz, 1H), 7.77 (d, J=7.9 Hz, 1H), 7.52 (d, J=7.9 Hz, 1H), 7.45 (s, 1H), 7.43 (d, J=7.9 Hz, 1H), 7.39 (s, 1H), 7.24 (d, J=7.9 Hz, 1H), 6.29 (d, J=5.3 Hz, 1H), 5.07 (s, 2H), 4.04 (brs, 2H), 3.03 (m, 2H), 2.80 (m, 2H), 2.60 (brs, 4H), 2.48 (brs, 4H), 2.29 (s, 6H), 1.66 (sext, J=7.2 Hz, 2H), 0.94 (t, J=7.2 Hz, 3H). 13C NMR (100 MHz, CDCl3) δ 171.20, 163.08, 160.97, 159.62, 156.65, 151.28, 147.57, 143.60, 137.68, 137.31, 134.70, 133.15, 132.92, 128.14, 127.01, 123.28, 111.35, 103.20, 58.43, 55.24, 53.54, 53.16, 46.15, 35.53, 21.36, 17.56, 11.44; LCMS C29H36N8S method (B) Rt=3.950 min, ESI+ m/z=529.4 (M+H).
General Procedure for the Synthesis of di-tert-butyl (2-(2-([1,1′-biaryl]-3-ylamino)thiazol-4-yl)pyrimidin-4-yl)carbamate Derivatives. As previously described for di-tert-butyl (2-(2-((3-nitroaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate derivatives.
Di-tert-butyl (2-(2-((4-methyl-4′-((4-methylpiperazin-1-yl)methyl)-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0274-1 (37%) as a light brown powder. 1H NMR (400 MHz, CDCl3) δ 8.70 (d, J=5.7 Hz, 1H), 7.75 (s, 1H), 7.57-7.50 (m, 4H), 7.39 (d, J=8.1 Hz, 2H), 7.31 (s, 2H), 3.55 (s, 2H), 2.50 (brs, 8H), 2.34 (s, 3H), 2.30 (s, 3H), 1.56 (s, 18H).
Di-tert-butyl (2-(2-((4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0325-1 (46%) as a light brown powder. 1H NMR (400 MHz, CDCl3) δ 8.70 (d, J=5.7 Hz, 1H), 7.73 (s, 1H), 7.53-7.51 (m, 4H), 7.33-7.26 (m, 4H), 2.85 (dd, J=9.9, 6.4 Hz, 2H), 2.65 (dd, J=9.9, 6.4 Hz, 1H), 2.53 (brs, 8H), 2.34 (s, 3H), 2.32 (s, 3H), 1.56 (s, 18H); LC method (C) Rt=3.933 min.
Di-tert-butyl (2-(2-((4′-hydroxy-4-methyl-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0320-1 (48%) as a light brown powder. 1H NMR (400 MHz, CDCl3) δ 8.68 (d, J=5.7 Hz, 1H), 7.66 (s, 1H), 7.53 (d, J=5.7 Hz, 1H), 7.50 (s, 1H), 7.46 (d, J=8.6 Hz, 2H), 7.27-7.25 (m, 1H), 6.92 (d, J=8.6 Hz, 2H), 2.30 (s, 3H), 1.56 (s, 18H); LC method (A) Rt=8.406 min.
Di-tert-butyl (2-(2-((4′-hydroxy-3′-methoxy-4-methyl-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0321-1 (52%) as a light brown powder. 1H NMR (400 MHz, CDCl3) δ 8.70 (d, J=5.6 Hz, 1H), 7.70 (s, 1H), 7.54 (d, J=5.6 Hz, 1H), 7.51 (s, 1H), 7.28-7.26 (m 2H), 7.13-7.05 (m, 2H), 6.99 (d, J=8.1 Hz, 1H), 5.65 (brs, 1H), 3.97 (s, 3H), 2.34 (s, 3H), 1.56 (s, 18H); LCMS C31H35N5O6S method (A) Rt=8.452 min, ESI+ m/z=606.3 (M+H).
Di-tert-butyl (2-(2-((4-methyl-4′-(2-(4-methylpiperazin-1-yl)propyl)-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0331-1 (58%) as a yellow powder. LCMS C38H49N7O4S method (B) Rt=5.470 min, ESI+ m/z=700.4 (M+H).
Di-tert-butyl (2-(2-((4-methyl-4′-(3-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0345-1 (47%) as a yellow powder. Rf=0.42 (DCM-MeOH—NH4OH, 90:9:1). 1H NMR (400 MHz, CDCl3) δ 8.73 (d, J=5.7 Hz, 1H), 7.55-7.46 (m, 4H), 7.41-7.39 (m, 2H), 7.31-7.24 (m, 3H), 5.3 (brs, 2H), 3.99 (brs, 2H), 2.86-2.50 (m, 12H), 2.28 (s, 3H), 2.17 (s, 3H), 1.69 (sext, J=7.4 Hz, 2H), 1.56 (s, 18H), 0.94 (t, J=7.4 Hz, 3H); LCMS C40H53N7O4S method (B) Rt=5.040 min, ESI+ m/z=728.4 (M+H).
Di-tert-butyl (2-(2-((4-methyl-4′-(3-(4-methylpiperazin-1-yl)propyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0598-1 (85%) as a light brown powder. Rf=0.42 (DCM-MeOH—NH4OH., 90:9:1). 1H NMR (400 MHz, CDCl3) δ 8.73 (d, J=5.7 Hz, 1H), 7.54 (dd, J=7.9, 1.7 Hz, 1H), 7.50 (d, J=8.6 Hz, 2H), 7.46-7.48 (m, 2H), 7.40 (d, J=8.0 Hz, 1H), 7.37 (s, 1H), 7.24 (d, J=8.2 Hz, 2H), 4.03 (brs, 2H), 2.94 (s, 8H), 2.96-2.87 (m, 2H), 2.70 (t, J=7.4 Hz, 2H), 2.57 (s, 3H), 2.28 (s, 3H), 1.98-1.94 (m, 2H), 1.68 (sext, J=7.4 Hz, 2H), 1.54 (s, 18H), 0.94 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 171.50, 160.50, 158.62, 158.55, 150.61, 150.37, 143.29, 140.75, 137.98, 136.32, 132.62, 129.04, 128.10, 127.23, 127.10, 112.49, 110.62, 84.24, 57.02, 55.86, 53.56, 53.36, 51.06, 32.95, 28.00, 27.96, 21.32, 17.46, 11.48; LCMS C41H55N7O4S method (B) Rt=5.956 min, ESI+ m/z=742.4 (M+H).
Di-tert-butyl (2-(2-(8-(4-(2-(4-methylpiperazin-1-yl)ethyl)phenyl)-2,3,4,5-tetrahydro-1H-benzo[b]azepin-1-yl)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0402-1 (60%) as a light brown powder. Rf=0.42 (DCM-MeOH—NH4OH, 90:9:1); LCMS C40H51N7O4S method (B) Rt=5.603 min, ESI+ m/z=726.3 (M+H).
Di-tert-butyl (2-(2-((4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethoxy)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0596-1 (94%) as a colorless oil. Rf=0.42 (DCM-MeOH—NH4OH, 90:9:1); LCMS C40H53N7O5S method (B) Rt=5.949 min, ESI+ m/z=744.4 (M+H).
Di-tert-butyl (2-(2-((4-methyl-4′-(2-(4-methylpiperazin-1-yl)-2-oxoethoxy)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0597-1 (82%) as a light brown solid. Rf=0.42 (DCM-MeOH—NH4OH, 90:9:1); LCMS C40H53N7O5S method (B) Rt=5.889 min, ESI+ m/z=758.2 (M+H).
Di-tert-butyl (2-(2-((2-methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethyl)pyridin-3-yl)phenyl)(propyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0599-1 (67%) as a light brown powder. Rf=0.42 (DCM-MeOH—NH4OH, 90:9:1); LCMS C39H52NSO4S method (B) Rt=5.489 min, ESI+ m/z=729.6 (M+H).
General Procedure for the Synthesis of 1-([1,1′-biaryl]-3-yl)thiourea Derivatives. Method (A): as previously described for 1-(3-nitroaryl)thiourea derivatives. Method (B): to a solution of appropriate [1,1′-biaryl]-3-amine (0.70 mmol) in 1N aqueous hydrochloric acid solution (7 mL) was added potassium thiocyanate (312 mg, 3.2 mmol). The resulting solution was successively stirred at 90° C. for 48 hours, allowed to cool to room temperature, then saturated aqueous NaHCO3 solution was added until pH=8, and extracted with EtOAc (3×50 mL). The combined organic layers were dried over Na2SO4, and the solvent was distillated off under reduced pressure. The residue was purified by chromatography to afford corresponding 1-([1,1′-biaryl]-3-yl)thiourea.
1-(4-Methyl-4′-((4-methylpiperazin-1-yl)methyl)-[1,1′-biphenyl]-3-yl)thiourea OR0274-2 (method A, 39%) as a white powder. 1H NMR (400 MHz, MeOD) δ 7.59 (d, J=8.3 Hz, 2H), 7.52 (dd, J=7.9, 1.9 Hz, 1H), 7.49 (d, J=1.9 Hz, 1H), 7.40 (d, J=8.3 Hz, 2H), 7.37 (d, J=7.9 Hz, 1H), 4.58 (s, 2H), 2.54 (brs, 8H), 2.31 (s, 3H), 2.30 (s, 3H).
1-(4-Methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)thiourea OR0325-2 (method A, 77%) as a white powder. 1H NMR (400 MHz, MeOD) δ 7.59-7.46 (m, 4H), 7.40-7.35 (m, 1H), 7.30-7.27 (m, 2H), 2.88-2.82 (m, 2H), 2.66-2.62 (m, 2H), 2.56 (brs, 8H), 2.31 (s, 6H); LC method (D) Rt=3.256 min.
1-(4′-Hydroxy-4-methyl-[1,1′-biphenyl]-3-yl)thiourea OR0320-2 (method A, 86%) as a white powder. 1H NMR (400 MHz, MeOD) δ 7.46-7.42 (m, 3H), 7.40 (d, J=1.4 Hz, 1H), 7.32 (d, J=8.0 Hz, 1H), 6.85 (d, J=8.7 Hz, 2H), 2.29 (s, 3H); LCMS C14H14N2OS method (A) Rt=5.366 min, ESI− m/z=257.4 (M−H).
1-(4′-Hydroxy-3′-methoxy-4-methyl-[1,1′-biphenyl]-3-yl)thiourea OR0321-2 (method A, 74%) as a white powder. 1H NMR (400 MHz, MeOD) δ 7.47 (dd, J=7.9, 1.9 Hz, 1H), 7.42 (d, J=1.9 Hz, 1H), 7.33 (d, J=7.9 Hz, 1H), 7.16 (d, J=2.1 Hz, 1H), 7.06 (dd, J=8.2, 2.1 Hz, 1H), 6.85 (d, J=8.2 Hz, 1H), 3.92 (s, 3H), 2.30 (s, 3H); LCMS C15H16N2O2S method (A) Rt=5.448 min, ESI+ m/z=289.1 (M+H).
1-(4-Methyl-4′-(2-(4-methylpiperazin-1-yl)propyl)-[1,1′-biphenyl]-3-yl)thiourea OR0331-2 (method B, 73%) as a pale yellow solid. LCMS C22H30N4S method (B) Rt=7.139 min, ESI+m/z=383.3 (M+H).
1-(4-Methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)-1-propylthiourea OR0345-2 (method B, quantitative) as a yellow powder. 1H NMR (400 MHz, CDCl3) δ 7.53 (dd, J=7.9, 1.9 Hz, 1H), 7.46 (d, J=8.2 Hz, 2H), 7.39 (d, J=7.9 Hz, 1H), 7.34 (d, J=1.9 Hz, 1H), 7.29 (d, J=8.2 Hz, 2H), 5.47 (brs, 2H), 4.46-4.42 (m, 1H), 3.86-3.79 (m, 1H), 2.87-2.82 (m, 2H), 2.65-2.59 (m, 2H), 2.51 (brs, 8H), 2.33 (s, 3H), 2.31 (s, 3H), 1.87-1.78 (m, 2H), 0.92 (t, J=7.4 Hz, 3H); LCMS C24H34N4S method (C) Rt=3.455 min, ESI+ m/z=411.3 (M+H).
1-(4-Methyl-4′-(3-(4-methylpiperazin-1-yl)propyl)-[1,1′-biphenyl]-3-yl)-1-propylthiourea OR0598-2 (method A, 90%) as a pale yellow solid. Rf=0.36 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.53 (dd, J=7.9, 1.8 Hz, 1H), 7.46 (d, J=8.1 Hz, 2H), 7.38 (d, J=7.9 Hz, 1H), 7.34 (d, J=1.8 Hz, 1H), 7.26 (d, J=8.1 Hz, 2H), 5.51 (brs, 2H), 4.51-4.41 (m, 1H), 3.72-3.61 (m, 1H), 2.72-2.64 (m, 2H), 2.56 (brs, 8H), 2.46-2.40 (m, 2H), 2.33 (s, 3H), 2.26 (s, 3H), 1.87 (quint, J=7.7 Hz, 2H), 1.86-1.62 (m, 2H), 0.92 (t, J=7.4 Hz, 3H); LCMS C25H36N4S method (B) Rt=4.417 min, ESI+ m/z=425.3 (M+H). 8-(4-(2-(4-Methylpiperazin-1-yl)ethyl)phenyl)-2,3,4,5-tetrahydro-1H-benzo[b]azepine-1-carbothioamide OR0402-2 (method A, 92%) used in the next step without neither work-up nor purification. LCMS C24H32N4S method (B) Rt=3.405 min, ESI+ m/z=409.3 (M+H).
1-(4-Methyl-4′-(2-(4-methylpiperazin-1-yl)ethoxy)-[1,1′-biphenyl]-3-yl)-1-propylthiourea OR0596-2 (method A, 61%) as a white solid. Rf=0.36 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.50 (dd, J=8.0, 1.8 Hz, 1H), 7.46 (d, J=8.8 Hz, 2H), 7.36 (d, J=8.0 Hz, 1H), 7.31 (d, J=1.8 Hz, 1H), 6.98 (d, J=8.8 Hz, 2H), 5.49 (brs, 2H), 4.49-4.41 (m, 1H), 4.15 (t, J=5.8 Hz, 2H), 3.68-3.61 (m, 1H), 2.84 (t, J=5.8 Hz, 2H), 2.64 (brs, 4H), 2.49 (brs, 4H), 2.30 (s, 3H), 2.25 (s, 3H), 1.84-1.75 (m, 1H), 1.72-1.62 (m, 1H), 0.91 (t, J=7.4 Hz, 3H); LCMS C24H34N4OS method (B) Rt=4.535 min, ESI+ m/z=427.3 (M+H).
1-(4-Methyl-4′-(2-(4-methylpiperazin-1-yl)-2-oxoethoxy)-[1,1′-biphenyl]-3-yl)-1-propylthiourea OR0597-2 (method A, 47%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 7.49 (dd, J=8.0, 1.9 Hz, 2H), 7.47 (d, J=8.8 Hz, 2H), 7.39 (s, 1H), 7.37 (d, J=8.0 Hz, 1H), 7.30 (d, J=1.9 Hz, 1H), 7.02 (d, J=8.8 Hz, 2H), 5.50 (brs, 2H), 4.73 (s, 2H), 3.67-3.56 (m, 4H), 2.44-2.36 (m, 4H), 2.29 (s, 3H), 2.25 (s, 3H), 1.86-1.74 (m, 1H), 1.74-1.62 (m, 1H), 0.91 (t, J=7.4 Hz, 3H); LCMS C24H32N4O2S method (B) Rt=4.498 min, ESI+ m/z=441.3 (M+H). 1-(2-Methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethyl) pyridin-3-yl)phenyl)-1-propylthiourea OR0599-2 (method A, 81%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 8.69 (d, J=2.0 Hz, 1H), 7.74 (dd, J=8.0, 2.0 Hz, 1H), 7.52 (dd, J=8.0, 2.0 Hz, 1H), 7.43 (d, J=8.0 Hz, 1H), 7.32 (d, J=2.0 Hz, 1H), 7.27 (d, J=8.0 Hz, 1H), 5.53 (brs, 2H), 4.48-4.41 (m, 1H), 3.69-3.61 (m, 1H), 3.05-3.01 (m, 2H), 2.81-2.77 (m, 2H), 2.59 (brs, 4H), 2.48 (brs, 4H), 2.29 (s, 3H), 2.28 (s, 3H), 1.82-1.63 (m, 2H), 0.91 (t, J=7.4 Hz, 3H); LCMS C23H33N5S method (B) Rt=3.891 min, ESI+ m/z=412.3 (M+H).
General Procedure for the Synthesis of N-([1,1′-biaryl]-3-ylcarbamothioyl)acetamide Derivatives. As previously described for N-((3-nitroaryl)carbamothioyl)acetamide derivatives.
N-((4-Methyl-4′-((4-methylpiperazin-1-yl)methyl)-[1,1′-biphenyl]-3-yl)carbamothioyl)acetamide OR0274-3 (method A, 67%) as a light brown solid, engaged in the next step without further purification.
N-((4-Methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)carbamothioyl)acetamide OR0325-3 (method A, 83%) as a light brown solid used in the next step without further purification.
N-((4′-Hydroxy-4-methyl-[1,1′-biphenyl]-3-yl)carbamothioyl)acetamide OR0320-3 (72%) as a white solid. 1H NMR (400 MHz, MeOD) δ 7.88 (d, J=1.9 Hz, 1H), 7.45 (d, J=8.7 Hz, 2H), 7.39 (dd, J=7.9, 1.9 Hz, 1H), 7.28 (d, J=7.9 Hz, 1H), 6.83 (d, J=8.7 Hz, 2H), 2.27 (s, 3H), 2.18 (s, 3H); LCMS C16H16N2O2S method (A) Rt=6.114 min, ESI+ m/z=301.0 (M+H).
N-((4′-Hydroxy-3′-methoxy-4-methyl-[1,1′-biphenyl]-3-yl)carbamothioyl)acetamide OR0321-3 (method A, 88%), as a light brown solid. 1H NMR (400 MHz, MeOD) δ 7.90 (d, J=1.8 Hz, 1H), 7.41 (dd, J=7.9, 1.8 Hz, 1H), 7.29 (d, J=7.9 Hz, 1H), 7.16 (d, J=2.0 Hz, 1H), 7.06 (dd, J=8.2, 2.0 Hz, 1H), 6.85 (d, J=8.2 Hz, 1H), 3.91 (s, 3H), 2.28 (s, 3H), 2.19 (s, 3H); LCMS C17H18N2O3S method (A) Rt=6.215 min, ESI+ m/z=331.1 (M+H).
General Procedure for the Synthesis of N-([1,1′-biaryl]-3-ylcarbamothioyl)benzamide derivatives. Under argon, at 0° C., to a solution of appropriate [1,1′-biaryl]-3-N-alkylamine (1.0 mmol) in dry acetone (25 mL) was added dropwise benzoyl isothiocyanate (1.1 mmol). The resulting solution was stirred at 0° C. for additional 1 hour and then allowed to warm to room temperature. After stirring overnight, upon complete consumption of the starting material monitored by TLC and LCMS, the solvent was distillated off under reduced pressure. The crude residue of corresponding N-([1,1′-biaryl]-3-ylcarbamothioyl)benzamide was used in the next step without further purification.
N-((4-Methyl-4′-(3-(4-methylpiperazin-1-yl)propyl)-[1,1′-biphenyl]-3-yl)(propyl)carbamothioyl)benzamide OR0598-3 (quantitative) as a pale yellow solid. Rf=0.40 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.57-7.20 (m, 12H), 4.48-4.42 (m, 1H), 3.86-3.80 (m, 1H), 2.72-2.64 (m, 2H), 2.58 (brs, 8H), 2.48-2.40 (m, 2H), 2.35 (s, 3H), 2.33 (s, 3H), 1.91-1.71 (m, 4H), 0.95 (t, J=7.4 Hz, 3H); LCMS C32H40N4OS method (B) Rt=4.647 min, ESI+ m/z=529.3 (M+H).
N-(8-(4-(2-(4-Methylpiperazin-1-yl)ethyl)phenyl)-2,3,4,5-tetrahydro-1H-benzo[b]azepine-1-carbonothioyl)benzamide OR0402-3 (96%) as a pale yellow solid, used in the next step without further purification; LCMS C31H36N4OS method (B) Rt=3.787 min, ESI+ m/z=513.3 (M+H).
N-((4-Methyl-4′-(2-(4-methylpiperazin-1-yl)ethoxy)-[1,1′-biphenyl]-3-yl)(propyl)carbamothioyl)benzamide OR0596-3 (99%) as a pale yellow solid. Rf=0.36 (DCM-MeOH—NH4OH, 90:9:1); LCMS C31H35N4O2S method (B) Rt=4.982 min, ESI+ m/z=531.3 (M+H).
N-((4-Methyl-4′-(2-(4-methylpiperazin-1-yl)-2-oxoethoxy)-[1,1′-biphenyl]-3-yl)(propyl)carbamothioyl)benzamide OR0597-3 (quantitative) as a white solid. Rf=0.42 (DCM-MeOH—NH4OH, 90:9:1); LCMS C31H36N4O3S method (B)R=4.907 min, ESI+ m/z=545.3 (M+H).
N-((2-Methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethyl)pyridin-3-yl)phenyl)(propyl)carbamothioyl)benzamide OR0599-3 (quantitative) as a white solid was used in the next step without further purification. Rf=0.40 (DCM-MeOH—NH4OH, 90:9:1); LCMS C30H37N5OS method (B) Rt=4.461 min, ESI+ m/z=516.3 (M+H).
General Procedure for the Synthesis of [1,1′-biaryl]-3-N-alkylamine Derivatives. To a stirred solution of appropriate [1,1′-biaryl]-3-amine (1.0 mmol) and aldehyde (1.5 mmol) in anhydrous tetrahydrofuran (8 mL) was added dropwise glacial acetic acid (1.5 mmol). After stirring for 2 hours, sodium triacetoxyborohydride (2.7 mmol) was added portionwise. The resulting suspension was stirred at room temperature, upon complete consumption of starting material monitored by TLC and LCMS. The reaction was cautiously quenched with a saturated aqueous solution of Na2CO3 (20 mL) and extracted with DCM (3×50 mL). The combined organic layers were dried over Na2SO4 and the solvent was distillated off under reduced pressure. The residue was purified by flash chromatography to afford corresponding [1,1′-biaryl]-3-N-alkylamine.
4-Methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-N-propyl-[1,1′-biphenyl]-3-amine OR0345-3 (71%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 7.51 (d, J=8.2 Hz, 2H), 7.25 (d, J=8.2 Hz, 2H), 7.10 (d, J=7.6 Hz, 1H), 6.85 (dd, J=7.6, 1.7 Hz, 1H), 6.79 (d, J=1.7 Hz, 1H), 3.19 (t, J=7.1 Hz, 2H), 2.89-2.81 (m, 2H), 2.69-2.62 (m, 2H), 2.51 (brs, 8H), 2.32 (s, 3H), 2.17 (s, 3H), 1.73 (sext, J=7.4 Hz, 2H), 1.03 (t, J=7.4 Hz, 3H); LCMS C23H33N3 method (B) Rt=4.429 min, ESI+ m/z=352.3 (M+H).
4-Methyl-4′-(3-(4-methylpiperazin-1-yl)propyl)-N-propyl-[1,1′-biphenyl]-3-amine OR0598-4 (64%) as a yellow foam. Rf=0.38 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.50 (d, J=8.2 Hz, 2H), 7.23 (d, J=8.2 Hz, 2H), 7.10 (d, J=7.7 Hz, 1H), 6.86 (dd, J=7.7, 1.7 Hz, 1H), 6.80 (d, J=1.7 Hz, 1H), 3.52 (brs, 1H), 3.20 (t, J=7.1 Hz, 2H), 2.69-2.65 (m, 2H), 2.49 (brs, 8H), 2.43-2.39 (m, 2H), 2.31 (s, 3H), 2.17 (s, 3H), 1.86 (quint, J=7.7 Hz, 2H), 1.72 (sext, J=7.4 Hz, 2H), 1.04 (t, J=7.4 Hz, 3H). 13C NMR (100 MHz, CDCl3) δ 146.57, 140.76, 140.30, 139.76, 130.32, 128.62, 127.06, 120.70, 115.46, 108.44, 58.01, 55.10, 53.09, 46.00, 45.81, 33.37, 28.61, 22.79, 17.15, 11.75; LCMS C24H35N3 method (B) Rt=4.475 min, ESI+ m/z=366.3 (M+H).
4-Methyl-4′-(2-(4-methylpiperazin-1-yl)ethoxy)-N-propyl-[1,1′-biphenyl]-3-amine OR0596-4 (63%) as a pale yellow solid. Rf=0.36 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.50 (d, J=8.8 Hz, 2H), 7.08 (d, J=7.8 Hz, 1H), 6.95 (d, J=8.8 Hz, 2H), 6.82 (dd, J=7.8, 1.8 Hz, 1H), 6.77 (d, J=1.8 Hz, 1H), 4.15 (t, J=5.9 Hz, 2H), 3.52 (brs, 1H), 3.19 (t, J=7.1 Hz, 2H), 2.84 (t, J=5.9 Hz, 2H), 2.64 (brs, 4H), 2.50 (brs, 4H), 2.30 (s, 3H), 2.16 (s, 3H), 1.72 (sext, J=7.4 Hz, 2H), 1.04 (t, J=7.4 Hz, 3H); LCMS C23H33N3O method (B) Rt=4.536 min, ESI+ m/z=368.3 (M+H).
2-((4′-Methyl-3′-(propylamino)-[1,1′-biphenyl]-4-yl)oxy)-1-(4-methylpiperazin-1-yl)ethan-1-one OR0597-4 (82%) as a colorless oil. 1H NMR (400 MHz, CDCl3) δ 7.52 (d, J=8.8 Hz, 2H), 7.09 (d, J=7.6 Hz, 1H), 6.99 (d, J=8.8 Hz, 2H), 6.81 (dd, J=7.6, 1.7 Hz, 1H), 6.76 (d, J=1.7 Hz, 1H), 4.72 (s, 2H), 3.71-3.59 (m, 4H), 3.19 (t, J=7.4 Hz, 2H), 2.46-2.35 (m, 4H), 2.30 (s, 3H), 2.16 (s, 3H), 1.72 (sext, J=7.4 Hz, 2H), 1.04 (t, J=7.4 Hz, 3H); LCMS C23H31N3O2 method (B) Rt=4.433 min, ESI+ m/z=382.3 (M+H).
2-Methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethyl)pyridin-3-yl)-N-propylaniline OR0599-4 (71%) as a colorless oil. Rf=0.38 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 8.74 (d, J=1.8 Hz, 1H), 7.77 (dd, J=8.0, 2.4 Hz, 1H), 7.22 (d, J=8.0 Hz, 1H), 7.12 (d, J=7.7 Hz, 1H), 6.82 (dd, J=7.7, 1.8 Hz, 1H), 6.75 (d, J=1.8 Hz, 1H), 3.58 (brs, 1H), 3.18-3.17 (m, 2H), 3.02-3.00 (m, 2H), 2.83-2.80 (m, 2H), 2.61 (brs, 4H), 2.49 (brs, 4H), 2.30 (s, 3H), 2.17 (s, 3H), 1.73 (sext, J=7.4 Hz, 2H), 1.03 (t, J=7.4 Hz, 3H); LCMS C22H32N4 method (B) Rt=3.941 min, ESI+ m/z=353.3 (M+H).
General Procedure for the Synthesis of [1,1′-biaryl]-3-amine Derivatives. Under hydrogen atmosphere, a solution of appropriate 3-nitro-1,1′-biaryl (1 mmol) and 10% palladium on charcoal (10% w/w) in a mixture of tetrahydrofuran-ethanol (1:1, 20 mL) was stirred at room temperature until the reaction was complete as indicated by TLC monitoring. The reaction mixture was filtrated through a short pad of celite and rinsed with EtOH. The solvent was distillated off under reduced pressure and the residue was purified by flash chromatography to afford the corresponding [1,1′-biaryl]-3-amine.
4-Methyl-4′-((4-methylpiperazin-1-yl)methyl)-[1,1′-biphenyl]-3-amine OR0274-4 (52%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 7.49 (d, J=8.2 Hz, 2H), 7.35 (d, J=8.2 Hz, 2H), 7.11 (d, J=7.7 Hz, 1H), 6.94 (dd, J=7.7, 1.8 Hz, 1H), 6.90 (d, J=1.7 Hz, 1H), 6.90 (d, J=1.7 Hz, 1H), 3.67 (brs, 2H), 3.54 (s, 2H), 2.49 (s, 8H), 2.30 (s, 3H), 2.21 (s, 3H).
4-Methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-amine OR0325-4 (95%) as a white powder. 1H NMR (400 MHz, CDCl3) δ 7.47 (d, J=8.2 Hz, 2H), 7.24 (d, J=8.2 Hz, 2H), 7.10 (d, J=7.7 Hz, 1H), 6.92 (dd, J=7.7, 1.8 Hz, 1H), 6.89 (d, J=1.8 Hz, 1H), 3.67 (brs, 2H), 2.88-2.80 (m, 2H), 2.68-2.60 (m, 2H), 2.51 (brs, 8H), 2.31 (s, 3H), 2.20 (s, 3H); LC method (C) Rt=2.077 min.
3′-Amino-4′-methyl-[1,1′-biphenyl]-4-oI OR0320-4 (quantitative) as a yellow oil, without further purification. 1H NMR (400 MHz, MeOD) δ 7.38 (d, J=8.7 Hz, 2H), 7.00 (d, J=7.7 Hz, 1H), 6.93 (d, J=1.7 Hz, 1H), 6.82 (dd, J=7.7, 1.7 Hz, 1H), 6.80 (d, J=8.7 Hz, 2H), 2.17 (s, 3H); LCMS C13H13NO method (A) Rt=5.325 min, ESI+ m/z=200.2 (M+H).
3′-Amino-3-methoxy-4′-methyl-[1,1′-biphenyl]-4-oI OR0321-4 (81%) as a white powder. 1H NMR (400 MHz, CDCl3) δ 7.10-7.04 (m, 3H), 6.95 (d, J=8.0 Hz, 1H), 6.89 (dd, J=7.7, 1.8 Hz, 1H), 6.86 (d, J=1.8 Hz, 1H), 5.59 (s, 1H), 3.94 (s, 3H), 3.67 (brs, 2H), 2.20 (s, 3H); LCMS C14H15NO2 method (A) Rt=5.721 min, ESI+ m/z=230.2 (M+H).
4-Methyl-4′-(3-(4-methylpiperazin-1-yl)propyl)-[1,1′-biphenyl]-3-amine OR0598-5 (86%) as a yellow foam. Rf=0.35 (DCM-MeOH—NH4OH, 90:9:1). 1H NMR (400 MHz, CDCl3) δ 7.47 (d, J=8.2 Hz, 2H), 7.22 (d, J=8.2 Hz, 2H), 7.10 (d, J=7.7 Hz, 1H), 6.93 (dd, J=7.7, 1.8 Hz, 1H), 6.89 (d, J=1.8 Hz, 1H), 3.66 (brs, 2H), 2.68-2.63 (m, 2H), 2.48 (brs, 8H), 2.42-2.38 (m, 2H), 2.29 (s, 3H), 2.20 (s, 3H), 1.85 (quint, J=7.7 Hz, 2H); LCMS C21H29N3 method (B) Rt=3.659 min, ESI+ m/z=324.3 (M+H).
4-Methyl-4′-(2-(4-methylpiperazin-1-yl)ethoxy)-[1,1′-biphenyl]-3-amine OR0596-5 (quantitative) as a yellow oil. LCMS C20H27N3O method (B) Rt=3.669 min, ESI+ m/z=326.3 (M+H).
2-((3′-Amino-4′-methyl-[1,1′-biphenyl]-4-yl)oxy)-1-(4-methylpiperazin-1-yl)ethan-1-one OR0597-5 (quantitative) as a pinky solid. LCMS C20H25N3O2 method (B) R=3.586 min, ESI+m/z=340.4 (M+H).
2-Methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethyl)pyridin-3-yl)aniline OR0599-5 (76%) as a white solid, which was used in the next step without further purification. 1H NMR (400 MHz, CDCl3) δ 8.70 (d, J=2.0 Hz, 1H), 7.74 (dd, J=8.0, 2.0 Hz, 1H), 7.21 (d, J=8.0 Hz, 1H), 7.13 (d, J=7.7 Hz, 1H), 6.89 (dd, J=7.7, 2.0 Hz, 1H), 6.85 (d, J=2.0 Hz, 1H), 3.72 (brs, 2H), 3.04-3.02 (m, 2H), 2.83-2.81 (m, 2H), 2.64 (brs, 4H), 2.53 (brs, 4H), 2.32 (s, 3H), 2.20 (s, 3H); LCMS C19H26N4 method (B) Rt=1.680 min, ESI+ m/z=311.2 (M+H).
8-(4-(2-(4-Methylpiperazin-1-yl)ethyl)phenyl)-2,3,4,5-tetrahydro-1H-benzo[b]azepine OR0402-4.
A suspension of 1-methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenethyl)piperazine OR0402-5 (330 mg, 1.0 mmol), 8-bromo-2,3,4,5-tetrahydro-1H-benzo[b]azepine hydrochloride (262 mg, 1.0 mmol) PdCl2(dppf) (109 mg, 0.149 mmol), and K2CO3 (839 mg, 6.07 mmol) in a mixture of 1,4-dioxane-water (5:1, 30 mL) was thoroughly degassed several times under argon fillings. The reaction mixture was heated at 80° C. for 1.5 hours. The solvent was distillated off and the residue was triturated with EtOAc-DCM (1:1, 250 mL) and filtered through a pad of celite. The filtrate was concentrated under reduced pressure, and the residue was purified by flash chromatography, gradient DCM-MeOH—NH4OH (100:0:0 to 90:9:1) to afford 8-(4-(2-(4-methylpiperazin-1-yl)ethyl)phenyl)-2,3,4,5-tetrahydro-1H-benzo[b]azepine OR0402-4 (215 mg, 61%) as a yellow foam. Rf=0.27 (DCM-MeOH—NH4OH, 95:5:0.5); 1H NMR (400 MHz, CDCl3) δ 7.49 (d, J=8.2 Hz, 2H), 7.26 (d, J=8.2 Hz, 2H), 7.16 (d, J=7.7 Hz, 1H), 7.04 (dd, J=7.7, 1.8 Hz, 1H), 6.94 (d, J=1.8 Hz, 1H), 3.88 (brs, 1H), 3.12-3.06 (m, 2H), 2.87-2.83 (m, 2H), 2.82-2.78 (m, 2H), 2.70-2.64 (m, 2H), 2.60 (brs, 8H), 2.35 (s, 3H), 1.85-1.81 (m, 2H), 1.70-1.66 (m, 2H)). 13C NMR (100 MHz, CDCl3) δ 150.79, 139.67, 139.09, 139.05, 132.76, 131.38, 129.11, 127.11, 119.56, 118.02, 60.44, 55.10, 53.03, 49.06, 46.01, 35.85, 33.27, 32.10, 27.10; LCMS C23H31N3 method (B) Rt=2.662 min, ESI+ m/z=350.3 (M+H).
General Procedure for the Synthesis of 3-nitro-1,1′-biaryl Derivatives. Under argon, a suspension of appropriate aryl halide (1.0 mmol), 4,4,5,5-tetramethyl-2-(4-methyl-3-nitrophenyl)-1,3,2-dioxaborolane (264 mg, 1.03 mmol), PdCl2(dppf) (36 mg, 0.05 mmol) and Na2CO3 (212 mg, 2 mmol) in a degassed mixture of 1,4-dioxane-water (5:1, 12 mL) was refluxed for 3 hrs, upon complete consumption of starting material. The solvent was distillated off under reduced pressure and the residue purified by flash chromatography to afford corresponding 3-nitro-1,1′-biaryl.
1-Methyl-4-(4′-methyl-3′-nitro-[1,1′-biphenyl]-4-yl)methyl)piperazine OR0274-5 (74%) as a dark oil. 1H NMR (400 MHz, CDCl3) δ 8.19 (d, J=1.9 Hz, 1H), 7.71 (dd, J=8.0, 1.9 Hz, 1H), 7.54 (d, J=8.2 Hz, 2H), 7.42 (d, J=8.2 Hz, 2H), 7.39 (d, J=8.0 Hz, 1H), 3.56 (s, 2H), 2.63 (s, 3H), 2.48 (brs, 8H), 2.30 (s, 3H); LC method (A) Rt=6.840 min.
1-Methyl-4-(2′(4′-methyl-3′-nitro-[1,1′-biphenyl]-4-yl)ethyl)piperazine OR0325-5 (89%) as a dark oil. 1H NMR (400 MHz, CDCl3) δ 8.18 (d, J=1.9 Hz, 1H), 7.70 (dd, J=7.9, 1.9 Hz, 1H), 7.52 (d, J=8.2 Hz, 2H), 7.39 (d, J=7.9 Hz, 1H), 7.31 (d, J=8.2 Hz, 2H), 2.90-2.82 (m, 2H), 2.68-2.63 (m, 2H), 2.63 (s, 3H), 2.51 (s, 8H), 2.31 (s, 3H); LCMS C20H25N3O2 method (B) Rt=4.532 min. ESI+ m/z=340.2 (M+H).
4′-Methyl-3′-nitro-[1,1′-biphenyl]-4-ol OR0320-5 (73%) as a yellow powder. 1H NMR (400 MHz, CDCl3) δ 8.14 (d, J=2.0 Hz, 1H)), 7.67 (dd, J=7.9, 2.0 Hz, 1H), 7.49 (d, J=8.7 Hz, 2H), 7.37 (d, J=7.9 Hz, 1H), 6.93 (d, J=8.7 Hz, 2H), 4.94 (s, 1H), 2.62 (s, 3H); LCMS C13H11NO3 method (A) Rt=6.516 min, ESI+ m/z=230.1 (M+H).
3-Methoxy-4′-methyl-3′-nitro-[1,1′-biphenyl]-4-ol OR0321-5 (69%) as a yellow powder. 1H NMR (400 MHz, CDCl3) δ 8.14 (d, J=1.6 Hz, 1H), 7.67 (dd, J=7.9, 1.6 Hz, 1H), 7.37 (d, J=7.9 Hz, 1H), 7.11 (dd, J=8.2, 1.9 Hz, 1H), 7.06 (d, J=1.9 Hz, 1H), 7.01 (d, J=8.2 Hz, 1H), 5.71 (s, 1H), 3.98 (s, 3H), 2.62 (s, 3H); LCMS C14H13NO4 method (A) Rt=6.637 min, ESI+m/z=260.1 (M+H).
1-Methyl-4-(3′(4′-methyl-3′-nitro-[1,1′-biphenyl]-4-yl)propyl)piperazine OR0598-6 (73%) as a yellow foam. Rf=0.42 (DCM-MeOH—NH4OH, 90:9:1). 1H NMR (400 MHz, CDCl3) δ 8.18 (d, J=1.9 Hz, 1H), 7.70 (dd, J=7.9, 1.9 Hz, 1H), 7.51 (d, J=8.2 Hz, 2H), 7.38 (d, J=8.0 Hz, 1H), 7.28 (d, J=8.2 Hz, 2H), 2.71-2.66 (m, 2H), 2.62 (s, 3H), 2.47 (brs, 8H), 2.42-2.36 (m, 2H), 2.29 (s, 3H), 1.85 (quint, J=7.7 Hz, 2H); LCMS C21H27N3O2 method (B) R=4.576 min, ESI+ m/z=354.2 (M+H).
1-Methyl-4-(2-((4′-methyl-3′-nitro-[1,1′-biphenyl]-4-yl)oxy)ethyl)piperazine OR0596-6 (72%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 8.15 (d, J=2.0 Hz, 1H), 7.67 (dd, J=8.0, 2.0 Hz, 1H), 7.52 (d, J=8.8 Hz, 2H), 7.36 (d, J=8.0 Hz, 1H), 6.99 (d, J=8.8 Hz, 2H), 4.15 (t, J=5.8 Hz, 2H), 2.85 (t, J=5.8 Hz, 2H), 2.61 (s, 3H), 2.60 (brs, 4H), 2.50 (brs, 4H), 2.30 (s, 3H); LCMS C20H25N3O3 method (B) Rt=4.655 min, ESI+ m/z=356.3 (M+H).
2-((4′-Methyl-3′-nitro-[1,1′-biphenyl]-4-yl)oxy)-1-(4-methylpiperazin-1-yl)ethan-1-one OR0597-6 (72%) as a yellow solid. 1H NMR (400 MHz, CDCl3) δ 8.14 (d, J=1.8 Hz, 1H), 7.66 (dd, J=7.9, 1.8 Hz, 1H), 7.52 (d, J=8.8 Hz, 2H), 7.37 (d, J=7.9 Hz, 1H), 7.04 (d, J=8.8 Hz, 2H), 4.74 (s, 2H), 3.68-3.58 (m, 4H), 2.61 (s, 3H), 2.44-2.37 (m, 4H), 2.30 (s, 3H); LCMS C20H23N3O4 method (B) Rt=4.555 min, ESI+ m/z=370.4 (M+H).
1-Methyl-4-(2-(5-(4-methyl-3-nitrophenyl)pyridin-2-yl)ethyl)piperazine OR0599-6 (79%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 8.75 (d, J=2.4 Hz, 1H), 8.16 (d, J=1.9 Hz, 1H), 7.80 (dd, J=8.1, 2.4 Hz, 1H), 7.70 (dd, J=7.9, 1.9 Hz, 1H), 7.44 (d, J=7.9 Hz, 1H), 7.30 (d, J=8.1 Hz, 1H), 3.06-3.03 (m, 2H), 2.82-2.80 (m, 2H), 2.64 (s, 3H), 2.60 (brs, 4H), 2.48 (brs, 4H), 2.29 (s, 3H); LCMS C19H24N4O2 method (B) Rt=3.942 min, ESI+ m/z=341.2 (M+H).
General Procedure for the Synthesis of 1-(Arylalkyl)-4-methylpiperazine Derivatives. Under argon, to a solution of appropriate arylalkyl methanesulfonate (10 mmol) in acetonitrile (30 mL) were successively added potassium carbonate (4.13 g, 30 mmol) and N-methyl-piperazine (1.40 mL, 15 mmol). The reaction mixture was heated at 60° C., until complete conversion monitored by TLC. The resulting mixture was allowed to cool to room temperature, then poured into H2O (100 mL) and extracted with DCM (3×30 mL). The combined organic layers were washed with brine and dried over Na2SO4. The solvent was distillated off and the residue was purified by flash chromatography, to afford corresponding 1-(arylalkyl)-4-methylpiperazine.
1-(4-Bromophenethyl)-4-methylpiperazine OR0325-6 (68%) as a colorless oil. 1H NMR (400 MHz, CDCl3) δ 7.39 (d, J=8.3 Hz, 2H), 7.07 (d, J=8.3 Hz, 2H), 2.79-2.71 (m, 2H), 2.62-2.54 (m, 2H), 2.53 (brs, 8H), 2.32 (s, 3H). LC method (D) Rt=5.648 min.
1-(3-(4-Bromophenyl)propyl)-4-methylpiperazine OR0598-7 (98%) was used in the next step without further purification. Rf=0.40 (DCM-MeOH—NH4OH, 90:9:1). 1H NMR (400 MHz, CDCl3) δ 7.38 (d, J=8.4 Hz, 2H), 7.05 (d, J=8.4 Hz, 2H), 2.62-2.54 (m, 2H), 2.44 (brs, 8H), 2.38-2.30 (m, 2H), 2.28 (s, 3H), 1.78 (quint, J=7.7 Hz, 2H); LCMS C14H21BrN2 method (B) Rt=1.174 min, ESI+ m/z=297.1 (M+H).
1-(2-(5-Bromopyridin-2-yl)ethyl)-4-methylpiperazine OR0599-7 (81%) was used in the next step without further purification. 1H NMR (400 MHz, CDCl3) δ 8.55 (d, J=2.2 Hz, 1H), 7.69 (dd, J=8.3, 2.2 Hz, 1H), 7.08 (d, J=8.3 Hz, 1H), 2.96-2.88 (m, 2H), 2.76-2.68 (m, 2H), 2.54 (brs, 4H), 2.43 (brs, 4H), 2.27 (s, 3H); LCMS C12H15BrN3 method (B) Rt=1.921 min, ESI+ m/z=284.0 (M+H). 1-(2-(4-Bromophenoxy)ethyl)-4-methylpiperazine OR0596-7 (88%) as a colorless oil. 1H NMR (400 MHz, CDCl3) δ 7.35 (d, J=9.0 Hz, 2H), 6.78 (d, J=9.0 Hz, 2H), 4.06 (t, J=5.8 Hz, 2H), 2.79 (t, J=5.8 Hz, 2H), 2.60 (brs, 4H), 2.47 (brs, 4H), 2.28 (s, 3H); LCMS C13H19BrN2O method (B) Rt=4.004 min. ESI+ m/z=299.1 (M+H).
1-Methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenethyl)piperazine OR0402-5 (65%) as a white solid. Rf=0.40 (EP/EA, 5:5); 1H NMR (400 MHz, CDCl3) δ 7.73 (d, J=8.0 Hz, 2H), 7.21 (d, J=8.0 Hz, 2H), 2.86-2.78 (m, 2H), 2.67-2.59 (m, 2H), 2.61 (brs, 8H), 2.34 (s, 3H), 1.33 (s, 12H); 13C NMR (100 MHz, CDCl3) δ 143.63, 135.09, 128.27, 83.82, 60.11, 55.00, 52.81, 45.89, 33.78, 24.98; LCMS C19H31BN2O2 method (B) Rt=3.392 min, ESI+ m/z=331.3 (M+H).
2-(4-Bromophenoxy)-1-(4-methylpiperazin-1-yl)ethan-1-one OR0597-7.
By peptide coupling procedure, previously described for the synthesis of di-tert-butyl (2-(2-((3-benzamidoaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate derivatives, (88%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 7.37 (d, J=9.1 Hz, 2H), 6.83 (d, J=9.1 Hz, 2H), 4.66 (s, 2H), 3.66-3.60 (m, 2H), 3.59-3.53 (m, 2H), 2.41-2.33 (m, 4H), 2.28 (s, 3H); LCMS C13H17BrN2O2 method (B) Rt=3.939 min, ESI+ m/z=313.1 (M+H).
General Procedure for the Synthesis of 1-(Arylalkyl) Methanesulfonate Derivatives. Under argon, at −5° C., to a solution of appropriate commercially available alcohol (10 mmol) and triethylamine (2.8 mL, 20 mmol) in dry dichloromethane (30 mL) was added dropwise methanesulfonyl chloride (1.15 mL, 14.9 mmol). After addition, the reaction mixture was stirred at −5° C. for 45 min, until complete conversion monitored by TLC, then poured into a saturated NaHCO3 aqueous solution (50 mL) and extracted twice with DCM (20 mL). The combined organic layers were washed with brine and dried over Na2SO4. The solvent was distillated off to afford corresponding arylalkyl methanesulfonate, used in the next step without further purification. 4-Bromophenethyl methanesulfonate OR0325-7 (quantitative) as a white solid. 1H NMR (400 MHz, CDCl3) δ 7.45 (d, J=8.4 Hz, 2H), 7.13 (d, J=8.4 Hz, 2H), 4.39 (t, J=6.8 Hz, 2H), 3.01 (t, J=6.8 Hz, 2H), 2.89 (s, 3H); LCMS C9H11BrO3S method (B) Rt=6.025 min, ESI+m/z=301.0 (M+Na).
3-(4-Bromophenyl)propyl methanesulfonate OR0598-8 (95%) as a white solid. Rf=0.55 (DCM). 1H NMR (400 MHz, CDCl3) δ 7.42 (d, J=8.4 Hz, 2H), 7.07 (d, J=8.4 Hz, 2H), 4.21 (t, J=6.3 Hz, 2H), 2.99 (s, 3H), 2.75-2.67 (m, 2H), 2.10-1.99 (m, 2H); LC method (B) Rt=5.642 min.
2-(4-Bromophenoxy)ethyl methanesulfonate OR0596-8 (quantitative) as a colorless oil. LC method (B) Rt=6.010 min. 2-(5-Bromopyridin-2-yl)ethyl methanesulfonate OR0599-8 (quantitative) as a colorless oil, was used in the next step without further purification. LCMS C8H10BrNO3S method (B) Rt=5.280 min, ESI+ m/z=280.0 (M+H).
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenethyl methanesulfonate OR0402-6 (91%) as a white solid, used in the next step without further purification. Rf=0.30 (PE/EtOAc, 1:1); 1H NMR (400 MHz, CDCl3) δ 7.77 (d, J=7.9 Hz, 2H), 7.24 (d, J=7.9 Hz, 2H), 4.42 (t, J=6.9 Hz, 2H), 3.06 (t, J=6.9 Hz, 2H), 2.83 (s, 3H), 1.34 (s, 12H); LCMS C15H23BO5S method (B) Rt=5.498 min, ESI+ m/z=327.2 (M+H).
4-(4-Aminopyrimidin-2-yl)-N-(3-(arylethynyl)aryl)thiazol-2-amine derivatives were prepared by convergent synthesis, from commercially available 4-amino-2-chloropyrimidine and appropriate bromo-3-nitrobenzene (Scheme 3). The 4-amino-2-chloropyrimidine was protected using Boc2O to afford corresponding bis-carbamate. A Stille cross-coupling reaction with tributyl(1-ethoxyvinyl)tin gives the enol ether which was then turned into the corresponding α-bromoketone using N-bromosuccinimide. Starting from appropriate bromo-3-nitrobenzene, a Sonogashira cross-coupling reaction with appropriate 4-ethynylbenzaldehyde allowed to introduce the key 1,2-diarylethyne scaffold. Reductive amination afforded the corresponding benzylamine, while reduction of the nitro group followed by condensation with acetylisothiocyanate and saponification gave the corresponding thiourea. The thiourea was then engaged in a Hantzsch thiazole synthesis with the α-bromoketone leading to the corresponding thiazole. Finally, deprotection with TFA led to expected 4-(4-aminopyrimidin-2-yl)-N-(3-(arylethynyl)aryl)thiazol-2-amines.
General Procedure for the Synthesis of 4-(4-Aminopyrimidin-2-yl)-N-(3-(arylethynyl)aryl)thiazol-2-amine Derivatives. As previously described for N-(3-((4-(4-aminopyrimidin-2-yl)thiazol-2-yl)amino)-4-aryl)benzamide derivatives.
4-(4-Aminopyrimidin-2-yl)-N-(5-((4-((dimethylamino)methyl)phenyl)ethynyl)-2-methylphenyl)thiazol-2-amine OR0237 (30%) as a white powder. 1H NMR (400 MHz, CDCl3) δ 8.30 (d, J=5.8 Hz, 1H), 7.67 (d, J=1.2 Hz, 1H), 7.60 (s, 1H), 7.50 (d, J=8.1 Hz, 2H), 7.29 (d, J=8.1 Hz, 2H), 7.24 (dd, J=7.8, 1.2 Hz, 1H), 7.22 (d, J=7.8 Hz, 1H), 6.34 (d, J=5.8 Hz, 1H), 4.98 (s, 2H), 3.43 (s, 2H), 2.32 (s, 3H), 2.24 (s, 6H); 13C NMR (100 MHz, CDCl3) δ 166.21, 163.10, 160.53, 156.60, 150.57, 139.37, 138.76, 131.71, 131.45, 130.23, 129.24, 128.02, 123.03, 122.49, 121.96, 111.40, 103.46, 89.63, 88.85, 64.23, 45.49, 18.11; LC method (D) Rt=4.030 min.
4-(4-Aminopyrimidin-2-yl)-N-(2-methyl-5-((4-((4-methylpiperazin-1-yl)methyl)phenyl)ethynyl)phenyl)thiazol-2-amine OR0153 (45%) as a white powder. 1H NMR (400 MHz, CDCl3) δ 8.30 (d, J=5.7 Hz, 1H), 7.67 (d, J=1.2 Hz, 1H), 7.60 (s, 1H), 7.48 (d, J=8.0 Hz, 2H), 7.31 (d, J=8.0 Hz, 2H), 7.24 (dd, J=7.8, 1.2 Hz, 1H), 7.22 (d, J=8.0 Hz), 6.34 (d, J=5.7 Hz, 1H), 4.98 (s, 2H), 3.51 (s, 2H), 2.47 (brs, 8H), 2.32 (s, 3H), 2.29 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 166.37, 163.13, 160.55, 156.55, 150.59, 138.84, 131.65, 131.45, 130.48, 129.24, 128.10, 123.30, 122.45, 121.91, 114.19, 111.32, 103.50, 89.60, 88.81, 62.82, 55.17, 53.10, 46.05, 18.08; LC method (D) Rc=3.865 min.
General Procedure for the Synthesis of di-tert-butyl (2-(2-((3-(arylethynyl)aryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate Derivatives. As previously described for di-tert-butyl (2-(2-((3-nitroaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate derivatives.
Di-tert-butyl (2-(2-((5-((4-((dimethylamino)methyl)phenyl)ethynyl)-2-methylphenyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0237-1 (60%) as a light brown powder. 1H NMR (400 MHz, CDCl3) δ 8.67 (d, J=5.7 Hz, 1H), 7.67 (d, J=1.3 Hz, 1H), 7.53 (d, J=5.1 Hz, 1H), 7.53 (s, 1H), 7.50 (d, J=8.2 Hz, 2H), 7.31 (d, J=8.2 Hz, 2H), 7.26 (dd, J=7.8, 1.3 Hz, 1H), 7.23 (d, J=7.8 Hz, 1H), 3.47 (s, 2H), 2.32 (s, 3H), 2.27 (s, 6H), 1.56 (s, 18H).
Di-tert-butyl (2-(2-((2-methyl-5-((4-((4-methylpiperazin-1-yl)methyl)phenyl)ethynyl)phenyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0153-1(44%) as a light brown powder. 1H NMR (400 MHz, CDCl3) δ 8.68 (d, J=5.7 Hz, 1H), 7.67 (d, J=1.4 Hz, 1H), 7.53 (d, J=5.9 Hz, 1H), 7.53 (s, 1H), 7.48 (d, J=8.2 Hz, 2H), 7.31 (d, J=8.2 Hz, 2H), 7.25 (dd, J=7.9, 1.4 Hz, 1H), 7.22 (d, J=7.9 Hz, 1H), 3.53 (s, 2H), 2.55 (brs, 8H), 2.36 (s, 3H), 2.32 (s, 3H), 1.56 (s, 18H).
General Procedure for the Synthesis of 1-(3-(arylethynyl)aryl)thiourea Derivatives. As previously described for 1-(3-nitroaryl)thiourea derivatives.
1-(5-((4-((Dimethylamino)methyl)phenyl)ethynyl)-2-methylphenyl)thiourea OR0237-2 (72%) as a white powder. 1H NMR (400 MHz, MeOD) δ 7.53-7.47 (m, 3H), 7.40-7.30 (m, 4H), 3.50 (s, 2H), 2.30 (s, 3H), 2.25 (s, 6H).
1-(2-Methyl-5-((4-((4-methylpiperazin-1-yl)methyl)phenyl)ethynyl)phenyl)thiourea OR0153-2 (65%) as a white powder. 1H NMR (400 MHz, MeOD) δ 7.53-7.21 (m, 7H), 3.55 (s, 2H), 2.51 (brs, 8H), 2.28 (s, 6H).
General Procedure for the Synthesis of N-((3-(arylethynyl)aryl)carbamothioyl)acetamide Derivatives. As previously described for N-((3-nitroaryl)carbamothioyl)acetamide derivatives.
N-((5-((4-((Dimethylamino)methyl)phenyl)ethynyl)-2-methylphenyl)carbamothioyl)acetamide OR0237-3 as a light brown solid, engaged in the next step without further purification. 1H NMR (400 MHz, CDCl3) δ 12.01 (s, 1H), 9.10 (s, 1H), 7.86 (d, J=1.3 Hz, 1H), 7.57 (d, J=8.3 Hz, 2H), 7.53 (d, J=8.3 Hz, 2H), 7.39 (dd, J=7.9, 1.3 Hz, 1H), 7.24 (d, J=7.9 Hz, 1H), 4.09 (s, 2H), 2.70 (s, 6H), 2.32 (s, 3H), 2.26 (s, 3H).
N-((2-Methyl-5-((4-((4-methylpiperazin-1-yl)methyl)phenyl)ethynyl)phenyl) carbamothioyl)acetamide OR0153-3 as a light brown solid, engaged in the next step without further purification. 1H NMR (400 MHz, CDCl3) δ 11.99 (s, 1H), 9.07 (s, 1H), 7.83 (d, J=1.3 Hz, 1H), 7.47 (d, J=8.1 Hz, 2H), 7.37 (dd, J=7.8, 1.3 Hz, 1H), 7.28 (d, J=8.1 Hz, 2H), 7.24 (d, J=7.8 Hz, 1H), 3.60 (s, 2H), 2.90 (brs, 8H), 2.66 (s, 3H), 2.31 (s, 3H), 2.25 (s, 3H).
General Procedure for the Synthesis of 3-(arylethynyl)aniline Derivatives. As previously described for di-tert-butyl (2-(2-((3-aminoaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate derivatives.
5-((4-((Dimethylamino)methyl)phenyl)ethynyl)-2-methylaniline OR0237-4 engaged in the next step without further purification. 1H NMR (400 MHz, CDCl3) δ 7.45 (d, J=8.1 Hz, 2H), 7.29 (d, J=8.1 Hz, 2H), 7.01 (d, J=7.7 Hz, 1H), 6.88 (dd, J=7.7, 1.9 Hz, 1H), 6.85 (d, J=1.9 Hz, 1H), 3.62 (brs, 2H), 3.50 (s, 2H), 2.46 (s, 2H), 2.25 (s, 6H), 2.18 (s, 3H).
2-Methyl-5-((4-((4-methylpiperazin-1-yl)methyl)phenyl)ethynyl)aniline OR0153-4 engaged in the next step without further purification. 1H NMR (400 MHz, CDCl3) δ 7.45 (d, J=8.1 Hz, 2H), 7.29 (d, J=8.1 Hz, 2H), 7.01 (d, J=7.7 Hz, 1H), 6.88 (dd, J=7.7, 1.1 Hz, 1H), 6.85 (d, J=1.1 Hz, 1H), 3.62 (brs, 2H), 3.50 (s, 2H), 2.46 (brs, 8H), 2.29 (s, 3H), 2.18 (s, 3H).
General Procedure for the Synthesis of (4-((3-nitroaryl)ethynyl)aryl)methanamine Derivatives. To a solution of appropriate 4-((3-nitroaryl)ethynyl)benzaldehyde (1 mmol) and amine (3 mmol) in a mixture of 1,2-dichloroethane-tetrahydrofuran (10:1, 11 mL) was added few drops of acetic acid. The reaction mixture was stirred 5 min, before portion wise addition of sodium triacetoxyborohydride (424 mg, 2 mmol). The resulting suspension was stirred at room temperature overnight, upon complete consumption of starting material monitored by TLC. The reaction was slowly quenched with saturated aqueous NaHCO3 (10 mL) and extracted with DCM (3×10 mL). The combined organic layers were washed with brine (10 mL) and dried over Na2SO4. The solvent was distillated off under reduced pressure and the residue was purified by flash chromatography to afford corresponding (4-((3-nitroaryl)ethynyl)aryl)methanamine.
N,N-Dimethyl-1-(4-((4-methyl-3-nitrophenyl)ethynyl)phenyl)methanamine OR0237-5 (51%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 8.12 (d, J=1.6 Hz, 1H), 7.61 (dd, J=7.9, 1.6 Hz, 1H), 7.49 (d, J=8.2 Hz, 2H), 7.33-7.31 (m, 3H), 3.44 (s, 2H), 2.61 (s, 3H), 2.25 (s, 6H).
1-Methyl-4-(4-((4-methyl-3-nitrophenyl)ethynyl)benzyl)piperazine OR0153-5 (53%) as a light brown powder. 1H NMR (400 MHz, CDCl3) δ 8.11 (d, J=1.7 Hz, 1H), 7.61 (dd, J=7.9, 1.7 Hz, 1H), 7.48 (d, J=8.2 Hz, 2H), 7.33 (d, J=8.2 Hz, 2H), 7.32 (d, J=7.9 Hz, 1H), 3.52 (s, 2H), 2.61 (s, 3H), 2.49 (brs, 8H), 2.31 (s, 3H).
General Procedure for the Synthesis of 4-((3-nitroaryl)ethynyl)benzaldehyde Derivatives. A solution of appropriate bromo-3-nitrobenzene (1 mmol) and 4-ethynylbenzaldehyde (1.4 mmol) in a mixture of dimethylformamide-triethylamine (1:1, 6 mL) was thoroughly degassed several times under argon fillings. Bis(triphenylphosphine)palladium (II) dichloride (35 mg, 0.05 mmol), copper iodide (10 mg, 0.05 mmol) and triphenylphosphine (26 mg, 0.1 mmol) were successively added. The reaction mixture was heated at 70° C. until complete conversion monitored by TLC. The solvent was distillated off under reduced pressure and the residue was purified by flash chromatography to afford corresponding 4-((3-nitroaryl)ethynyl)benzaldehyde.
4-((4-Methyl-3-nitrophenyl)ethynyl)benzaldehyde OR0153-6 (quantitative) as a light brown powder. 1H NMR (400 MHz, CDCl3) δ 10.03 (s, 1H), 8.15 (d, J=1.6 Hz, 1H), 7.89 (d, J=8.2 Hz, 2H), 7.69 (d, J=8.2 Hz, 2H), 7.65 (dd, J=7.9, 1.6 Hz, 1H), 7.36 (d, J=7.9 Hz, 1H), 2.63 (s, 3H); LC method (A) Rt=7.467 min.
4-(4-Aminopyrimidin-2-yl)-N-(2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)phenoxy)phenyl)thiazol-2-amine and benzyloxyphenyl analogue were prepared by convergent synthesis, in ten steps, from commercially available 4-amino-2-chloropyrimidine and 4-methyl-3-nitrophenol (Scheme 4). The 4-amino-2-chloropyrimidine was protected using Boc2O to afford corresponding bis-carbamate. A Stille cross-coupling reaction with tributyl(1-ethoxyvinyl)tin gave the enol ether which was then turned into the corresponding α-bromoketone using N-bromosuccinimide. Starting from 4-methyl-3-nitrophenol, O-arylation in the Chan-Lam coupling conditions, with (4-formylphenyl)boronic acid allowed to construct the diphenyl ether moiety while Mistunobu reaction with appropriate arylmethanol led to corresponding benzyloxyphenyl derivative. Reductive amination afforded the corresponding amine, while reduction of the nitro group followed by condensation with acetylisothiocyanate and saponification gave the corresponding thiourea. The thiourea was then engaged in a Hantzsch thiazole synthesis with the α-bromoketone leading to the corresponding thiazole. Finally, deprotection with TFA led to expected 4-(4-aminopyrimidin-2-yl)-N-(2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)phenoxy)phenyl)thiazol-2-amine and 4-(4-aminopyrimidin-2-yl)-N-(2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)benzyloxy)phenyl)thiazol-2-amine, respectively.
4-(4-Aminopyrimidin-2-yl)-N-(2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)phenoxy)phenyl)thiazol-2-amine OR0143
As previously described for N-(3-((4-(4-aminopyrimidin-2-yl)thiazol-2-yl)amino)-4-aryl)benzamide derivatives, (62%) as a white powder. 1H NMR (400 MHz, CDCl3) δ 8.29 (d, J=5.7 Hz, 1H), 7.56 (s, 1H), 7.29 (d, J=8.5 Hz, 2H), 7.18 (d, J=2.4 Hz, 1H), 7.16 (d, J=8.3 Hz, 1H), 6.98 (d, J=8.5 Hz, 2H), 6.70 (dd, J=8.3, 2.4 Hz, 1H), 6.32 (d, J=5.7 Hz, 1H), 4.98 (brs, 2H), 3.48 (s, 2H), 2.47 (brs, 8H), 2.28 (s, 3H), 2.26 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.76, 163.07, 160.44, 156.67, 156.47, 156.14, 150.46, 139.61, 133.30, 132.05, 130.67, 123.45, 118.79, 114.33, 111.23, 110.01, 103.42, 62.45, 55.21, 53.07, 46.11, 17.32; LC method (C) Rt=3.476 min.
4-(4-Aminopyrimidin-2-yl)-N-(2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)benzyloxy)phenyl)thiazol-2-amine OR0232
As previously described for N-(3-((4-(4-aminopyrimidin-2-yl)thiazol-2-yl)amino)-4-aryl)benzamide derivatives, (70%) as a white powder. 1H NMR (400 MHz, CDCl3) δ 8.29 (d, J=5.7 Hz, 1H), 7.56 (s, 1H), 7.37 (d, J=8.1 Hz, 2H), 7.33 (d, J=8.1 Hz, 2H), 7.16 (d, J=2.5 Hz, 1H), 7.12 (d, J=8.3 Hz, 1H), 6.69 (dd, J=8.3, 2.5 Hz, 1H), 6.32 (d, J=5.7 Hz, 1H), 5.04 (s, 2H), 5.00 (brs, 2H), 3.51 (s, 2H), 2.46 (brs, 8H), 2.28 (s, 3H), 2.22 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 13C NMR (100 MHz, CDCl3) δ 166.07, 163.09, 160.50, 158.10, 156.47, 150.47, 139.35, 138.16, 135.72, 131.82, 129.52, 127.55, 121.33, 111.08, 110.84, 106.67, 103.39, 70.19, 62.81, 55.20, 53.15, 46.10, 17.14; LC method (C) Rt=3.679 min.
Di-tert-butyl (2-(2-((2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)phenoxy)phenyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0143-1
As previously described for di-tert-butyl (2-(2-((3-nitroaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate derivatives, (48%) as a light brown powder. 1H NMR (400 MHz, CDCl3) δ 8.68 (d, J=5.7 Hz, 1H), 7.52 (d, J=5.7 Hz, 1H), 7.49 (s, 1H), 7.36 (brs, 1H), 7.28 (d, J=8.6 Hz, 2H), 7.20 (d, J=2.4 Hz, 1H), 7.17 (d, J=8.3 Hz, 1H), 6.97 (d, J=8.6 Hz, 2H), 6.70 (dd, J=8.3, 2.4 Hz, 1H), 3.49 (s, 2H), 2.49 (brs, 8H), 2.31 (s, 3H), 2.27 (s, 3H), 1.55 (s, 18H).
Di-tert-butyl (2-(2-((2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)benzyloxy)phenyl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate OR0232-1
As previously described for di-tert-butyl (2-(2-((3-nitroaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate derivatives, (76%) as a light brown powder. 1H NMR (400 MHz, CDCl3) δ 8.69 (d, J=5.7 Hz, 1H), 7.53 (d, J=5.7 Hz, 1H), 7.50 (s, 1H), 7.38 (d, J=8.2 Hz, 2H), 7.33 (d, J=8.2 Hz, 2H), 7.17 (d, J=2.5 Hz, 1H), 7.13 (d, J=8.3 Hz, 1H), 6.69 (dd, J=8.3, 2.5 Hz, 1H), 5.04 (s, 2H), 3.51 (s, 2H), 2.47 (brs, 8H), 2.29 (s, 3H), 2.23 (s, 3H), 1.56 (s, 18H).
1-(2-Methyl-5-(4-((4-methylpiperazin-1-yl)methyl)phenoxy)phenyl)thiourea OR0143-2
As previously described for 1-(3-nitroaryl)thiourea derivatives, (56%) as a white powder. 1H NMR (400 MHz, CDCl3) δ 7.63 (brs, 1H), 7.30 (d, J=8.6 Hz, 2H), 7.24 (d, J=8.3 Hz, 1H), 6.93 (d, J=8.6 Hz, 2H), 6.92 (dd, J=8.3, 2.4 Hz, 1H), 6.84 (d, J=2.4 Hz, 1H), 5.94 (brs, 2H), 3.50 (s, 2H), 2.48 (brs, 8H), 2.29 (s, 3H), 2.27 (s, 3H).
1-(2-Methyl-5-(4-((4-methylpiperazin-1-yl)methyl)benzyloxy)phenyl)thiourea OR0232-2
As previously described for 1-(3-nitroaryl)thiourea derivatives, (70%) as a white powder. 1H NMR (400 MHz, MeOD) δ 7.40 (d, J=8.1 Hz, 2H), 7.34 (d, J=8.1 Hz, 2H), 7.19 (d, J=8.5 Hz, 1H), 6.90 (dd, J=8.5, 2.5 Hz, 1H), 6.85 (d, J=2.5 Hz, 1H), 5.06 (s, 2H), 3.54 (s, 2H), 2.50 (brs, 8H), 2.28 (s, 3H), 2.19 (s, 3H).
N-((2-Methyl-5-(4-((4-methylpiperazin-1-yl)methyl)phenoxy)phenyl)carbamothioyl)acetamide OR0143-3
As previously described for N-((3-nitroaryl)carbamothioyl)acetamide derivatives, engaged in the next step without further purification.
N-((2-Methyl-5-(4-((4-methylpiperazin-1-yl)methyl)benzyloxy)phenyl)carbamothioyl)acetamide OR0232-3
As previously described for N-((3-nitroaryl)carbamothioyl)acetamide derivatives, engaged in the next step without further purification.
2-Methyl-5-(4-((4-methylpiperazin-1-yl)methyl)phenoxy)aniline OR0143-4
As previously described for di-tert-butyl (2-(2-((3-aminoaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamates, (70%) as a colorless oil. 1H NMR (400 MHz, CDCl3) δ 7.24 (d, J=8.6 Hz, 2H), 6.97 (d, J=8.5 Hz, 1H), 6.93 (d, J=8.6 Hz, 2H), 6.36 (dd, J=8.5, 2.4 Hz, 1H), 6.33 (d, J=2.4 Hz, 1H), 3.62 (brs, 2H), 3.47 (s, 2H), 2.46 (brs, 8H), 2.29 (s, 3H), 2.13 (s, 3H).
2-Methyl-5-(4-((4-methylpiperazin-1-yl)methyl)benzyloxy)aniline OR0232-4
As previously described for di-tert-butyl (2-(2-((3-aminoaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate derivatives, (77%) as a colorless oil. 1H NMR (400 MHz, CDCl3) δ 7.36 (d, J=8.2 Hz, 2H), 7.32 (d, J=8.2 Hz, 2H), 6.93 (d, J=8.5 Hz, 1H), 6.35 (dd, J=8.5, 2.5 Hz, 1H), 6.33 (d, J=2.5 Hz, 1H), 4.98 (s, 2H), 3.59 (brs, 2H), 3.50 (s, 2H), 2.46 (brs, 8H), 2.29 (s, 3H), 2.10 (s, 3H).
1-Methyl-4-(4-(4-methyl-3-nitrophenoxy)benzyl)piperazine OR0143-5
As previously described for (4-((3-nitroaryl)ethynyl)aryl)methanamine derivatives, (84%) as a colorless oil. 1H NMR (400 MHz, CDCl3) δ 7.57 (d, J=2.6 Hz, 1H), 7.32 (d, J=8.6 Hz, 2H), 7.27 (d, J=8.4 Hz, 1H), 7.15 (dd, J=8.4, 2.6 Hz, 1H), 6.97 (d, J=8.6 Hz, 2H), 3.50 (s, 2H), 2.55 (s, 3H), 2.48 (brs, 8H), 2.30 (s, 3H).
1-Methyl-4-(4-((4-methyl-3-nitrophenoxy)methyl)benzyl)piperazine OR0232-5
Under argon, at 0° C., to a solution of 4-methyl-3-nitrophenol (230 mg, 1.5 mmol), (4-((4-methylpiperazin-1-yl)methyl)phenyl)methanol (300 mg, 1.36 mmol) and triphenylphosphine (536 mg, 2.04 mmol) in tetrahydrofuran (68 mL), was added dropwise a solution of diisopropyl azodicarboxylate (430 μL, 2.04 mmol) in tetrahydrofuran (35 mL). The resulting mixture was allowed to warm to room temperature and stirred overnight. The solvent was distillated off under reduced pressure and the residue was purified by flash chromatography, eluent DCM-MeOH—NH4OH (99:1:0.1), to afford 1-methyl-4-(4-((4-methyl-3-nitrophenoxy)methyl)benzyl)piperazine OR0232-5 (368 g, 76%) as a colorless oil. 1H NMR (400 MHz, CDCl3) δ 7.59 (d, J=2.7 Hz, 1H), 7.38-7.34 (m, 4H), 7.23 (d, J=8.5 Hz, 1H), 7.12 (dd, J=8.5, 2.7 Hz, 1H), 5.07 (s, 2H), 3.52 (s, 2H), 2.53 (s, 3H), 2.47 (brs, 8H), 2.29 (s, 3H).
4-(4-Methyl-3-nitrophenoxy)benzaldehyde OR0143-6
To a solution of 4-methyl-3-nitrophenol (1 g, 6.5 mmol), (4-formylphenyl)boronic acid (1.96 g, 13.0 mmol) and copper(II) acetate (1.19 g, 6.5 mmol) in dichloromethane (65 mL), charged with activated molecular sieves, was added triethylamine (4.6 mL, 35 mmol). The reaction mixture was vigorously stirred under air atmosphere until complete conversion monitored by LCMS. The solvent was distillated off under reduced pressure and the residue was purified by flash chromatography to afford 4-(4-methyl-3-nitrophenoxy)benzaldehyde OR0143-6 (0.57 g, 34%) as a light yellow powder. 1H NMR (400 MHz, CDCl3) δ 9.96 (s, 1H), 7.90 (d, J=8.6 Hz, 2H), 7.70 (d, J=2.5 Hz, 1H), 7.38 (d, J=8.4 Hz, 1H), 7.25 (dd, J=8.4, 2.5 Hz, 1H), 7.11 (d, J=8.6 Hz, 2H), 2.61 (s, 3H); LCMS C14H11NO4 method (A) Rt=6.953 min, ESI+ m/z=258.1 (M+H).
The condensation of commercially available 4-methyl-3-nitroaniline and methyl 4-((4-methylpiperazin-1-yl)methyl)benzoate was carried out in the presence of trimethylaluminum, to yield the corresponding amide quantitatively. After reduction of the nitro group, the amine was subjected to acetylisothiocyanate, then saponification gave the corresponding thiourea. Ethyl bromopyruvate was engaged in a Hantzsch thiazole synthesis with the thiourea leading to the corresponding thiazole. Finally, tandem addition/cyclization with malonimidamide led to expected N-(3-((4-(4,6-diaminopyrimidin-2-yl)thiazol-2-yl)amino)-4-methylphenyl)-4-((4-methylpiperazin-1-yl)methyl)benzamide dCKi-2.
N-(3-((4-(4,6-Diaminopyrimidin-2-yl)thiazol-2-yl)amino)-4-methylphenyl)-4-((4-methylpiperazin-1-yl)methyl)benzamide dCKi-2
To a suspension of ethyl 2-((2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)benzamido)phenyl)amino)thiazole-4-carboxylate dCKi-2-1 (2.4 g, 4.8 mmol), malonimidamide dihydrochloride (2.5 g, 14.4 mmol) in methanol (50 mL) was added sodium methoxide solution 25 wt. % in MeOH (14.9 mL, 65.2 mmol). The reaction mixture was heated at 70° C. for 2 hours and concentrated under vacuum. The crude product was triturated with THF (3×50 mL) and combined organic layers were dried over Na2SO4. The solvent was distillated off under reduced pressure and the residue was purified by flash chromatography, gradient DCM-MeOH—NH4OH (100:0:0 to 90:9:1) to afford N-(3-((4-(4,6-diaminopyrimidin-2-yl)thiazol-2-yl)amino)-4-methylphenyl)-4-((4-methylpiperazin-1-yl)methyl)benzamide dCKi-2 (940 mg, 37%) as a light brown powder. Rf=0.10 (DCM-MeOH—NH4OH, 92:7:1); 1H NMR (300 MHz, DMSO-d6) δ 10.22 (brs, 1H), 9.37 (brs, 1H), 8.84 (d, J=1.7 Hz, 1H), 7.90 (d, J=8.2 Hz, 2H), 7.51 (dd, J=8.3, 1.7 Hz, 1H), 7.42 (d, J=8.2 Hz, 2H), 7.31 (s, 1H), 7.20 (d, J=8.3 Hz, 1H), 6.10 (brs, 4H), 5.34 (s, 1H), 3.52 (s, 2H), 2.48-2.30 (m, 8H), 2.22 (s, 3H), 2.19 (s, 3H); 13C NMR (75 MHz, DMSO-d6) δ 165.56, 165.27, 163.66, 159.29, 151.32, 142.01, 139.60, 137.78, 133.60, 130.66, 128.59, 127.59, 125.97, 116.39, 114.39, 109.48, 81.27, 61.45, 54.46, 52.23, 45.38, 17.41; LCMS C2H31N9OS method (A) Rt=6.085 min, ESI+ m/z=530.2 (M+H).
Ethyl 2-((2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)benzamido)phenyl)amino)thiazole-4-carboxylate dCKi-2-1 To a stirred solution of dCKi-2-2 (4.0 g, 10.0 mmol) in ethanol (100 mL) was added ethyl bromopyruvate (1.5 mL, 12.0 mmol). The resulting mixture was refluxed for 3 hrs and then concentrated under reduced pressure. The residue was purified by flash chromatography, gradient DCM-MeOH (100:0 to 80:20) to afford ethyl 2-((2-methyl-5-(4-((4-methylpiperazin-1-yl)methyl)benzamido)phenyl)amino)thiazole-4-carboxylate dCKi-2-1 (4.2 g, 85%) as a light brown solid. Rf=0.32 (DCM-MeOH, 9:1); 1H NMR (250 MHz, MeOD) δ 8.21 (d, J=2.1 Hz, 1H), 7.93 (d, J=8.2 Hz, 2H), 7.64 (s, 1H), 7.58 (dd, J=8.3, 2.1 Hz, 1H), 7.51 (d, J=8.2 Hz, 2H), 7.23 (d, J=8.3 Hz, 1H), 4.34 (q, J=7.1 Hz, 2H), 3.72 (s, 2H), 2.47-3.10 (m, 4H), 2.97-2.60 (m, 4H), 2.84 (s, 3H), 2.30 (s, 3H), 1.37 (t, J=7.1 Hz, 3H); 13C NMR (63 MHz, MeOD) δ 168.32, 168.21, 163.33, 163.33, 143.77, 142.36, 140.26, 138.70, 135.52, 132.06, 130.31, 128.87, 127.00, 119.17, 118.19, 115.40, 62.29, 62.19, 54.92, 51.12, 50.02, 49.68, 49.34, 49.00, 48.66, 48.32, 47.98, 43.74, 17.60, 14.58; LCMS C26H31N5O3S method (A) R=6.325 min, ESI+m/z=494.0 (M+H).
N-[3-(Carbamothioylamino)-4-methyl-phenyl]-4-[(4-methylpiperazin-1-yl)methyl]benzamide dCKi-2-2
As previously described for the synthesis of 1-(3-nitroaryl)thiourea derivatives, (94%) as a light brown solid, which was used directly in the next step without further purification. Rf=0.04 (DCM-MeOH 90:10); 1H NMR (250 MHz, DMSO-d6) δ 10.17 (brs, 1H), 9.32 (brs, 1H), 7.89 (d, J=8.1 Hz, 2H), 7.66 (d, J=2.0 Hz, 1H), 7.62 (dd, J=8.3, 2.0 Hz, 1H), 7.43 (d, J=8.1 Hz, 2H), 7.20 (d, J=8.3 Hz, 1H), 3.52 (s, 2H), 2.35 (brs, 8H), 2.14 (s, 6H); 13C NMR (63 MHz, DMSO-d6) δ 181.46, 165.27, 142.23, 137.59, 137.00, 133.61, 130.43, 129.65, 128.65, 127.58, 119.25, 118.54, 61.62, 54.72, 52.60, 45.76, 17.13; LCMS C21H27N5O2S method (A) Rt=6.259 min, ESI+ m/z=398.0 (M+H).
N-[3-(Acetylcarbamothioylamino)-4-methyl-phenyl]-4-[(4-methylpiperazin-1-yl)methyl]benzamide dCKi-2-3
As previously described for the synthesis of N-((3-nitroaryl)carbamothioyl)acetamide derivatives, (59%) as a light brown solid, which was used directly in the next step without further purification. Rf=0.12 (DCM-MeOH 90:10); 1H NMR (250 MHz, DMSO-d6) δ 12.13 (brs, 1H), 11.52 (brs, 1H), 10.27 (brs, 1H), 7.99 (d, J=1.7 Hz, 1H), 7.94 (d, J=8.2 Hz, 2H), 7.62 (dd, J=8.1, 1.7 Hz, 1H), 7.47 (d, J=8.2 Hz, 2H), 7.25 (d, J=8.1 Hz, 1H), 3.65 (s, 2H), 3.40 (brs, 4H), 3.10 (brs, 4H), 2.70 (s, 3H), 2.17 (s, 6H); 13C NMR (63 MHz, DMSO-d6) δ 179.75, 172.77, 165.15, 140.92, 137.30, 136.65, 133.84, 130.30, 128.87, 128.63, 127.74, 119.11, 118.48, 60.39, 52.75, 49.41, 42.35, 23.79, 17.13; LCMS C23H29N5O2S method (A) Rt=6.074 min, ESI+ m/z=440.0 (M+H).
N-(3-Amino-4-methylphenyl)-4-((4-methylpiperazin-1-yl)methyl)benzamide dCKi-2-4
To a solution of dCKi-2-5 (9.3 g, 25.3 mmol) in a mixture of ethyl acetate and acetic acid (2:1, 300 mL), zinc dust (24.4 g, 25.3 mmol) was added carefully. The resulting mixture was stirred at room temperature for 3 hrs, then filtered through Celite pad and washed with EtOAc (200 mL). The filtrate was concentrated under reduced pressure and the crude residue was successively dissolved in water, then Na2CO3 was added until pH=9, and extracted with EtOAc. The combined organic layer were dried over Na2SO4, and the solvent was distillated off under reduced pressure to afford N-(3-amino-4-methylphenyl)-4-((4-methylpiperazin-1-yl)methyl)benzamide dCKi-2-4 (8.5 g, quantitative) as a light brown solid, used in the next step without further purification. Rf=0.11 (DCM-MeOH—NH4OH 92:7:1); 1H NMR (250 MHz, DMSO-d6, δ) 9.86 (brs, 1H), 7.86 (d, J=8.0 Hz, 2H), 7.40 (d, J=8.0 Hz, 2H), 7.11 (d, J=1.4 Hz, 1H), 6.91-6.73 (m, 2H), 4.83 (brs, 2H), 3.52 (s, 2H), 2.47-2.25 (brs, 8H), 2.19 (s, 3H), 2.01 (s, 3H); LCMS C20H26N4O method (A) Rt=4.346 min, ESI+ m/z=339.0 (M+H).
N-(4-Methyl-3-nitrophenyl)-4-((4-methylpiperazin-1-yl)methyl)benzamide dCKi-2-5
Under argon, at 0° C., to a stirred solution of 4-methyl-3-nitroaniline (4.2 g, 27.6 mmol) and methyl 4-((4-methylpiperazin-1-yl)methyl)benzoate (6.3 g, 25.3 mmol) in a mixture of toluene-tetrahydrofuran (3:1, 320 mL) was added dropwise a 2M solution of trimethylaluminium in toluene (40 mL, 80 mmol). The resulting mixture was allowed to warm to room temperature and heated to 50° C. for 6 hrs. The mixture was poured carefully onto ice (200 g) and quenched by the addition of 10% NaOH aqueous solution (150 mL). After stirring for 30 min, the aqueous phase was extracted with DCM (3×200 mL). The combined organic layers were concentrated to dryness under reduced pressure to afford N-(4-methyl-3-nitrophenyl)-4-((4-methylpiperazin-1-yl)methyl)benzamide dCKi-2-5 (9.3 g, quantitative) as a light brown solid, used in the next step without further purification. Rf=0.27 (DCM-MeOH—NH4OH 92:7:1); 1H NMR (250 MHz, CDCl3) δ 8.25 (d, J=2.3 Hz, 1H), 8.04 (brs, 1H), 7.91 (dd, J=8.3, 2.3 Hz, 2H), 7.83 (d, J=8.2 Hz, 2H), 7.47 (d, J=8.2 Hz, 2H), 7.34 (d, J=8.3 Hz, 1H), 3.58 (s, 2H), 2.59 (s, 3H), 2.70-2.25 (m, 8H), 2.30 (s, 3H); 13C NMR (63 MHz, CDCl3) δ 166.03, 149.07, 143.32, 137.06, 133.33, 132.91, 129.49, 129.33, 127.25, 124.93, 116.28, 62.56, 55.18, 53.22, 46.14, 20.13; LCMS C20H24N4O3 method (A) Rt=6.479 min, ESI+ m/z=369.0 (M+H).
N-([1,1′-biaryl]-3-yl)-4-(4,6-diaminopyrimidin-2-yl)thiazol-2-amine derivatives were prepared in seven steps (Scheme 6). Starting from appropriate bromo-3-nitrobenzene, a Suzuki cross-coupling reaction with arylboronic acid derivatives allowed introducing the key biphenyl scaffold. Reduction of the nitro group successively followed by alkylation, then condensation with benzoyl isothiocyanate and saponification gave the corresponding thiourea. The ethyl bromopyruvate was engaged in a Hantzsch thiazole synthesis with the thiourea leading to the corresponding thiazole. Finally, tandem addition/cyclization with malonimidamide led to expected structurally diverse N-([1,1′-biaryl]-3-yl)-4-(4,6-diaminopyrimidin-2-yl)thiazol-2-amine.
Alternatively, N-([1,1′-biaryl]-3-yl)-4-(4,6-diaminopyrimidin-2-yl)thiazol-2-amine derivatives and analogues, were prepared by convergent approach from commercially available aminohalogenobenzene (Scheme 7). For instance, starting from 5-bromo- or 5-iodo-2-methylaniline, condensation with benzoyl isothiocyanate and saponification gave the corresponding thiourea. Ethyl bromopyruvate was engaged in a Hantzsch thiazole synthesis with the thiourea leading to the corresponding thiazole. The thiazole derivative was then turned into expected N-([1,1′-biaryl]-3-yl)-4-(4,6-diaminopyrimidin-2-yl)thiazol-2-amine by Suzuki cross-coupling reaction via either arylboronate or halogenoaryl precursors.
General Procedure for the Synthesis of 2-[2-(biaryl-3-ylamino)-thiazol-4-yl]-pyrimidine-4,6-diamine hydrochloride salt. To a solution of appropriate 2-[2-(biaryl-3-ylamino)-thiazol-4-yl]-pyrimidine-4,6-diamine (1.5 mmol) in a mixture of methanol-dichloromethane (5:2, 35 mL) was added dropwise a 1N hydrogen chloride solution in diethyl ether (3.2 mL, 3.2 mmol). The resulting solution was stirred at room temperature for 30 minutes and concentrated under reduced pressure until few volumes. The residue was crystallized from Et2O, under vigorous stirring, to afford expected 2-[2-(biaryl-3-ylamino)-thiazol-4-yl]-pyrimidine-4,6-diamine hydrochloride salt.
2-(2-((4-Methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-2′-(trifluoromethyl)-[1,1′-biphenyl]-3-yl)(propyl)amino) thiazol-4-yl)pyrimidine-4,6-diamine hydrochloride salt OR0642 (93%) as a light yellow powder. H NMR (400 MHz, MeOD) δ 8.17 (s, 1H), 8.15 (dd, J=8.1, 1.7 Hz, 1H), 7.74 (d, J=8.0 Hz, 1H), 7.69 (s, 1H), 7.57 (d, J=8.0 Hz, 1H), 7.41 (dd, J=7.9, 1.6 Hz, 1H), 7.34 (s, 1H), 5.67 (s, 1H), 4.28 (brs, 1H), 4.01 (brs, 2H), 3.85 (brs, 1H), 3.61 (brs, 2H), 3.30 (brs, 2H), 2.93 (brs, 2H), 2.92 (s, 3H), 2.34 (s, 3H), 1.72 (sext, J=7.4 Hz, 2H), 1.02 (t, J=7.4 Hz, 3H). 13C NMR (100 MHz, MeOD) δ 172.73, 152.34, 146.43, 144.00, 143.15, 139.26, 138.98, 136.95, 135.12, 133.51, 132.38, 130.94, 130.62, 126.74, 126.04, 123.32, 115.99, 80.46, 54.70, 53.95, 44.53, 43.49, 22.09, 17.51, 11.75.
General Procedure for the Synthesis of 2-[2-(biaryl-3-ylamino)-thiazol-4-yl]-pyrimidine-4,6-diamine and analogues. Method (A): as previously described for synthesis of dCKi-2, briefly to a suspension of ethyl 2-([1,1′-aryl]-3-yl-amino)thiazole-4-carboxylate derivative or analogue (0.76 mmol), malonimidamide dihydrochloride (225 mg, 1.28 mmol) in methanol (8 mL) was added sodium methoxide solution 25 wt. % in MeOH (1.56 mL, 6.80 mmol). The reaction mixture was heated at 70° C. for 4 hours and concentrated under vacuum. The crude product was triturated with THF (3×50 mL) and combined organic layers were dried over Na2SO4. The solvent was distillated off under reduced pressure and the residue was purified by flash chromatography to afford the corresponding 2-[2-(biaryl-3-ylamino)-thiazol-4-yl]-pyrimidine-4,6-diamine or analogue. Method (B): under argon, a suspension of appropriate 2-(2-(5-halogenophenylxamino)thiazol-4-yl)pyrimidine-4,6-diamine (0.214 mmol), arylboronate or arylboronic acid (0.286 mmol), PdCl2(dppf) (21 mg, 0.028 mmol) and K2CO3 (103 mg, 0.745 mmol) in a degassed mixture of 1,4-dioxane-water (5:1, 12 mL) was heated at 80° C. for 1.5 hrs, upon complete consumption of starting material. The solvent was distillated off under reduced pressure and the residue purified by flash chromatography to afford corresponding 2-[2-(biaryl-3-ylamino)-thiazol-4-yl]-pyrimidine-4,6-diamine. Method (C): from appropriate tert-butyl(biaryl-3-yl-[4-(4,6-diamino-pyrimidin-2-yl)-thiazol-2-yl]-aminoalkyl)carbamate, as previously described for the synthesis of N-(3-((4-(4-aminopyrimidin-2-yl)thiazol-2-yl)amino)-4-aryl)benzamide derivatives.
2-(2-((4-Methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0600 (method A, 25%) as a light brown solid. Rf=0.25 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.52 (dd, J=8.2, 2.0 Hz, 1H), 7.50 (d, J=8.4 Hz, 2H), 7.45 (d, J=2.0 Hz, 1H), 7.38 (d, J=8.2 Hz, 1H), 7.31 (s, 1H), 7.27 (d, J=8.4 Hz, 2H), 5.42 (s, 1H), 4.71 (brs, 4H), 4.01 (brs, 2H), 2.86-2.81 (m, 2H), 2.66-2.61 (m, 2H), 2.50 (brs, 8H), 2.30 (s, 3H), 2.26 (s, 3H), 1.66 (sext, J=7.4 Hz, 2H), 0.92 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 171.22, 163.93, 160.87, 151.29, 143.34, 140.77, 139.91, 137.91, 136.29, 132.50, 129.38, 128.02, 127.00, 126.96, 110.80, 83.01, 60.52, 55.27, 53.62, 53.30, 46.20, 33.35, 21.33, 17.49, 11.42; LCMS C30H38N8S method (B) Rc=4.243 min, ESI+ m/z=543.3 (M+H).
2-(2-((4-Methyl-4′-(3-(4-methylpiperazin-1-yl)propyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0601 (method A, 40%) as a light yellow solid. Rf=0.25 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.52 (dd, J=8.0, 1.8 Hz, 1H), 7.49 (d, J=8.1 Hz, 2H), 7.45 (d, J=1.8 Hz, 1H), 7.38 (d, J=8.0 Hz, 1H), 7.31 (s, 1H), 7.25 (d, J=8.1 Hz, 2H), 5.42 (s, 1H), 4.82 (brs, 4H), 4.01 (brs, 2H), 2.66 (t, J=7.7 Hz, 2H), 2.51 (brs, 8H), 2.44-2.38 (m, 2H), 2.31 (s, 3H), 2.25 (s, 3H), 1.85 (quint, J=7.7 Hz, 2H), 1.66 (sext, J=7.5 Hz, 2H), 0.92 (t, J=7.5 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 171.19, 163.80, 160.57, 151.07, 143.31, 141.64, 140.80, 137.61, 136.18, 132.48, 129.06, 127.93, 126.92, 110.87, 82.95, 57.95, 55.10, 53.67, 53.07, 46.02, 33.41, 28.61, 21.32, 17.49, 11.41. 1; LCMS C31H40N8S method (B) Rc=4.279 min, ESI+ m/z=557.3 (M+H).
2-(2-((2-Methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethyl)pyridin-3-yl)phenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0602 (method A, 34%) as a white powder. Rf=0.25 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 8.74 (d, J=2.0 Hz, 1H), 7.77 (dd, J=8.1, 2.0 Hz, 1H), 7.51 (dd, J=7.9, 1.8 Hz, 1H), 7.44 (d, J=1.8 Hz, 1H), 7.43 (d, J=7.9 Hz, 1H), 7.32 (s, 1H), 7.25 (d, J=8.1 Hz, 1H), 5.44 (s, 1H), 4.71 (brs, 4H), 4.01 (brs, 2H), 3.05-3.00 (m, 2H), 2.82-2.77 (m, 2H), 2.60 (brs, 4H), 2.48 (brs, 4H), 2.29 (s, 3H), 2.28 (s, 3H), 1.66 (sext, J=7.4 Hz, 2H), 0.93 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 171.00, 163.92, 160.82, 159.60, 151.33, 147.56, 143.64, 137.64, 137.33, 134.71, 133.18, 132.87, 128.11, 126.94, 123.29, 110.86, 83.02, 58.44, 55.26, 53.65, 53.19, 46.17, 35.53, 21.36, 17.57, 11.41; LCMS C29H37N9S method (B) Rt=3.952 min, ESI+ m/z=544.3 (M+H).
2-(2-(iso-Butyl(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0603 (method A, 23%) as a light brown powder. Rf=0.5 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.52 (dd, J=7.6, 1.9 Hz, 1H), 7.50 (d, J=1.9 Hz, 1H), 7.49 (d, J=8.2 Hz, 2H), 7.38 (d, J=7.6 Hz, 1H), 7.32 (s, 1H), 7.28 (d, J=8.2 Hz, 2H), 5.41 (s, 1H), 4.70 (brs, 4H), 3.84 (brs, 2H), 2.88-2.80 (m, 2H), 2.67-2.61 (m, 2H), 2.56 (brs, 8H), 2.30 (s, 3H), 2.26 (s, 3H), 2.04 (sept, J=6.8 Hz, 1H), 1.00 (d, J=6.8 Hz, 6H). 13C NMR (100 MHz, CDCl3) δ 171.69, 163.92, 160.89, 151.35, 144.07, 140.59, 139.89, 138.00, 135.94, 132.71, 129.39, 127.87, 127.03, 126.81, 111.00, 83.02, 60.53, 59.79, 55.27, 53.30, 46.19, 33.35, 27.43, 20.78, 17.73; LCMS C31H40N8S method (B) Rt=4.368 min, ESI+ m/z=557.2 (M+H).
2-(2-((Cyclopropylmethyl)(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0604 (method A, 8%) as a light brown powder. Rf=0.5 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.53 (dd, J=8.7, 1.9 Hz, 1H), 7.52 (d, J=1.9 Hz, 1H), 7.50 (d, J=8.2 Hz, 2H), 7.38 (d, J=8.7 Hz, 1H), 7.31 (s, 1H), 7.28 (d, J=8.2 Hz, 2H), 5.44 (s, 1H), 4.68 (brs, 4H), 3.96 (brs, 2H), 2.89-2.80 (m, 2H), 2.68-2.60 (m, 2H), 2.53 (brs, 8H), 2.32 (s, 3H), 2.29 (s, 3H), 1.17-1.05 (m, 1H), 0.44-0.37 (m, 2H), 0.19-0.11 (m, 2H); 13C NMR (100 MHz, CDCl3) δ 171.25, 163.90, 160.90, 151.20, 143.55, 140.63, 139.80, 138.01, 136.48, 132.27, 129.37, 128.44, 126.97, 110.95, 83.01, 60.48, 56.51, 55.21, 53.19, 46.12, 33.32, 17.48, 9.88, 3.97; LCMS C31H38N8S method (B) Rt=4.235 min, ESI+ m/z=555.3 (M+H).
2-(2-(iso-Pentyl(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0605 (method A, 28%) as a light yellow solid. Rf=0.5 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.52 (dd, J=7.9, 1.8 Hz, 1H), 7.50 (d, J=8.2 Hz, 2H), 7.43 (d, J=1.8 Hz, 1H), 7.38 (d, J=7.9 Hz, 1H), 7.33 (s, 1H), 7.27 (d, J=8.2 Hz, 2H), 5.45 (s, 1H), 4.85 (brs, 4H), 4.07 (brs, 2H), 2.90-2.82 (m, 2H), 2.73-2.66 (m, 2H), 2.65 (brs, 8H), 2.39 (s, 3H), 2.25 (s, 3H), 1.71-1.59 (m, 1H), 1.50 (q, J=7.3 Hz, 2H), 0.90 (d, J=6.6 Hz, 6H); 13C NMR (100 MHz, CDCl3) δ 171.26, 163.56, 143.30, 140.69, 139.52, 138.03, 136.34, 132.51, 129.38, 127.90, 127.05, 126.97, 111.15, 82.88, 60.12, 54.82, 52.55, 50.66, 45.67, 36.64, 33.12, 26.43, 22.83, 17.54; LCMS C32H40N8S method (B) Rt=4.452 min, ESI+ m/z=571.4 (M+H).
2-(2-(Butyl(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0606 (method A, 19%) as a light brown solid. Rf=0.5 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.53 (dd, J=8.0, 1.7 Hz, 1H), 7.50 (d, J=8.2 Hz, 2H), 7.44 (d, J=1.7 Hz, 1H), 7.38 (d, J=8.0 Hz, 1H), 7.32 (s, 1H), 7.27 (d, J=8.2 Hz, 2H), 5.45 (s, 1H), 4.82 (brs, 4H), 4.05 (brs, 2H), 2.87-2.83 (m, 2H), 2.69-2.65 (m, 2H), 2.57 (brs, 8H), 2.37 (s, 3H), 2.25 (s, 3H), 1.60-1.58 (m, 2H), 1.38-1.35 (m, 2H), 0.89 (t, J=7.3 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 171.28, 163.65, 150.79, 143.30, 140.73, 139.64, 137.99, 136.32, 132.52, 129.38, 127.98, 127.03, 126.98, 111.06, 82.91, 60.24, 52.78, 51.86, 45.84, 33.20, 30.17, 29.84, 20.34, 17.51, 14.12; LCMS C31H40N8S method (B) Rt=4.394 min, ESI+ m/z=557.3 (M+H).
2-(2-((4-Methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0607 (method B, 76%) as a light brown solid. Rf=0.45 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 7.89 (d, J=8.7 Hz, 2H), 7.85 (d, J=8.7 Hz, 2H), 7.71 (dd, J=8.0, 1.8 Hz, 1H), 7.66 (d, J=1.8 Hz, 1H), 7.54 (d, J=8.0 Hz, 1H), 7.25 (s, 1H), 5.54 (s, 1H), 4.01 (brs, 2H), 3.08-3.04 (m, 4H), 2.53-2.49 (m, 4H), 2.31 (s, 3H), 2.26 (s, 3H), 1.74 (sext, J=7.4 Hz, 2H), 0.99 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 171.99, 165.62, 161.25, 152.08, 145.86, 145.04, 140.31, 139.04, 135.59, 134.14, 129.71, 129.44, 128.58, 128.51, 111.01, 82.96, 55.09, 54.87, 46.95, 45.75, 22.31, 17.61, 11.61; LCMS C28H34N8O2S2 method (B) Rt=4.496 min, ESI+ m/z=579.3 (M+H).
2-(2-(iso-Butyl(2-methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethoxy)pyridin-3-yl)phenyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0608 (method A, 9%) as a light yellow solid. Rf=0.5 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 8.34 (d, J=2.5 Hz, 1H), 7.77 (dd, J=8.6, 2.5 Hz, 1H), 7.46 (dd, J=7.8, 1.7 Hz, 1H), 7.43 (d, J=1.7 Hz, 1H), 7.40 (d, J=7.8 Hz, 1H), 7.35 (s, 1H), 6.83 (d, J=8.6 Hz, 1H), 5.50 (s, 1H), 5.04 (brs, 4H), 4.49 (t, J=5.7 Hz, 2H), 3.80 (brs, 2H), 2.87 (t, J=5.7 Hz, 2H), 2.80-2.60 (m, 8H), 2.43 (s, 3H), 2.26 (s, 3H), 2.10-1.96 (m, 1H), 1.00 (d, J=6.6 Hz, 6H). 13C NMR (100 MHz, CDCl3) δ 171.42, 163.70, 163.30, 160.26, 151.03, 144.88, 144.16, 137.42, 137.33, 136.17, 132.96, 129.13, 127.42, 126.42, 111.36, 111.04, 82.90, 63.71, 59.87, 57.15, 55.06, 53.43, 46.02, 27.42, 20.67, 17.73; LCMS C30H39N9OS method (B) Rt=5.303 min, ESI+ m/z=574.3 (M+H).
N-((1-(3-((4-(4,6-Diaminopyrimidin-2-yl)thiazol-2-yl)(isobutyl)amino)-4-methylphenyl)-1H-1,2,3-triazol-4-yl)methyl) methanesulfonamide OR0609 (method A, 18%) as a light brown powder. Rf=0.5 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 8.51 (s, 1H), 7.88 (d, J=2.2 Hz, 1H), 7.82 (dd, J=8.3, 2.2 Hz, 1H), 7.60 (d, J=8.3 Hz, 1H), 7.30 (s, 1H), 5.53 (s, 1H), 4.45 (s, 2H), 3.91 (brs, 2H), 2.99 (s, 3H), 2.33 (s, 3H), 2.08-1.96 (m, 1H), 1.03 (d, J=6.7 Hz, 6H); 13C NMR (100 MHz, MeOD) δ 172.12, 165.62, 161.25, 152.19, 147.17, 145.86, 139.43, 137.65, 134.79, 122.65, 122.30, 121.25, 111.43, 83.02, 60.94, 40.62, 39.00, 28.74, 20.78, 17.85; LCMS C22H28N10O2S2 method (B) Rt=4.585 min, ESI+ m/z=529.2 (M+H).
2-(2-((2-Aminoethyl)(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0610 (method C, 74%), as a pale yellow solid. Rf=0.40 (DCM-MeOH—NH4OH, 80:18:2); 1H NMR (400 MHz, CDCl3) δ 7.49-7.47 (m, 2H), 7.43 (d, J=8.1 Hz, 2H), 7.31 (d, J=8.5 Hz, 1H), 7.25 (s, 1H), 7.20 (d, J=8.1 Hz, 2H), 5.71 (brs, 4H), 4.16 (brs, 1H), 3.80 (brs, 1H), 3.48-3.20 (m, 2H), 2.79-2.75 (m, 2H), 2.59-2.56 (m, 2H), 2.48 (brs, 8H), 2.33 (s, 3H), 2.14 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 170.65, 163.08, 162.63, 162.29, 143.80, 141.26, 139.88, 137.34, 134.63, 132.79, 129.41, 127.59, 126.98, 118.35, 115.44, 82.75, 60.07, 58.49, 54.87, 52.69, 45.72, 40.62, 33.08, 18.56, 17.41; LCMS C29H37N9S method (B) Rt=3.649 min, ESI+ m/z=544.2 (M+H).
tert-Butyl (2-((4-(4,6-diaminopyrimidin-2-yl)thiazol-2-yl)(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-bi-phenyl]-3-yl)amino)ethyl)carbamate OR0610-1 (method A, 31%) as a light brown solid. Rf=0.33 (DCM-MeOH—NH4OH, 90:9:1); LCMS C34H45N9O2S method (B) Rt=4.360 min, ESI+ m/z=644.4 (M+H).
2-(2-((3-Aminopropyl)(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0611 (method C, 50%) as a pale yellow solid. Rf=0.55 (DCM-MeOH—NH4OH, 80:18:2); 1H NMR (400 MHz, CDCl3) δ 7.47 (dd, J=8.1, 1.2 Hz, 1H), 7.41 (d, J=8.2 Hz, 2H), 7.31 (d, J=8.1 Hz, 1H), 7.30 (d, J=1.2 Hz, 1H), 7.21 (d, J=8.2 Hz, 2H), 7.15 (s, 1H), 5.51 (brs, 4H), 5.29 (s, 1H), 4.15 (brs, 1H), 3.81 (brs, 1H), 3.18-3.16 (m, 2H), 2.84-2.76 (m, 2H), 2.63-2.53 (m, 2H), 2.47 (brs, 8H), 2.28 (s, 3H), 2.13 (s, 3H), 2.01-1.97 (m, 2H); 13C NMR (100 MHz, CDCl3) δ 171.39, 164.09, 159.70, 150.42, 143.27, 141.36, 140.09, 137.45, 135.13, 132.80, 129.41, 127.47, 127.01, 110.34, 83.57, 60.45, 55.24, 53.56, 53.26, 50.17, 46.18, 33.31, 29.84, 17.44; LCMS C30H39N9S method (B) Rt=3.671 min, ESI+ m/z=558.3 (M+H).
tert-Butyl (3-((4-(4,6-diaminopyrimidin-2-yl)thiazol-2-yl)(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)propyl)carbamate OR0611-1 (method A, 39%) as a light brown solid. Rf=0.50 (DCM-MeOH—NH4OH, 90:9:1); LCMS C35H47N9O2S method (B) Rt=4.450 min, ESI+ m/z=658.4 (M+H).
2-[2-({2-Methyl-5-[6-(4-methyl-piperazin-1-ylcarbonyl)-pyridin-3-yl]-phenyl}-propyl-amino)-thiazol-4-yl]-pyrimidine-4,6-diamine OR0612 (method A, 2%) as a light yellow solid. Rf=0.5 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 8.78 (d, J=2.0 Hz, 1H), 7.96 (dd, J=8.1, 2.0 Hz, 1H), 7.72 (d, J=8.1 Hz, 1H), 7.54 (dd, J=7.9, 1.4 Hz, 1H), 7.47 (d, J=1.4 Hz, 1H), 7.45 (d, J=7.9 Hz, 1H), 7.35 (s, 1H), 5.53 (s, 1H), 5.26 (brs, 4H), 3.98 (brs, 2H), 3.87-3.83 (m, 2H), 3.70-3.66 (m, 2H), 2.55-2.51 (m, 2H), 2.45-2.41 (m, 2H), 2.33 (s, 3H), 2.27 (s, 3H), 1.66 (sext, J=7.3 Hz, 2H), 0.93 (t, J=7.3 Hz, 3H); LCMS C28H33N9OS method (B) Rt=4.071 min, ESI+ m/z=544.3 (M+H).
2-(2-((5-(2-Aminopyrimidin-5-yl)-2-methylphenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0613 (method B, 67%) as a light brown solid. Rf=0.45 (DCM-MeOH—NH4OH, 95:4.5:0.5); 1H NMR (400 MHz, MeOD) δ 8.54 (s, 2H), 7.54 (dd, J=8.0, 1.7 Hz, 1H), 7.50 (d, J=1.7 Hz, 1H), 7.47 (d, J=8.0 Hz, 1H), 7.29 (s, 1H), 5.54 (s, 1H), 3.98 (brs, 2H), 2.27 (s, 3H), 1.72 (sext, J=7.5 Hz, 2H), 0.97 (t, J=7.5 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 200.27, 193.20, 192.10, 188.30, 185.51, 179.17, 173.11, 165.67, 164.59, 162.29, 155.80, 155.25, 152.22, 139.76, 110.84, 83.11, 50.40, 45.71, 39.78; LCMS C21H23N9S method (B) Rt=4.620 min, ESI+ m/z=434.2 (M+H).
2-(2-((4-Methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0614 (method B, 35%) as a light yellow solid. Rf=0.22 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.74 (d, J=8.2 Hz, 2H), 7.67 (s, 1H), 7.62 (d, J=8.2 Hz, 2H), 7.40 (s, 1H), 7.23 (d, J=8.5 Hz, 1H), 7.19 (d, J=8.5 Hz, 1H), 5.39 (s, 1H), 5.13 (brs, 4H), 3.03-2.99 (m, 4H), 2.46-2.42 (m, 4H), 2.21 (s, 6H); 13C NMR (100 MHz, CDCl3) δ 166.59, 163.71, 159.74, 150.31, 145.04, 139.70, 138.20, 133.70, 131.96, 130.67, 128.45, 127.45, 123.37, 119.60, 110.58, 83.12, 67.13, 58.30, 54.04, 46.02, 45.71, 18.47, 17.66; LCMS C25H28N8O2S2 method (B) Rt=4.109 min, ESI+ m/z=537.2 (M+H).
3′-((4-(4,6-Diaminopyrimidin-2-yl)thiazol-2-yl)(propyl)amino)-4′-methyl-[1,1′-biphenyl]-4-sulfonamide OR0615 (method B, 36%) as a light brown solid. Rf=0.27 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 7.97 (d, J=8.4 Hz, 2H), 7.78 (d, J=8.4 Hz, 2H), 7.66 (dd, J=8.1, 1.8 Hz, 1H), 7.60 (d, J=1.8 Hz 1H), 7.50 (d, J=8.1 Hz, 1H), 7.29 (s, 1H), 5.55 (s, 1H), 3.98 (brs, 2H), 2.28 (s, 3H), 1.71 (sext, J=7.5 Hz, 2H), 0.97 (t, J=7.5 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 172.10, 165.00, 160.09, 150.99, 144.82, 144.72, 143.93, 140.59, 138.61, 134.05, 129.19, 128.50, 128.26, 127.88, 111.66, 82.69, 54.91, 22.25, 17.60, 11.63; LCMS C23H25N7O2S2 method (B) Rt=4.933 min, ESI+ m/z=496.2 (M+H).
N-(5-(3-((4-(4,6-Diaminopyrimidin-2-yl)thiazol-2-yl)(propyl)amino)-4-methylphenyl)pyridin-2-yl)-4-methylpiperazine-1-carboxamide OR0616 (method A, 19%) as a light yellow solid. Rf=0.5 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, DMSO-d6) δ 9.28 (brs, 1H), 8.59 (d, J=2.3 Hz, 1H), 8.03 (dd, J=8.8, 2.3 Hz, 1H), 7.86 (d, J=8.8 Hz, 1H), 7.68 (dd, J=7.9, 1.8 Hz, 1H), 7.67 (d, J=1.8 Hz, 1H), 7.49 (d, J=7.9 Hz, 1H), 7.17 (s, 1H), 6.03 (brs, 4H), 5.34 (s, 1H), 3.89 (brs, 2H), 3.50-3.42 (m, 4H), 2.33-2.26 (m, 4H), 2.21 (s, 3H), 2.18 (s, 3H), 1.62 (sext, J=7.4 Hz, 2H), 0.90 (t, J=7.4 Hz, 3H); LCMS C28H34N10OS method (B) Rt=4.237 min, ESI+ m/z=559.3 (M+H).
2-(2-((4-Methyl-4′-(morpholinosulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0617 (method B, 61%) as a light yellow solid. Rf=0.50 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 7.90 (d, J=8.5 Hz, 2H), 7.85 (d, J=8.5 Hz, 2H), 7.71 (dd, J=8.0, 1.7 Hz, 1H), 7.67 (d, J=1.7 Hz, 1H), 7.54 (d, J=8.0 Hz, 1H), 7.26 (s, 1H), 5.54 (s, 1H), 4.02 (brs, 2H), 3.73-3.69 (m, 4H), 3.02-2.98 (m, 4H), 2.31 (s, 3H), 1.63 (sext, J=7.3 Hz, 2H), 0.99 (t, J=7.3 Hz, 3H); 13C NMR (100 MHz, MeOD) S 172.00, 165.60, 161.21, 152.04, 145.94, 145.04, 140.32, 139.04, 135.37, 134.13, 129.78, 129.46, 128.61, 128.55, 111.03, 82.95, 67.23, 54.90, 47.46, 22.31, 17.62, 11.61; LCMS C27H31N7O3S2 method (B) Rt=5.538 min, ESI+ m/z=566.2 (M+H).
3′-((4-(4,6-Diaminopyrimidin-2-yl)thiazol-2-yl)(propyl)amino)-N,N,4′-trimethyl-[1,1′-biphenyl]-4-sulfonamide OR0618 (method B, 67%) as a light brown solid. Rf=0.55 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 7.88 (d, J=8.7 Hz, 2H), 7.85 (d, J=8.7 Hz, 2H), 7.71 (dd, J=7.9, 1.4 Hz, 1H), 7.66 (J=1.4 Hz, 1H), 7.53 (J=7.9 Hz, 1H), 7.25 (s, 1H), 5.54 (s, 1H), 4.02 (brs, 2H), 2.71 (s, 6H), 2.31 (s, 3H), 1.74 (sext, J=7.4 Hz, 2H), 0.99 (t, J=7.4 Hz, 3H). 13C NMR (100 MHz, MeOD) δ 172.00, 165.57, 161.16, 151.99, 145.63, 145.01, 140.38, 138.95, 135.44, 134.11, 132.26, 129.65, 129.41, 128.51, 111.05, 82.94, 54.88, 38.30, 22.30, 17.61, 11.61; LCMS C25H29N7O2S2 method (B) Rt=5.395 min, ESI+ m/z=524.2 (M+H).
2-(2-((4′-((Dimethylamino)methyl)-4-methyl-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0619 (method B, 61%) as a light yellow solid. Rf=0.50 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 7.63 (dd, J=8.0, 1.8 Hz, 1H), 7.60 (d, J=8.1 Hz, 2H), 7.54 (d, J=1.8 Hz, 1H), 7.47 (d, J=8.0 Hz, 1H), 7.40 (d, J=8.1 Hz, 2H), 7.23 (s, 1H), 5.54 (s, 1H), 4.01 (brs, 2H), 3.51 (s, 2H), 2.28 (s, 3H), 2.26 (s, 6H), 1.73 (sext, J=7.4 Hz, 2H), 0.99 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 172.18, 165.64, 161.34, 152.03, 144.75, 141.99, 140.42, 138.13, 137.33, 133.79, 131.38, 128.89, 128.18, 127.75, 110.91, 82.96, 64.53, 54.76, 45.22, 22.30, 17.50, 11.61; LCMS C26H31N7S method (B) Rt=4.385 min, ESI+ m/z=474.3 (M+H).
2-(2-((2-Methyl-5-(pyridin-3-yl)phenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0620 (method B, 86%) as a light brown solid. Rf=0.50 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 8.81 (d, J=1.8 Hz, 1H), 8.52 (dd, J=4.9, 1.8 Hz, 1H), 8.10 (dt, J=8.0, 1.8 Hz, 1H), 7.67 (dd, J=7.8, 1.9 Hz, 1H), 7.63 (d, J=1.9 Hz, 1H), 7.55 (d, J=7.8 Hz, 1H), 7.52 (dd, J=8.0, 4.9 Hz, 1H), 7.25 (s, 1H), 5.54 (s, 1H), 4.01 (brs, 2H), 2.31 (s, 3H), 1.74 (sext, J=7.4 Hz, 2H), 0.99 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 171.98, 165.61, 161.25, 152.05, 149.00, 148.19, 145.12, 138.79, 138.49, 137.44, 136.37, 134.21, 129.17, 128.33, 125.57, 111.01, 82.96, 54.92, 22.30, 17.61, 11.60; LCMS C22H23N7S method (B) Rt=4.544 min, ESI+ m/z=418.2 (M+H).
(5-(3-((4-(4,6-Diaminopyrimidin-2-yl)thiazol-2-yl)(propyl)amino)-4-methylphenyl)pyridin-2-yl)(morpholino) methanone OR0621 (method B, 71%) as a light yellow solid. Rf=0.50 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 8.88 (d, J=2.0 Hz, 1H), 8.21 (dd, J=8.2, 2.0 Hz, 1H), 7.73 (dd, J=8.0, 1.8 Hz, 1H), 7.72 (d, J=8.2 Hz, 1H), 7.69 (d, J=1.8 Hz, 1H), 7.56 (d, J=8.0 Hz, 1H), 7.25 (s, 1H), 5.53 (s, 1H), 4.02 (brs, 2H), 3.79 (brs, 4H), 3.70-3.64 (m, 2H), 3.62-3.56 (m, 2H), 2.32 (s, 3H), 1.74 (sext, J=7.4 Hz, 2H), 0.99 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 171.91, 169.24, 165.61, 161.24, 153.30, 152.08, 147.72, 145.20, 139.29, 138.03, 137.87, 136.88, 134.30, 129.36, 128.41, 124.96, 111.02, 82.97, 67.93, 54.94, 44.00, 22.30, 17.65, 11.60; LCMS C27H30N8O2S method (B) Rt=4.835 min, ESI+ m/z=531.2 (M+H).
Methyl 5-(3-((4-(4,6-diaminopyrimidin-2-yl)thiazol-2-yl)(propyl)amino)-4-methylphenyl)picolinate OR0622 (method B, 42%) as a light brown solid. Rf=0.50 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 8.95 (dd, J=2.0, 0.6 Hz, 1H), 8.27 (dd, J=8.2, 2.0 Hz, 1H), 8.22 (dd, J=8.2, 0.6 Hz, 1H), 7.75 (dd, J=8.0, 1.8 Hz, 1H), 7.73 (d, J=1.8 Hz, 1H), 7.58 (d, J=8.0 Hz, 1H), 7.26 (s, 1H), 5.54 (s, 1H), 4.02 (brs, 2H), 4.00 (s, 3H) 2.33 (s, 3H), 1.74 (sext, J=7.4 Hz, 2H), 0.99 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 171.88, 166.44, 165.59, 161.19, 152.06, 148.65, 147.36, 145.28, 140.31, 139.80, 137.46, 136.88, 134.38, 129.52, 128.54, 126.61, 111.07, 82.96, 54.99, 53.24, 22.30, 17.69, 11.60; LCMS C24H25N7O2S method (B) Rt=5.062 min, ESI+ m/z=476.3 (M+H).
N-(5-(3-((4-(4,6-Diaminopyrimidin-2-yl)thiazol-2-yl)(propyl)amino)-4-methylphenyl)pyridin-2-yl)-4-methylpiperazine-1-sulfonamide OR0625 (method A, 18%) as a white powder. Rf=0.5 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, DMSO-d6) 1H NMR (400 MHz, DMSO-d6) δ 8.51 (d, J=2.0 Hz, 1H), 8.12 (dd, J=8.8, 2.0 Hz, 1H), 7.71 (dd, J=8.4, 1.8 Hz, 1H), 7.70 (d, J=1.8 Hz, 1H), 7.52 (d, J=8.8 Hz, 1H), 7.41 (s, 1H), 7.21 (d, J=8.4 Hz, 1H), 7.03 (s, 4H), 5.48 (s, 1H), 3.97 (brs, 2H), 3.25-3.21 (m, 4H), 2.56-2.52 (m, 4H), 2.28 (s, 3H), 2.22 (s, 3H), 1.62 (sext, J=7.4 Hz, 2H), 0.93 (t, J=7.4 Hz, 3H); LCMS C27H34N10O2S2 method (B) Rt=4.182 min, ESI+ m/z=595.3 (M+H). 2-(2-((5-(1H-Indol-5-yl)-2-methylphenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0626 (method B, 82%) as a light yellow solid. Rf=0.50 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 7.78 (d, J=1.8 Hz, 1H), 7.63 (dd, J=7.9, 1.8 Hz, 1H), 7.53 (d, J=1.6 Hz, 1H), 7.44 (d, J=7.9 Hz, 1H), 7.43 (d, J=8.2 Hz, 1H), 7.36 (dd, J=8.2, 1.6 Hz, 1H), 7.25 (s, 1H), 7.24 (d, J=2.8 Hz, 1H), 6.50 (d, J=2.8 Hz, 1H), 5.54 (s, 1H), 4.00 (brs, 2H), 2.27 (s, 3H), 1.75 (sext, J=7.4 Hz, 2H), 0.99 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 172.44, 165.37, 160.86, 151.48, 144.48, 144.22, 137.37, 135.56, 133.49, 132.36, 130.07, 128.85, 128.40, 126.44, 121.60, 119.39, 112.58, 111.16, 102.85, 82.81, 58.32, 54.72, 22.30, 18.36, 17.40, 11.64; LCMS C25H25N7S method (B) Rt=5.452 min, ESI+ m/z=456.2 (M+H).
2-(2-((2-Methyl-5-(6-((4-methylpiperazin-1-yl)sulfonyl)pyridin-3-yl)phenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0627 (method A, 22% or method B, 71%), as a light yellow powder. Rf=0.50 (DCM-MeOH—NH4OH, 90:10:1); 1H NMR (400 MHz, MeOD) δ 9.00 (d, J=1.8 Hz, 1H), 8.31 (dd, J=8.2, 1.8 Hz, 1H), 8.01 (d, J=8.2 Hz, 1H), 7.76 (dd, J=7.8, 1.9 Hz, 1H), 7.74 (d, J=1.9 Hz, 1H), 7.58 (d, J=7.8 Hz, 1H), 7.26 (s, 1H), 5.53 (s, 1H), 4.02 (brs, 2H), 3.35-3.30 (m, 4H), 2.54-2.46 (m, 4H), 2.33 (s, 3H), 2.28 (s, 3H), 1.74 (sext, J=7.4 Hz, 2H), 0.99 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 171.84, 165.64, 161.29, 155.57, 152.17, 149.29, 145.31, 140.03, 139.92, 137.40, 137.14, 134.41, 129.64, 128.60, 124.55, 111.02, 82.99, 55.38, 55.00, 47.30, 45.87, 22.30, 17.70, 11.60; LCMS C27H33N9O2S2 method (B) Rt=4.346 min, ESI+ m/z=580.3 (M+H).
2-(2-((2-Methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethoxy)pyridin-3-yl)phenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0629 (method A, 31%) as a white powder. Rf=0.5 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 8.34 (d, J=2.3 Hz, 1H), 7.75 (dd, J=8.6, 2.3 Hz, 1H), 7.45 (dd, J=7.9, 1.8 Hz, 1H), 7.39 (d, J=7.9 Hz, 1H), 7.38 (d, J=1.8 Hz, 1H), 7.31 (s, 1H), 6.81 (d, J=8.6 Hz, 1H), 5.42 (s, 1H), 4.89 (brs, 4H), 4.47 (t, J=5.8 Hz, 2H), 3.98 (brs, 2H), 2.82 (t, J=5.8 Hz, 2H), 2.64 (brs, 4H), 2.51 (brs, 4H), 2.30 (s, 3H), 2.25 (s, 3H), 1.65 (sext, J=7.4 Hz, 2H), 0.91 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.95, 163.76, 163.31, 160.39, 151.04, 144.88, 143.52, 137.64, 137.33, 136.53, 132.76, 129.09, 127.63, 126.59, 111.33, 110.87, 82.92, 63.71, 57.15, 55.05, 53.72, 53.40, 46.01, 21.32, 17.50, 11.39; LCMS C29H37N9OS method (B) Rt=4.379 min, ESI+m/z=560.3 (M+H).
2-((5-(3-((4-(4,6-Diaminopyrimidin-2-yl)thiazol-2-yl)(propyl)amino)-4-methylphenyl)pyridin-2-yl)oxy)-1-(4-methylpiperazin-1-yl)ethan-1-one OR0630 (method A, 3%) as a white powder. Rf=0.5 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 8.30 (d, J=2.4 Hz, 1H), 7.80 (dd, J=8.6, 2.4 Hz, 1H), 7.45 (dd, J=7.9, 1.7 Hz, 1H), 7.38 (d, J=7.9 Hz, 1H), 7.37 (d, J=1.7 Hz, 1H), 7.36 (s, 1H), 6.96 (d, J=8.6 Hz, 1H), 5.56 (s, 1H), 5.47 (brs, 4H), 5.05 (s, 2H), 3.97 (brs, 2H), 3.70-3.62 (m, 2H), 3.58-3.50 (m, 2H), 2.51-2.43 (m, 2H), 2.45-2.37 (m, 2H), 2.32 (s, 3H), 2.24 (s, 3H), 1.65 (sext, J=7.4 Hz, 2H), 0.93 (t, J=7.4 Hz, 3H); LCMS C29H35N9O2S method (B) Rt=4.361 min, ESI+ m/z=574.3 (M+H).
2-(2-(Methyl(4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0631 (method B, 67%) as a light yellow solid. Rf=0.47 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 7.89 (d, J=8.6 Hz, 2H), 7.84 (d, J=8.6 Hz, 2H), 7.71 (d, J=1.9 Hz, 1H), 7.69 (dd, J=7.7, 1.9 Hz, 1H), 7.53 (d, J=7.7 Hz, 1H), 7.29 (s, 1H), 5.54 (s, 1H), 3.60 (s, 3H), 3.09-3.01 (m, 4H), 2.54-2.46 (m, 4H), 2.32 (s, 3H), 2.25 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 171.88, 165.64, 161.18, 152.13, 146.67, 145.82, 140.49, 138.39, 135.59, 134.02, 129.67, 128.56, 128.49, 128.31, 111.39, 82.98, 55.08, 46.95, 45.75, 40.15, 17.37; LCMS C26H30N8O2S2 method (B) Rt=4.294 min, ESI+ m/z=551.2 (M+H).
2-(2-(Ethyl(4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidine-4,6-di-amine OR0632 (method B, 58%) as a light brown solid. Rf=0.36 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 7.89 (d, J=8.6 Hz, 2H), 7.84 (d, J=8.6 Hz, 2H), 7.71 (dd, J=8.0, 1.8 Hz, 1H), 7.66 (d, J=1.8 Hz, 1H), 7.54 (d, J=8.0 Hz, 1H), 7.26 (s, 1H), 5.54 (s, 1H), 4.13 (brs, 2H), 3.09-3.01 (m, 4H), 2.54-2.46 (m, 4H), 2.31 (s, 3H), 2.25 (s, 3H), 1.28 (t, J=7.2 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 171.66, 165.65, 161.30, 152.12, 145.84, 144.65, 140.31, 139.18, 135.58, 134.06, 129.70, 129.56, 128.58, 128.55, 110.99, 82.98, 55.08, 47.75, 46.95, 45.75, 17.58, 13.55, LCMS C27H32N8O2S2 method (B) Rt=4.418 min, ESI+ m/z=565.2 (M+H).
2-(2-((5-Fluoro-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0633 (method A, 11%) as a light brown solid. Rf=0.5 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.81 (d, J=8.4 Hz, 2H), 7.68 (d, J=8.4 Hz, 2H), 7.40 (s, 1H), 7.32 (dd, J=10.2, 1.6 Hz, 1H), 7.31 (d, J=1.6 Hz, 1H), 5.57 (s, 1H), 5.40 (brs, 4H), 4.01 (brs, 2H), 3.10-3.02 (m, 4H), 2.53-2.45 (m, 4H), 2.26 (s, 3H), 2.19 (s, 3H), 1.66 (sext, J=7.4 Hz, 2H), 0.95 (t, J=7.4 Hz, 3H); 19F NMR (376 MHz, CDCl3) δ −110.83; LCMS C2SH33FN8O2S2 method (B) Rt=4.384 min, ESI+ m/z=597.2 (M+H).
2-(2-((3′-Methoxy-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0635 (method A, 36%) as light brown powder. Rf=0.5 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.91 (d, J=8.1 Hz, 1H), 7.53 (dd, J=8.1, 1.9 Hz, 1H), 7.45 (d, J=1.9 Hz, 1H), 7.43 (d, J=8.2 Hz, 1H), 7.35 (s, 1H), 7.20 (dd, J=8.2, 1.4 Hz, 1H), 7.12 (d, J=1.4 Hz, 1H), 5.49 (s, 1H), 5.10 (brs, 4H), 3.99 (brs, 2H), 3.96 (s, 3H), 3.31-3.22 (m, 4H), 2.50-2.42 (m, 4H), 2.29 (s, 3H), 2.28 (s, 3H), 1.66 (sext, J=7.4 Hz, 2H), 0.94 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.91, 163.31, 159.37, 157.46, 150.25, 146.51, 143.59, 139.34, 138.09, 132.85, 132.48, 128.36, 127.30, 125.02, 119.05, 111.33, 110.73, 82.80, 56.20, 54.84, 53.94, 46.02, 45.98, 21.34, 17.64, 11.42; LCMS C29H36FN8O3S2 method (B) Rt=4.258 min, ESI+ m/z=609.3 (M+H).
2-(2-(iso-Propyl(4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0636 (method B, 29%) as a light brown solid. Rf=0.41 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 7.88 (d, J=8.0 Hz, 2H), 7.84 (d, J=8.0 Hz, 2H), 7.73 (dd, J=8.0, 1.9 Hz, 1H), 7.57 (d, J=1.9 Hz, 1H), 7.55 (d, J=8.0 Hz, 1H), 7.24 (s, 1H), 5.54 (s, 1H), 5.21 (sept, J=6.7 Hz, 1H), 3.09-3.01 (m, 4H), 2.54-2.46 (m, 4H), 2.32 (s, 3H), 2.25 (s, 3H), 1.29-1.28 (m, 6H); 13C NMR (100 MHz, MeOD) δ 171.77, 165.62, 161.37, 152.12, 145.82, 142.12, 140.70, 140.05, 135.57, 134.11, 130.95, 129.73, 128.86, 128.57, 110.77, 82.97, 55.07, 52.86, 46.94, 45.75, 21.46, 18.24; LCMS C28H34FN8O2S2 method (B) Rt=4.311 min, ESI+ m/z=579.3 (M+H).
2-(2-(iso-Butyl(4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0637 (method B, 70%) as a light brown solid. Rf=0.45 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 7.88 (d, J=8.8 Hz, 2H), 7.85 (d, J=8.8 Hz, 2H), 7.69 (dd, J=8.0, 1.7 Hz, 1H), 7.67 (d, J=1.7 Hz, 1H), 7.53 (d, J=8.0 Hz, 1H), 7.26 (s, 1H), 5.53 (s, 1H), 3.92 (brs, 2H), 3.09-3.01 (m, 4H), 2.54-2.46 (m, 4H), 2.31 (s, 3H), 2.25 (s, 3H), 2.06-1.96 (m, 1H), 1.03 (d, J=6.6 Hz, 6H); 13C NMR (100 MHz, MeOD) δ 172.66, 165.63, 161.33, 152.13, 145.90, 145.49, 140.15, 138.81, 135.57, 134.36, 129.73, 129.26, 128.58, 128.41, 111.14, 83.00, 60.51, 55.08, 46.95, 45.75, 28.72, 20.82, 17.80; LCMS C29H36FN8O2S2 method (B) Rt=4.369 min, ESI+ m/z=593.3 (M+H).
2-(2-((4′-((4-Ethylpiperazin-1-yl)sulfonyl)-4-methyl-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0638 (method A, 36%) as a white solid. Rf=0.5 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.80 (d, J=8.5 Hz, 2H), 7.69 (d, J=8.5 Hz, 2H), 7.53 (dd, J=7.9, 1.9 Hz, 1H), 7.45 (d, J=1.9 Hz, 1H), 7.44 (d, J=7.9 Hz, 1H), 7.35 (s, 1H), 5.49 (s, 1H), 5.09 (brs, 4H), 4.00 (brs, 2H), 3.10-3.02 (m, 4H), 2.57-2.49 (m, 4H), 2.39 (q, J=7.2 Hz, 2H), 2.28 (s, 3H), 1.67 (sext, J=7.4 Hz, 2H), 1.02 (t, J=7.2 Hz, 3H), 0.94 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.91, 163.32, 159.40, 150.31, 144.63, 143.58, 139.05, 138.01, 133.88, 132.96, 128.63, 128.55, 127.56, 127.30, 111.30, 82.81, 53.83, 52.02, 51.91, 46.23, 21.35, 17.61, 12.03, 11.42; LCMS C29H36FN8O2S2 method (B) Rt=4.316 min, ESI+ m/z=593.3 (M+H).
2-(2-((4-Methyl-4′-(piperazin-1-ylsulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0639 (method C, 53%) as a white solid. Rf=0.18 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.83 (d, J=8.2 Hz, 2H), 7.74 (d, J=8.2 Hz, 2H), 7.57 (dd, J=7.9, 1.9 Hz, 1H), 7.51 (d, J=1.9 Hz, 1H), 7.47 (d, J=7.9 Hz, 1H), 7.38 (s, 1H), 5.50 (s, 1H), 5.00 (brs, 4H), 4.01 (brs, 2H), 3.10-3.02 (m, 4H), 3.01-2.94 (m, 4H), 2.30 (s, 3H), 1.69 (sext, J=7.2 Hz, 2H), 0.96 (t, J=7.2 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 179.89, 170.95, 163.53, 159.84, 150.64, 144.57, 143.59, 139.00, 138.05, 134.28, 132.93, 128.59, 128.52, 127.57, 127.28, 111.18, 82.92, 53.89, 46.84, 45.32, 35.43, 21.34, 17.63, 11.43; LCMS C27H32N8O2S2 method (B) Rt=4.272 min, ESI+ m/z=565.3 (M+H).
tert-Butyl 4-((3′-((4-(4,6-diaminopyrimidin-2-yl)thiazol-2-yl)(propyl)amino)-4′-methyl-[1,1′-biphenyl]-4-yl)sulfonyl) piperazine-1-carboxylate OR0639-1 (method A, 30%) as a light brown solid. 1H NMR (400 MHz, CDCl3) δ 7.80 (d, J=8.5 Hz, 2H), 7.72 (d, J=8.5 Hz, 2H), 7.56 (dd, J=7.9, 1.9 Hz, 1H), 7.48 (d, J=1.9 Hz, 1H), 7.46 (d, J=7.9 Hz, 1H), 7.40 (s, 1H), 5.76 (brs, 4H), 5.66 (s, 1H), 4.04 (brs, 2H), 3.55-3.47 (m, 4H), 3.05-2.97 (m, 4H), 2.29 (s, 3H), 1.69 (sext, J=7.4 Hz, 2H), 1.39 (s, 9H), 0.97 (t, J=7.4 Hz, 3H); LCMS C32H40N8O4S2 method (B) Rt=5.632 min, ESI+ m/z=665.3 (M+H).
2-(2-((4-Methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)oxazol-4-yl)pyrimidine-4,6-diamine OR0640 (method A, 34%) as a light brown solid. Rf=0.50 (DCM-MeOH—NH4OH, 90:10:1); 1H NMR (400 MHz, MeOD) δ 7.86 (d, J=8.4 Hz, 2H), 7.83 (s, 1H), 7.82 (d, J=8.4 Hz, 2H), 7.61 (d, J=1.6 Hz, 1H), 7.59 (dd, J=8.4, 1.6 Hz, 1H), 7.45 (d, J=8.4 Hz, 1H), 5.48 (s, 1H), 3.89-3.81 (m, 2H), 3.08-3.00 (m, 4H), 2.53-2.45 (m, 4H), 2.25 (s, 3H), 2.24 (s, 3H), 1.74 (sext, J=7.4 Hz, 2H), 0.97 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 165.48, 162.13, 159.85, 146.19, 142.81, 141.16, 139.57, 138.48, 135.45, 134.97, 133.31, 129.64, 128.61, 128.57, 127.55, 82.92, 55.09, 54.80, 46.95, 45.75, 22.39, 17.74, 11.51; LCMS C28H34N8O3S method (B) Rt=4.676 min, ESI+ m/z=564.3 (M+H).
2-(2-((2′-Chloro-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0641 (method A, 28%) as a light yellow solid. Rf=0.50 (DCM-MeOH—NH4OH, 90:10:1); 1H NMR (400 MHz, MeOD) δ 7.90 (d, J=1.7 Hz, 1H), 7.77 (dd, J=8.1, 1.7 Hz, 1H), 7.67 (d, J=8.1 Hz, 1H), 7.54 (d, J=7.6 Hz, 1H), 7.46 (dd, J=7.6, 1.8 Hz, 1H), 7.44 (d, J=1.8 Hz, 1H), 7.26 (s, 1H), 5.53 (s, 1H), 3.99 (brs, 2H), 3.13-3.05 (m, 4H), 2.56-2.48 (m, 4H), 2.32 (s, 3H), 2.27 (s, 3H), 1.73 (sext, J=7.4 Hz, 2H), 0.98 (t, J=7.4 Hz, 3H). 13C NMR (100 MHz, MeOD) δ 171.97, 165.58, 161.20, 152.02, 145.23, 144.10, 139.09, 138.78, 137.57, 134.54, 133.48, 133.36, 131.77, 130.59, 130.21, 127.71, 111.10, 82.96, 55.07, 54.72, 46.94, 45.76, 22.26, 17.64, 11.61; LCMS C28H33ClN8O2S2 method (B) Rt=4.434 min, ESI+ m/z=613.2 (M+H).
2-(2-((4-Methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-2′-(trifluoromethyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0642 (method A, 31% or method B, 56%) as a light yellow powder. Rf=0.27 (DCM-MeOH—NH4OH, 95:5:0.5); 1H NMR (400 MHz, MeOD) δ 8.11 (d, J=1.5 Hz, 1H), 8.07 (dd, J=8.0, 1.5 Hz, 1H), 7.70 (d, J=8.0 Hz, 1H), 7.52 (d, J=7.9 Hz, 1H), 7.35 (dd, J=7.9, 1.4 Hz, 1H), 7.31 (d, J=1.4 Hz, 1H), 7.26 (s, 1H), 5.53 (s, 1H), 4.00 (brs, 2H), 3.14-3.06 (m, 4H), 2.57-2.49 (m, 4H), 2.33 (s, 3H), 2.27 (s, 3H), 1.70 (sext, J=7.4 Hz, 2H), 0.98 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 171.96, 165.63, 161.29, 152.08, 146.18, 143.87, 139.13, 137.11, 134.77, 133.23, 132.28, 131.14, 130.53, 130.22, 130.12, 126.56, 111.07, 82.98, 55.05, 54.64, 46.90, 45.75, 22.20, 17.60, 11.58; LCMS C29H33F3N8O2S2 method (B) Rt=4.470 min, ESI+ m/z=647.2 (M+H).
2-(2-((2′-Fluoro-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0643 (method A, 32%) as a light brown powder. Rf=0.55 (DCM-MeOH—NH4OH, 90:10:1); 1H NMR (400 MHz, MeOD) δ 7.81-7.75 (m, 1H), 7.68-7.52 (m, 5H), 7.26 (s, 1H), 5.53 (s, 1H), 4.00 (brs, 2H), 3.13-3.05 (m, 4H), 2.56-2.48 (m, 4H), 2.32 (s, 3H), 2.27 (s, 3H), 1.73 (sext, J=7.4 Hz, 2H), 0.99 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 171.92, 165.63, 161.29, 159.37, 152.12, 144.59, 139.43, 137.77, 135.04, 133.97, 133.78, 132.79, 131.41, 130.35, 125.29, 117.08, 111.03, 82.98, 55.08, 54.81, 46.94, 45.76, 22.26, 17.67, 11.59; LCMS C28H33FN8O2S2 method (B) Rt=4.322 min, ESI+ m/z=597.3 (M+H).
2-(2-((2′-Fluoro-6′-methoxy-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0644 (method A, 24%) as a light yellow solid. Rf=0.55 (DCM-MeOH—NH4OH, 90:10:1); 1H NMR (400 MHz, MeOD) δ 7.49 (d, J=7.9 Hz, 1H), 7.39 (d, J=7.9 Hz, 1H), 7.35 (s, 1H), 7.28-7.20 (m, 3H), 5.53 (s, 1H), 3.99 (brs, 2H), 3.88 (s, 3H), 3.15-3.07 (m, 4H), 2.57-2.49 (m, 4H), 2.31 (s, 3H), 2.28 (s, 3H), 1.72 (sext, J=7.4 Hz, 2H), 0.98 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 172.13, 165.61, 162.39, 161.30, 159.88, 152.01, 143.84, 138.53, 137.89, 133.05, 132.91, 131.87, 130.92, 123.26, 111.03, 108.99, 107.48, 82.96, 57.18, 55.11, 54.57, 46.98, 45.77, 22.18, 17.62, 11.61; LCMS C29H35FN8O3S2 method (B) Rt=4.388 min, ESI+ m/z=627.3 (M+H).
2-(2-((2′-Methoxy-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0645 (method A, 28%) as a light brown solid. Rf=0.68 (DCM-MeOH—NH4OH, 90:10:1); 1H NMR (400 MHz, MeOD) δ 7.57 (d, J=7.9 Hz, 1H), 7.52 (dd, J=7.9, 1.7 Hz, 1H), 7.51 (d, J=1.7 Hz, 1H), 7.47 (d, J=7.9 Hz, 1H), 7.44 (dd, J=7.9, 1.7 Hz, 1H), 7.37 (d, J=1.7 Hz, 1H), 7.25 (s, 1H), 5.54 (s, 1H), 4.02 (brs, 2H), 3.89 (s, 3H), 3.12-3.04 (m, 4H), 2.56-2.50 (m, 4H), 2.29 (s, 3H), 2.26 (s, 3H), 1.73 (sext, J=7.6 Hz, 2H), 0.99 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 172.16, 165.50, 161.08, 158.24, 151.83, 143.86, 137.99, 137.83, 136.89, 135.50, 133.12, 132.12, 131.97, 130.75, 121.54, 111.54, 111.08, 82.90, 56.59, 55.11, 54.63, 46.98, 45.76, 22.21, 17.57, 11.63; LCMS C29H36FN8O3S2 method (B) Rt=4.359 min, ESI+ m/z=609.3 (M+H).
2-(2-((4-Methyl-4′-(piperidin-4-ylsulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0646 (method A, 28%) as a light yellow solid. Rf=0.20 (DCM-MeOH—NH4OH, 90:10:1); 1H NMR (400 MHz, MeOD) δ 7.95 (d, J=8.6 Hz, 2H), 7.91 (d, J=8.6 Hz, 2H), 7.71 (dd, J=8.0, 1.9 Hz, 1H), 7.66 (d, J=1.9 Hz, 1H), 7.55 (d, J=8.0 Hz, 1H), 7.25 (s, 1H), 5.54 (s, 1H), 4.01 (brs, 2H), 3.32-3.24 (m, 1H), 3.12-3.04 (m, 2H), 2.59-2.51 (m, 2H), 2.31 (s, 3H), 1.99-1.91 (m, 2H), 1.72 (sext, J=7.4 Hz, 2H), 1.64-1.53 (m, 2H), 0.99 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 171.95, 165.63, 161.27, 152.11, 146.75, 145.05, 140.20, 139.23, 136.81, 134.17, 130.94, 129.54, 128.66, 128.59, 111.00, 82.99, 62.42, 54.89, 45.52, 26.62, 22.30, 17.63, 11.61; LCMS C28H33N7O2S2 method (B) Rt=4.266 min, ESI+ m/z=564.2 (M+H).
2-(2-((2′,6′-Difluoro-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0647 (method A, 27%) as a light yellow solid. Rf=0.70 (DCM-MeOH—NH4OH, 90:10:1); 1H NMR (400 MHz, MeOD) δ 7.57 (d, J=8.0 Hz, 1H), 7.54-7.48 (m, 3H), 7.48 (s, 1H), 7.26 (s, 1H), 5.53 (s, 1H), 3.99 (brs, 2H), 3.16-3.08 (m, 4H), 2.57-2.49 (m, 4H), 2.33 (s, 3H), 2.28 (s, 3H), 1.72 (sext, J=7.4 Hz, 2H), 0.98 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 171.90, 165.62, 162.49, 161.27, 159.90, 152.09, 144.38, 139.94, 138.60, 133.52, 132.70, 131.55, 128.50, 112.64, 111.10, 82.98, 55.07, 54.73, 46.92, 45.77, 22.21, 17.72, 11.58; LCMS C28H32N8O2S2 method (B) Rt=4.396 min, ESI+ m/z=615.2 (M+H).
2-(2-((4′-((4-Aminopiperidin-1-yl)sulfonyl)-4-methyl-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0648 (method C, 90%) as a light yellow solid. Rf=0.18 (DCM-MeOH—NH4OH, 90:9:1). 1H NMR (400 MHz, MeOD) δ 7.84 (d, J=8.6 Hz, 2H), 7.81 (d, J=8.6 Hz, 2H), 7.67 (dd, J=8.0, 1.5 Hz, 1H), 7.62 (d, J=1.5 Hz, 1H), 7.51 (d, J=8.0 Hz, 1H), 7.24 (s, 1H), 5.54 (s, 1H), 3.99 (brs, 2H), 3.74-3.66 (m, 2H), 2.60-2.52 (m, 1H), 2.43-2.35 (m, 2H), 2.28 (s, 3H), 1.89-1.81 (m, 2H), 1.71 (sext, J=7.4 Hz, 2H), 1.46-1.34 (m, 2H), 0.96 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 171.92, 165.62, 161.29, 152.14, 145.55, 144.99, 140.27, 138.95, 136.32, 134.13, 129.50, 129.38, 128.54, 128.48, 111.01, 83.03, 54.86, 48.72, 46.43, 34.80, 22.29, 17.64, 11.63; LCMS C28H34N8O2S2 method (B) Rt=4.317 min, ESI+ m/z=579.3 (M+H).
tert-Butyl (1-((3′-((4-(4,6-diaminopyrimidin-2-yl)thiazol-2-yl)(propyl)amino)-4′-methyl-[1,1′-biphenyl]-4-yl)sulfonyl) piperidin-4-yl)carbamate OR0648-1 (method A, 20%) as a light brown solid. Rf=0.20 (DCM-MeOH—NH4OH, 95:5:0.5); 1H NMR (400 MHz, CDCl3) δ 7.80 (d, J=8.5 Hz, 2H), 7.72 (d, J=8.5 Hz, 2H), 7.57 (dd, J=7.9, 1.8 Hz, 1H), 7.50 (d, J=1.8 Hz, 1H), 7.46 (d, J=7.9 Hz, 1H), 7.41 (s, 1H), 5.70 (brs, 4H), 5.65 (s, 1H), 4.42 (brs, 1H), 4.05 (brs, 2H), 3.73 (brs, 1H), 3.40 (brs, 1H), 2.53-2.45 (m, 2H), 2.29 (s, 3H), 2.02-1.96 (m, 2H), 1.71 (sext, J=7.4 Hz, 2H), 1.55-1.47 (m, 2H), 1.40 (s, 9H), 0.98 (t, J=7.4 Hz, 3H); LCMS C33H42N8O4S2 method (B) Rt=5.495 min, ESI+ m/z=679.3 (M+H).
2-(4-((3′-((4-(4,6-Diaminopyrimidin-2-yl)thiazol-2-yl)(propyl)amino)-4′-methyl-[1,1′-biphenyl]-4-yl)sulfonyl) piperazin-1-yl)acetic acid OR0649 (method A, 20%) as a white solid. Rf=0.20 (DCM-MeOH—NH4OH, 80:18:2); 1H NMR (400 MHz, MeOD) δ 7.89 (d, J=8.7 Hz, 2H), 7.85 (d, J=8.7 Hz, 2H), 7.74 (dd, J=8.0, 1.9 Hz, 1H), 7.66 (d, J=1.9 Hz, 1H), 7.57 (s, 1H), 7.56 (d, J=8.0 Hz, 1H), 5.61 (s, 1H), 4.01 (brs, 2H), 3.17 (s, 2H), 3.16-3.12 (m, 4H), 2.86-2.80 (m, 4H), 2.31 (s, 3H), 1.74 (sext, J=7.4 Hz, 2H), 1.01 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 174.37, 172.57, 162.36, 154.84, 146.30, 145.71, 144.52, 140.49, 138.81, 135.64, 134.31, 129.76, 129.26, 128.85, 128.64, 114.62, 81.17, 61.32, 54.98, 53.18, 46.41, 22.21, 17.56, 11.73; LCMS C29H34N8O4S2 method (B) Rt=4.505 min, ESI+ m/z=623.3 (M+H).
2-(2-((4-Methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-3′-(trifluoromethoxy)-[1,1′-biphenyl]-3-yl)(propyl)amino) thiazol-4-yl)pyrimidine-4,6-diamine OR0650 (method A, 40%) as a light brown solid. Rf=0.20 (DCM-MeOH—NH4OH, 95:5:0.5); 1H NMR (400 MHz, MeOD) δ 8.04 (d, J=8.3 Hz, 1H), 7.81 (dd, J=8.3, 1.7 Hz, 1H), 7.73-7.72 (m, 1H), 7.69 (dd, J=7.9, 2.0 Hz, 1H), 7.66 (d, J=2.0 Hz, 1H), 7.56 (d, J=7.9 Hz, 1H), 7.26 (s, 1H), 5.54 (s, 1H), 4.01 (brs, 2H), 3.27-3.19 (m, 4H), 2.53-2.45 (m, 4H), 2.31 (s, 3H), 2.28 (s, 3H), 1.73 (sext, J=7.4 Hz, 2H), 0.99 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 171.84, 165.58, 161.18, 152.07, 148.09, 147.72, 145.17, 139.95, 138.92, 134.35, 133.68, 130.03, 129.48, 128.48, 126.65, 122.98, 120.66, 111.09, 82.97, 55.37, 54.97, 46.59, 45.86, 22.29, 17.70, 11.60; LCMS C29H33F3N8O3S2 method (B) Rt=4.530 min, ESI+ m/z=663.2 (M+H).
N-(2-Aminoethyl)-3′-((4-(4,6-diaminopyrimidin-2-yl)thiazol-2-yl)(propyl)amino)-4′-methyl-[1,1′-biphenyl]-4-sulfonamide OR0651 (method C, 100%) as a light yellow solid. Rf=0.22 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 7.97 (d, J=8.5 Hz, 2H), 7.87 (d, J=8.5 Hz, 2H), 7.75 (dd, J=8.0, 1.7 Hz, 1H), 7.69 (s, 1H), 7.66 (d, J=1.7 Hz, 1H), 7.58 (d, J=8.0 Hz, 1H), 5.65 (s, 1H), 4.22 (brs, 1H), 3.84 (brs, 1H), 3.18-3.12 (m, 2H), 3.12-3.04 (m, 2H), 2.32 (s, 3H), 1.75 (sext, J=7.4 Hz, 2H), 1.04 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 172.81, 152.43, 145.44, 144.32, 144.07, 140.64, 139.90, 138.72, 134.38, 129.13, 129.01, 128.94, 128.69, 115.92, 80.43, 54.92, 41.39, 40.67, 22.17, 17.50, 11.74. 1; LCMS C26H34F3N8O2S2 method (B) Rt=4.102 min, ESI+ m/z=539.3 (M+H).
tert-Butyl (2-((3′-((4-(4,6-diaminopyrimidin-2-yl)thiazol-2-yl)(propyl)amino)-4′-methyl-[1,1′-biphenyl])-4-sulfonamido)ethyl)carbamate OR0651-1 (method A, 29%) as a white powder. Rf=0.6 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.90 (d, J=8.4 Hz, 2H), 7.66 (d, J=8.4 Hz, 2H), 7.51 (dd, J=8.0, 1.7 Hz, 1H), 7.45 (d, J=1.7 Hz, 1H), 7.40 (d, J=8.0 Hz, 1H), 7.33 (s, 1H), 5.77 (brs, 1H), 5.50 (s, 1H), 5.12 (brs, 4H), 5.08 (brs, 1H), 3.93 (brs, 2H), 3.25-3.21 (m, 2H), 3.11-3.05 (m, 2H), 2.24 (s, 3H), 1.62 (sext, 7.3 Hz, 2H), 1.40 (s, 9H), 0.90 (t, J=7.3 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.92, 163.17, 156.74, 144.15, 143.47, 143.21, 139.00, 138.82, 137.80, 132.86, 128.31, 127.80, 127.63, 127.58, 127.25, 111.41, 82.86, 80.00, 54.02, 43.88, 40.48, 28.49, 21.28, 17.63, 11.41; LCMS C30H38N8O4S2 method (B) Rt=4.890 min, ESI+ m/z=639.3 (M+H).
2-(2-((2-Methyl-5-(4-methyl-6-((4-methylpiperazin-1-yl)sulfonyl)pyridin-3-yl)phenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0652 (method B, 31%) as a light yellow powder. Rf=0.23 (DCM-MeOH, 90:10); 1H NMR (400 MHz, CDCl3) δ 8.51 (s, 1H), 7.82 (s, 1H), 7.48 (d, J=7.9 Hz, 1H), 7.42 (s, 1H), 7.28 (dd, J=7.9, 1.4 Hz, 1H), 7.20 (d, J=1.4 Hz, 1H), 5.95 (brs, 4H), 5.69 (s, 1H), 4.04 (brs, 2H), 3.40-3.36 (m, 4H), 2.54-2.50 (m, 4H), 2.38 (s, 3H), 2.31 (s, 3H), 2.30 (s, 3H), 1.67 (sext, J=7.4 Hz, 2H), 0.97 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.90, 161.56, 158.18, 154.50, 150.25, 147.16, 142.97, 140.78, 139.57, 137.91, 136.05, 132.93, 130.55, 129.41, 124.70, 112.64, 82.10, 54.50, 53.80, 46.57, 45.93, 21.40, 20.43, 17.62, 11.50; LCMS C28H35N9O2S2 method (B) Rt=4.434 min, ESI+ m/z=594.2 (M+H).
2-(2-((2-Methyl-5-(2-methyl-6-((4-methylpiperazin-1-yl)sulfonyl)pyridin-3-yl)phenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0653 (method B, 48%) as a light yellow powder. Rf=0.23 (DCM-MeOH, 90:10); 1H NMR (400 MHz, CDCl3) δ 7.80 (d, J=8.0 Hz, 1H), 7.71 (d, J=8.0 Hz, 1H), 7.45 (d, J=7.9 Hz, 1H), 7.36 (s, 1H), 7.24 (dd, J=7.9, 1.8 Hz, 1H), 7.19 (d, J=1.8 Hz, 1H), 5.53 (s, 1H), 5.20 (brs, 4H), 3.99 (brs, 2H), 3.42-3.34 (m, 4H), 2.55 (s, 3H), 2.59-2.51 (m, 4H), 2.30 (s, 3H), 2.29 (s, 3H), 1.65 (sext, J=7.4 Hz, 2H), 0.94 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.87, 163.09, 157.39, 154.07, 149.83, 143.05, 139.16, 138.69, 138.00, 137.84, 132.69, 130.21, 129.04, 120.72, 111.51, 82.70, 54.55, 53.65, 46.76, 46.00, 23.78, 21.37, 17.60, 11.43; LCMS C28H35N9O2S2 method (B) Rt=4.306 min, ESI+ m/z=594.5 (M+H).
2-(2-((5-(6-Methoxy-4-(trifluoromethyl)pyridin-3-yl)-2-methylphenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0654 (method A, 25%) as a light yellow powder. Rf=0.23 (DCM-MeOH, 90:10); 1H NMR (400 MHz, CDCl3) δ 8.17 (s, 1H), 7.39 (d, J=7.9 Hz, 1H), 7.34 (s, 1H), 7.24 (dd, J=7.9, 1.7 Hz, 1H), 7.20 (d, J=1.7 Hz, 1H), 7.07 (s, 1H), 5.52 (s, 1H), 5.10 (brs, 4H), 4.00 (s, 5H), 2.28 (s, 3H), 1.61 (sext, J=7.4 Hz, 2H), 0.92 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 171.18, 164.02, 163.30, 149.81, 142.43, 138.91, 138.60, 137.51, 135.57, 132.00, 131.09, 129.79, 127.29, 124.11, 121.38, 111.49, 107.99, 82.75, 54.18, 53.46, 21.23, 17.54, 11.41; LCMS C24H24F3N7OS method (B) Rt=5.882 min, ESI+ m/z=516.2 (M+H).
2-(2-((5-(6-Hydroxy-4-(trifluoromethyl)pyridin-3-yl)-2-methylphenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0655 (method A, 95%) as a light yellow powder. Rf=0.66 (DCM-MeOH—NH4OH, 80:18:2); 1H NMR (400 MHz, MeOD) δ 7.60 (s, 1H), 7.50 (s, 1H), 7.49 (d, J=7.9 Hz, 1H), 7.34 (dd, J=7.9, 2.0 Hz, 1H), 7.29 (d, J=2.0 Hz, 1H), 6.91 (s, 1H), 5.64 (s, 1H), 4.01 (brs, 2H), 2.30 (s, 3H), 1.70 (sext, J=7.4 Hz, 2H), 1.00 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 172.71, 163.90, 154.10, 145.52, 143.34, 142.64, 138.86, 138.58, 135.95, 133.42, 132.26, 131.93, 124.99, 122.25, 119.06, 118.28, 115.05, 80.92, 54.68, 22.08, 17.47, 11.70; LCMS C23H22F3N7OS method (B) Rt=4.323 min, ESI+ m/z=502.2 (M+H).
2-(2-((2-Methyl-5-(6-((4-methylpiperazin-1-yl)sulfonyl)-4-(trifluoromethyl)pyridin-3-yl)phenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0656 (method B, 59%) as a light yellow powder. Rf=0.23 (DCM-MeOH, 90:10); 1H NMR (400 MHz, MeOD) δ 8.84 (s, 1H), 8.22 (s, 1H), 7.58 (d, J=8.2 Hz, 1H), 7.44 (dd, J=8.2, 1.9 Hz, 1H), 7.42 (d, J=1.9 Hz, 1H), 7.33 (s, 1H), 5.55 (s, 1H), 4.00 (brs, 2H), 3.42-3.36 (m, 4H), 2.56-2.50 (m, 4H), 2.35 (s, 3H), 2.31 (s, 3H), 1.71 (sext, J=7.5 Hz, 2H), 0.98 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 171.98, 164.96, 159.93, 157.81, 154.42, 150.90, 144.11, 140.02, 138.84, 135.45, 133.60, 131.68, 130.57, 125.14, 122.41, 120.14, 111.89, 82.60, 55.42, 54.75, 47.34, 45.89, 22.18, 17.66, 11.59; LCMS C28H32F3N9O2S2 method (B) Rt=4.511 min, ESI+ m/z=648.2 (M+H).
2-(2-((2-Methyl-5-(6-((4-methylpiperazin-1-yl)sulfonyl)-2-(trifluoromethyl)pyridin-3-yl)phenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0657 (method B, 55%) as a light yellow powder. Rf=0.23 (DCM-MeOH, 90:10); 1H NMR (400 MHz, CDCl3) δ 8.13 (d, J=8.0 Hz, 1H), 7.98 (d, J=8.0 Hz, 1H), 7.45 (d, J=7.9 Hz, 1H), 7.37 (s, 1H), 7.26 (dd, J=7.9, 1.9 Hz, 1H), 7.23 (d, J=1.9 Hz, 1H), 5.54 (s, 1H), 5.28 (brs, 4H), 3.99 (brs, 2H), 3.50-3.44 (m, 4H), 2.54-2.48 (m, 4H), 2.31 (s, 3H), 2.30 (s, 3H), 1.62 (sext, J=7.4 Hz, 2H), 0.93 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.83, 162.95, 155.47, 145.45, 145.11, 142.77, 142.57, 139.06, 138.86, 135.16, 132.47, 130.07, 128.75, 124.96, 122.57, 119.82, 111.78, 82.65, 54.56, 53.73, 46.79, 46.03, 21.25, 17.68, 11.42; LCMS C28H32F3N9O2S2 method (B) Rt=4.601 min, ESI+ m/z=648.2 (M+H).
2-(2-((2-Methyl-5-(6-(piperazin-1-ylsulfonyl)-2-(trifluoromethyl)pyridin-3-yl)phenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0658 (method C, 65%) as a light yellow powder. Rf=0.18 (DCM-MeOH, 90:10); 1H NMR (400 MHz, CDCl3) δ 8.12 (d, J=8.1 Hz, 1H), 7.98 (d, J=8.1 Hz, 1H), 7.45 (d, J=7.9 Hz, 1H), 7.36 (s, 1H), 7.26 (dd, J=7.9, 1.9 Hz, 1H), 7.26 (d, J=1.9 Hz, 1H), 5.45 (s, 1H), 5.00 (brs, 4H), 3.97 (brs, 2H), 3.46-3.40 (m, 4H), 3.00-2.94 (m, 4H), 2.31 (s, 3H), 1.62 (sext, J=7.5 Hz, 2H), 0.92 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.81, 163.41, 159.60, 155.57, 150.36, 145.40, 142.82, 142.61, 139.07, 138.90, 135.14, 132.45, 130.08, 128.70, 124.98, 121.22, 111.48, 82.82, 53.74, 47.70, 45.70, 21.21, 17.69, 11.38; LCMS C27H30F3N9O2S2 method (B) Rt=4.308 min, ESI+m/z=634.2 (M+H).
General Procedure for the Synthesis of 2-(2-((5-halogeno-2-methylphenyl)(alkyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine. As previously described for synthesis of 2-[2-(biaryl-3-ylamino)-thiazol-4-yl]-pyrimidine-4,6-diamine derivative, method (A).
2-(2-((5-Iodo-2-methylphenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0607-1 (40%) as a light yellow solid. Rf=0.30 (DCM-MeOH—NH4OH, 95:5:0.5); 1H NMR (400 MHz, CDCl3) δ 7.60 (dd, J=8.1, 1.7 Hz, 1H), 7.55 (d, J=1.7 Hz, 1H), 7.36 (s, 1H), 7.06 (d, J=8.1 Hz, 1H), 5.50 (s, 1H), 5.08 (brs, 4H), 3.97-3.89 (m, 2H), 2.17 (s, 3H), 1.60 (sext, J=7.4 Hz, 2H), 0.91 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.59, 163.34, 159.47, 150.29, 144.20, 138.28, 137.65, 137.58, 133.74, 111.53, 90.93, 82.85, 53.91, 21.22, 17.54, 11.37; LCMS C17H19N6S method (B) Rt=5.226 min, ESI+ m/z=467.0 (M+H).
2-(2-((5-Bromo-2-methylphenyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0614-1 (23%) as a light brown solid. Rf=0.15 (DCM-MeOH—NH4OH, 95:5:0.5); 1H NMR (400 MHz, MeOD) δ 7.87 (d, J=1.8 Hz, 1H), 7.46 (s, 1H), 7.19 (dd, J=8.1, 1.8 Hz, 1H), 7.15 (d, J=8.1 Hz, 1H), 5.54 (s, 1H), 2.27 (s, 3H); LCMS C14H13BrN6S method (B) Rt=4.752 min, ESI+ m/z=337.0 (M+H).
2-(2-((5-Bromo-2-methylphenyl)(propyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0652-1 (36%) as a light yellow solid. Rf=0.35 (DCM-MeOH—NH4OH, 95:5:0.5); 1H NMR (400 MHz, CDCl3) δ 7.41 (dd, J=8.2, 2.0 Hz, 1H), 7.37 (d, J=2.0 Hz, 1H), 7.35 (s, 1H), 7.20 (d, J=8.2 Hz, 1H), 5.46 (s, 1H), 5.01 (brs, 4H), 3.95-3.87 (m, 2H), 2.16 (s, 3H), 1.60 (sext, J=7.4 Hz, 2H), 0.90 (t, J=7.4 Hz, 3H); LCMS C17H19BrN6S method (B) Rt=5.340 min, ESI+m/z=419.0 (M+H). 2-(2-((5-Bromo-2-methylphenyl)(methyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0631-1 (50%) as a light brown solid. Rf=0.60 (DCM-MeOH—NH4OH, 90:10:1); 1H NMR (400 MHz, MeOD) δ 7.51 (d, J=2.0 Hz, 1H), 7.49 (dd, J=8.3, 2.0 Hz, 1H), 7.32 (d, J=8.3 Hz, 1H), 7.30 (s, 1H), 5.53 (s, 1H), 3.51 (s, 3H), 2.22 (s, 3H); LCMS C15H15BrN6S method (B) Rt=5.129 min, ESI+ m/z=391.0 (M+H).
2-(2-((5-Bromo-2-methylphenyl)(ethyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0632-1 (40%) as a light brown solid. Rf=0.45 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 7.49 (dd, J=8.1, 2.0 Hz, 1H), 7.46 (d, J=2.0 Hz, 1H), 7.31 (d, J=8.1 Hz, 1H), 7.29 (s, 1H), 5.53 (s, 1H), 4.04 (q, J=7.1 Hz, 2H), 2.20 (s, 3H), 1.22 (t, J=7.1 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 171.20, 165.46, 160.93, 151.84, 145.16, 138.02, 134.70, 133.57, 132.92, 120.94, 111.38, 82.94, 47.83, 17.37, 13.40; LCMS C16H17BrN6S method (B) R=5.205 min, ESI+ m/z=405.0 (M+H).
2-(2-((5-Bromo-2-methylphenyl)(isopropyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0636-1 (35%) as a light yellow solid. Rf=0.45 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 7.53 (dd, J=8.3, 2.1 Hz, 1H), 7.41 (d, J=2.1 Hz, 1H), 7.35 (d, J=8.3 Hz, 1H), 7.28 (s, 1H), 5.54 (s, 1H), 5.13 (sept, J=6.7 Hz, 1H), 2.21 (s, 3H), 1.24 (d, J=6.7 Hz, 6H); 13C NMR (100 MHz, MeOD) δ 171.34, 165.39, 160.85, 151.69, 142.68, 139.55, 134.95, 134.74, 133.26, 120.67, 111.20, 82.85, 53.00, 21.21, 17.96; LCMS C17H19BrN6S method (B) Rt=5.252 min, ESI+ m/z=419.0 (M+H).
2-(2-((5-Bromo-2-methylphenyl)(isobutyl)amino)thiazol-4-yl)pyrimidine-4,6-diamine OR0637-1 (40%) as a light brown solid. Rf=0.45 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, MeOD) δ 7.51-7.47 (m, 2H), 7.32 (d, J=8.8 Hz, 1H), 7.28 (s, 1H), 5.53 (s, 1H), 3.84 (brs, 2H), 2.21 (s, 3H), 2.02-1.89 (m, 1H), 1.01 (d, J=6.7 Hz, 6H); 13C NMR (100 MHz, MeOD) δ 172.24, 165.59, 161.19, 152.04, 146.07, 137.69, 135.00, 133.30, 132.76, 120.84, 111.40, 82.98, 60.56, 28.64, 20.71, 17.59; LCMS C18H21BrN6S method (B) Rt=5.382 min, ESI+ m/z=433.0 (M+H).
General Procedure for the Synthesis of ethyl 2-([1,1′-biaryl]-3-ylamino)thiazole-4-carboxylate derivatives. Method (A): as previously described for dCKi-2-1, to a stirred solution of appropriate 1-([1,1′-biaryl]-3-yl)thiourea (10.0 mmol) in ethanol (100 mL) was added ethyl bromopyruvate (1.5 mL, 12.0 mmol). The resulting mixture was heated at reflux for 3 hrs, then allowed to cool to room temperature and concentrated under reduced pressure. The residue was purified by flash chromatography to afford expected ethyl 2-([1,1′-biaryl]-3-ylamino)thiazole-4-carboxylate derivative. Method (B): under argon, a suspension of appropriate aryl halide (1 mmol), ethyl 2-((5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)amino) thiazole-4-carboxylate or analogue (1.1 mmol), PdCl2(dppf) (36 mg, 0.05 mmol) and Na2CO3 (212 mg, 2 mmol) in a degassed mixture of 1,4-dioxane-water (5:1, 12 mL) was refluxed for 2 hrs, upon complete consumption of starting material. The solvent was distillated off under reduced pressure and the residue purified by flash chromatography to afford corresponding ethyl 2-([1,1′-biaryl]-3-ylamino)thiazole-4-carboxylate derivative or analogue. Method (C): under argon, to a solution of 4-halogenoarylsulfonyl chloride (1.0 mmol) in 1,4-dioxane (12 mL) were added successively appropriate amine (1.15 mmol) and K2CO3 (3.0 mmol). The reaction mixture was stirred at room temperature until complete consumption of starting material monitored by LCMS. To the resulting suspension, were added successively ethyl 2-((2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)(propyl)amino)thiazole-4-carboxylate OR0625-2 (1.10 mmol), PdCl2(dppf) (0.15 mmol), and water (2 mL). The reaction mixture was thoroughly degassed several times under argon fillings, and then heated at 80° C. for 2 hours. The solvent was distillated off and the residue was triturated with DCM (3×50 mL). The combined organic layers were dried over Na2SO4. The solvent was distillated off under reduced pressure, and the residue was purified by flash chromatography to afford expected ethyl 2-([1,1′-biaryl]-3-ylamino)thiazole-4-carboxylate.
Ethyl 2-((4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazole-4-carboxylate OR0600-1 (method A, 86%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 7.53 (dd, J=8.0, 1.9 Hz, 1H), 7.49 (d, J=8.2 Hz, 2H), 7.43 (d, J=1.9 Hz, 1H), 7.40 (d, J=8.0 Hz, 1H), 7.29 (s, 1H), 7.28 (d, J=8.2 Hz, 2H), 4.36 (q, J=7.1 Hz, 2H), 3.85 (brs, 2H), 2.88-2.80 (m, 2H), 2.67-2.59 (m, 2H), 2.50 (brs, 8H), 2.31 (s, 3H), 2.25 (s, 3H), 1.69 (sext, J=7.4 Hz, 2H), 1.38 (t, J=7.1 Hz, 3H), 0.94 (t, J=7.4 Hz, 3H); LCMS C29H38N4O2S method (B) Rc=5.102 min. ESI+ m/z=507.3 (M+H).
2-((4-Methyl-4′-(3-(4-methylpiperazin-1-yl)propyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazole-4-carboxylate OR0601-1 (method A, 93%) as a beige foam. Rf=0.35 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.53 (dd, J=7.9, 1.9 Hz, 1H), 7.48 (d, J=8.1 Hz, 2H), 7.42 (d, J=1.9 Hz, 1H), 7.39 (d, J=7.9 Hz, 1H), 7.28 (s, 1H), 7.24 (d, J=8.1 Hz, 2H), 4.35 (q, J=7.1 Hz, 2H), 3.93 (brs, 2H), 2.73 (brs, 8H), 2.72-2.64 (m, 2H), 2.54-2.46 (m, 2H), 2.44 (s, 3H), 2.25 (s, 3H), 1.90 (quint, J=7.6 Hz, 2H), 1.69 (sext, J=7.4 Hz, 2H), 1.37 (t, J=7.1 Hz, 3H), 0.94 (t, J=7.4 Hz, 3H). 13C NMR (100 MHz, CDCl3) δ 170.89, 162.09, 143.97, 143.18, 141.20, 140.96, 137.65, 135.87, 132.68, 129.06, 127.83, 127.24, 126.97, 116.75, 61.11, 57.47, 54.30, 53.68, 51.98, 45.31, 33.16, 28.02, 21.20, 17.37, 14.48, 11.43; LCMS C30H40N4O2S method (B) Rt=5.158 min, ESI+ m/z=521.4 (M+H).
Ethyl 2-((2-methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethyl)pyridin-3-yl)phenyl)(propyl)amino)thiazole-4-carboxylate OR0602-1 (method A, 47%) as a light brown oil. LCMS C28H37N5O2S method (B) Rt=4.730 min, ESI+ m/z=508.2 (M+H).
Ethyl 2-(isobutyl(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)thiazole-4-carboxylate OR0603-1 (method A, 57%) as a light brown solid. LCMS C30H40N4O2S method (B) Rt=5.363 min, ESI+ m/z=521.2 (M+H).
Ethyl 2-((cyclopropylmethyl)(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)thiazole-4-carboxylate OR0604-1 (method A, 100%) as a yellow solid. LCMS C30H38N4O2S method (B) Rt=5.138 min, ESI+ m/z=519.3 (M+H).
Ethyl 2-(iso-pentyl(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)thiazole-4-carboxylate OR0605-1 (method A, 31%) as a light yellow solid. Rf=0.45 (DCM-MeOH—NH4OH, 90:9:1); LCMS C31H42N4O2S method (B) Rt=5.460 min, ESI+m/z=535.3 (M+H).
Ethyl 2-(butyl(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)thiazole-4-carboxylate OR0606-1 (method A, 89%), a light yellow solid, engaged in the next step without further purification. Rf=0.45 (DCM-MeOH, 90:10); 1H NMR (400 MHz, CDCl3) δ 7.53 (dd, J=8.0, 1.9 Hz, 1H), 7.49 (d, J=8.2 Hz, 2H), 7.42 (d, J=1.9 Hz, 1H), 7.40 (d, J=8.0 Hz, 1H), 7.28 (s, 1H), 7.28 (d, J=8.2 Hz, 2H), 4.35 (q, J=7.1 Hz, 2H), 3.96 (brs, 2H), 2.90-2.82 (m, 2H), 2.72-2.64 (m, 2H), 2.58 (brs, 8H), 2.37 (s, 3H), 2.25 (s, 3H), 1.63 (quint, J=7.8 Hz, 2H), 1.42-1.34 (m, 2H), 1.38 (t, J=7.1 Hz, 2H), 0.91 (t, J=7.4 Hz, 3H). 13C NMR (100 MHz, CDCl3) δ 170.89, 162.11, 143.97, 143.20, 140.95, 139.72, 137.86, 135.94, 132.70, 129.40, 127.87, 127.26, 127.01, 116.75, 61.12, 60.21, 54.93, 52.71, 51.88, 45.79, 33.18, 30.06, 20.26, 17.40, 14.49, 14.04; LCMS C30H40N4O2S method (B) Rt=5.117 min, ESI+ m/z=521.3 (M+H).
Ethyl 2-(isobutyl(2-methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethoxy)pyridin-3-yl)phenyl)amino)thiazole-4-carboxylate OR0608-1 (method A, 100%) as a light brown solid. LCMS C29H39N5O3S method (B) Rt=5.274 min, ESI+ m/z=538.2 (M+H).
Ethyl 2-(isobutyl(2-methyl-5-(4-(methylsulfonamidomethyl)-1H-1,2,3-triazol-1-yl)phenyl)amino)thiazole-4-carboxylate OR0609-1 (method A, 38%) as a yellow solid. 1H NMR (400 MHz, CDCl3) δ 8.05 (s, 1H), 7.68 (d, J=2.0 Hz, 1H), 7.64 (dd, J=8.3, 2.0 Hz, 1H), 7.49 (d, J=8.3 Hz, 1H), 7.34 (s, 1H), 5.22 (brs, 1H), 4.52 (s, 2H), 4.34 (q, J=7.1 Hz, 2H), 3.81 (brs, 2H), 3.01 (s, 3H), 2.29 (s, 3H), 2.06-2.00 (m, 1H), 1.36 (t, J=7.1 Hz, 3H), 0.99 (d, J=6.8 Hz, 6H); LCMS C21H28N6O4S2 method (B)Rt=6.189 min, ESI+ m/z=493.2 (M+H).
Ethyl 2-((2-((tert-butoxycarbonyl)amino)ethyl)(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)thiazole-4-carboxylate OR0610-2 (method A, 399 mg, 74%) as a light yellow solid. Rf=0.47 (DCM-MeOH, 90:10); 1H NMR (400 MHz, CDCl3) δ 7.53 (dd, J=7.9, 1.8 Hz, 1H), 7.49 (d, J=8.2 Hz, 2H), 7.48 (d, J=1.8 Hz, 1H), 7.38 (d, J=7.9 Hz, 1H), 7.30 (s, 1H), 7.25 (d, J=8.2 Hz, 2H), 5.56 (brs, 1H), 4.34 (q, J=7.1 Hz, 2H), 3.55-3.47 (m, 2H), 2.88-2.80 (m, 2H), 2.69-2.61 (m, 2H), 2.58 (brs, 8H), 2.35 (s, 3H), 2.23 (s, 3H), 1.37 (t, J=7.1 Hz, 3H), 1.35 (s, 9H); LCMS C33H45N5O4S method (B) Rt=5.147 min, ESI+ m/z=608.3 (M+H).
Ethyl 2-((3-((tert-butoxycarbonyl)amino)propyl)(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)thiazole-4-carboxylate OR0611-2 (method A, 67%) as a pale yellow solid. Rf=0.55 (DCM-MeOH, 90:10); 1H NMR (400 MHz, CDCl3) δ 7.54 (dd, J=8.0, 1.8 Hz, 1H), 7.47 (d, J=8.2 Hz, 2H), 7.40 (d, J=8.0 Hz, 1H), 7.36 (d, J=1.8 Hz, 1H), 7.29 (s, 1H), 7.27 (d, J=8.2 Hz, 2H), 6.25 (brs, 1H), 4.39 (q, J=6.9 Hz, 2H), 4.29 (brs, 1H), 3.70 (brs, 1H), 3.42-3.34 (m, 2H), 3.28-3.20 (m, 2H), 2.89-2.81 (m, 2H), 2.71-2.63 (m, 2H), 2.62 (brs, 8H), 2.39 (s, 3H), 2.24 (s, 3H), 1.83-1.75 (m, 2H), 1.46 (s, 9H), 1.39 (t, J=6.9 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 171.27, 161.95, 156.50, 143.61, 143.10, 141.30, 139.72, 137.70, 135.42, 132.93, 129.39, 127.90, 127.58, 127.05, 116.80, 78.90, 61.14, 60.08, 54.79, 52.51, 49.56, 45.64, 37.64, 33.10, 28.95, 28.65, 17.31, 14.61; LCMS C34H47N5O4S method (B) Rt=5.401 min, ESI+ m/z=622.4 (M+H).
Ethyl 2-((2-methyl-5-(6-(4-methylpiperazine-1-carbonyl)pyridin-3-yl)phenyl)(propyl)amino)thiazole-4-carboxylate OR0612-1 (method A, 62%) as a light yellow oil. 1H NMR (400 MHz, CDCl3) δ 8.78 (d, J=2.0 Hz, 1H), 7.97 (dd, J=8.2, 2.0 Hz, 1H), 7.76 (d, J=8.2 Hz, 1H), 7.56 (dd, J=8.0, 1.9 Hz, 1H), 7.49 (d, J=8.0 Hz, 1H), 7.46 (d, J=1.9 Hz, 1H), 7.30 (s, 1H), 4.36 (q, J=7.1 Hz, 2H), 3.94 (brs, 4H), 3.81 (brs, 2H), 2.68 (brs, 2H), 2.60 (brs, 2H), 2.44 (s, 3H), 2.29 (s, 3H), 1.70 (sext, J=7.4 Hz, 2H), 1.38 (t, J=7.1 Hz, 3H), 0.96 (t, J=7.4 Hz, 3H); LCMS C27H33N5O3S method (B) Rt=5.007 min, ESI+ m/z=508.2 (M+H).
Ethyl 2-((2-methyl-5-(6-(4-methylpiperazine-1-carboxamido)pyridin-3-yl)phenyl)(propyl)amino)thiazole-4-carboxylate OR0616-1 (method A, 45%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 8.39 (d, J=2.3 Hz, 1H), 8.07 (d, J=8.7 Hz, 1H), 7.85 (dd, J=8.7, 2.3 Hz, 1H), 7.50 (dd, J=8.0, 1.9 Hz, 1H), 7.43 (d, J=8.0 Hz, 1H), 7.40 (d, J=1.9 Hz, 1H), 7.30 (s, 1H), 4.35 (q, J=7.1 Hz, 2H), 3.91 (brs, 2H), 3.75 (brs, 4H), 2.73 (brs, 4H), 2.51 (s, 3H), 2.26 (s, 3H), 1.69 (sext, J=7.4 Hz, 2H), 1.37 (t, J=7.1 Hz, 3H), 0.95 (t, J=7.4 Hz, 3H); LCMS C27H34N6O3S method (B) Rt=5.089 min, ESI+ m/z=523.3 (M+H).
Ethyl 2-((2-methyl-5-(6-((4-methylpiperazine)-1-sulfonamido)pyridin-3-yl)phenyl)(propyl)amino)thiazole-4-carboxylate OR0625-1 (method B, 41%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 8.34 (d, J=2.2 Hz, 1H), 7.88 (dd, J=9.1, 2.2 Hz, 1H), 7.54 (d, J=9.1 Hz, 1H), 7.47 (dd, J=8.0, 1.7 Hz, 1H), 7.44 (d, J=8.0 Hz, 1H), 7.36 (d, J=1.7 Hz, 1H), 7.31 (s, 1H), 4.35 (q, J=7.1 Hz, 2H), 3.91 (brs, 2H), 3.53 (brs, 4H), 2.91 (brs, 4H), 2.56 (s, 3H), 2.27 (s, 3H), 1.69 (sext, J=7.4 Hz, 2H), 1.38 (t, J=7.1 Hz, 3H), 0.95 (t, J=7.4 Hz, 3H); LCMS C26H34N6O4S2 method (B) Rt=5.084 min, ESI+ m/z=559.3 (M+H).
Ethyl 2-((2-methyl-5-(6-((4-methylpiperazin-1-yl)sulfonyl)pyridin-3-yl)phenyl)(propyl)amino)thiazole-4-carboxylate OR0627-1 (method B, 68%) as a light brown solid. Rf=0.50 (DCM-MeOH, 95:5); 1H NMR (400 MHz, CDCl3) δ 8.88 (d, J=2.2 Hz, 1H), 8.04 (dd, J=8.1, 2.2 Hz, 1H), 7.97 (d, J=8.1 Hz, 1H), 7.56 (dd, J=7.9, 1.9 Hz, 1H), 7.51 (d, J=7.9 Hz, 1H), 7.46 (d, J=1.9 Hz, 1H), 7.31 (s, 1H), 4.36 (q, J=7.1 Hz, 2H), 3.94 (brs, 2H), 3.46 (brs, 4H), 2.63 (brs, 4H), 2.39 (s, 3H), 2.30 (s, 3H), 1.70 (sext, J=7.4 Hz, 2H), 1.38 (t, J=7.1 Hz, 3H), 0.96 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.37, 161.98, 154.68, 148.45, 144.14, 143.85, 138.78, 138.61, 136.00, 135.83, 133.53, 128.51, 127.59, 123.26, 116.90, 61.21, 54.34, 53.90, 46.13, 24.99, 21.27, 17.63, 14.49, 11.43; LCMS C26H33N5O4S2 method (B) Rt=5.281 min, ESI+ m/z=544.2 (M+H).
Ethyl 2-((2-methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethoxy)pyridin-3-yl)phenyl)(propyl)amino)thiazole-4-carboxylate OR0629-1 (method B, 84%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 8.33 (d, J=2.3 Hz, 1H), 7.75 (dd, J=8.7, 2.3 Hz, 1H), 7.47 (dd, J=7.9, 1.9 Hz, 1H), 7.41 (d, J=7.9 Hz, 1H), 7.37 (d, J=1.9 Hz, 1H), 7.29 (s, 1H), 6.82 (d, J=8.7 Hz, 1H), 4.48 (t, J=5.8 Hz, 2H), 4.35 (q, J=7.1 Hz, 2H), 3.89 (brs, 2H), 2.84 (t, J=5.8 Hz, 2H), 2.71 (brs, 4H), 2.62 (brs, 4H), 2.37 (s, 3H), 2.25 (s, 3H), 1.69 (sext, J=7.4 Hz, 2H), 1.37 (t, J=7.1 Hz, 3H), 0.94 (t, J=7.4 Hz, 3H); LCMS C28H37N5O3S method (B) Rt=5.247 min, ESI+ m/z=524.3 (M+H).
Ethyl 2-((2-methyl-5-(6-(2-(4-methylpiperazin-1-yl)-2-oxoethoxy)pyridin-3-yl)phenyl)(propyl)amino)thiazole-4-carboxylate OR0630-1 (method B, 71%) as a light brown oil. 1H NMR (400 MHz, CDCl3) δ 8.29 (d, J=2.4 Hz, 1H), 7.79 (dd, J=8.6, 2.4 Hz, 1H), 7.47 (dd, J=8.0, 1.8 Hz, 1H), 7.41 (d, J=8.0 Hz, 1H), 7.36 (d, J=1.8 Hz, 1H), 7.29 (s, 1H), 6.96 (d, J=8.6 Hz, 1H), 5.05 (s, 2H), 4.35 (q, J=7.1 Hz, 2H), 3.91 (brs, 2H), 3.74 (brs, 2H), 3.63 (brs, 2H), 2.62-2.50 (m, 4H), 2.40 (s, 3H), 2.25 (s, 3H), 1.69 (sext, J=7.4 Hz, 2H), 1.37 (t, J=7.1 Hz, 3H), 0.94 (t, J=7.4 Hz, 3H); LCMS C28H35N5O4S method (B) Rt=5.234 min, ESI+ m/z=538.3 (M+H).
Ethyl 2-((5-fluoro-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazole-4-carboxylate OR0633-1 (method A, 86%) as a yellow oil. LCMS C27H33FN4O4S2 method (B) Rt=5.584 min, ESI+ m/z=561.2 (M+H).
Ethyl 2-((3′-methoxy-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazole-4-carboxylate OR0635-1 (method B, 82%) as a light brown solid. Rf=0.50 (DCM-MeOH, 95:5); 1H NMR (400 MHz, CDCl3) δ 7.91 (d, J=8.0 Hz, 1H), 7.55 (dd, J=8.0, 1.9 Hz, 1H), 7.45 (d, J=8.2 Hz, 1H), 7.44 (d, J=1.9 Hz, 1H), 7.31 (s, 1H), 7.20 (dd, J=8.2, 1.4 Hz, 1H), 7.12 (d, J=1.4 Hz, 1H), 4.36 (q, J=7.1 Hz, 2H), 3.99 (s, 3H), 3.97 (brs, 2H), 3.44 (brs, 4H), 2.70 (brs, 4H), 2.46 (s, 3H), 2.29 (s, 3H), 1.69 (sext, J=7.4 Hz, 2H), 1.38 (t, J=7.1 Hz, 3H), 0.96 (t, J=7.4 Hz, 3H); LCMS C28H36FN4O5S2 method (B) R=5.258 min, ESI+ m/z=574.3 (M+H).
Ethyl 2-((4′-((4-ethylpiperazin-1-yl)sulfonyl)-4-methyl-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazole-4-carboxylate OR0638-1 (method B, 82%) as a colorless oil. LCMS C28H36N4O4S2 method (B) Rt=5.183 min, ESI+ m/z=557.4 (M+H).
Ethyl 2-((4′-((4-(tert-butoxycarbonyl)piperazin-1-yl)sulfonyl)-4-methyl-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazole-4-carboxylate OR0639-2 (method B, 56%) as a white solid. LCMS C31H40N4O6S2 method (B) Rt=7.658 min.
Ethyl 2-((4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)oxazole-4-carboxylate OR0640-1 (method B, 78%) as a light brown solid. 1H NMR (400 MHz, CDCl3) δ 7.80 (d, J=8.5 Hz, 2H), 7.71 (d, J=8.5 Hz, 2H), 7.69 (s, 1H), 7.50 (dd, J=7.8, 1.8 Hz, 1H), 7.41 (d, J=7.8 Hz, 1H), 7.40 (d, J=1.8 Hz, 1H), 4.35 (q, J=7.1 Hz, 2H), 3.87-3.79 (m, 2H), 3.18 (brs, 4H), 2.66 (brs, 4H), 2.40 (s, 3H), 2.24 (s, 3H), 1.70 (sext, J=7.4 Hz, 2H), 1.36 (t, J=7.1 Hz, 3H), 0.94 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 182.08, 162.14, 160.86, 151.89, 145.03, 140.97, 139.05, 138.45, 138.17, 137.34, 134.13, 133.42, 132.38, 128.48, 127.77, 127.61, 126.81, 61.07, 53.95, 53.70, 45.40, 25.00, 21.20, 17.68, 14.49, 11.30; LCMS C27H34N4O5S method (B) Rt=5.157 min, ESI+ m/z=527.3 (M+H).
Ethyl 2-((2′-chloro-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazole-4-carboxylate OR0641-1 (method B, 73%) as a light brown solid. Rf=0.50 (DCM-MeOH, 95:5); 1H NMR (400 MHz, CDCl3) δ 7.85 (d, J=1.8 Hz, 1H), 7.67 (dd, J=8.0, 1.8 Hz, 1H), 7.50 (d, J=8.0 Hz, 1H), 7.46 (d, J=7.9 Hz, 1H), 7.37 (dd, J=7.9, 1.8 Hz, 1H), 7.33 (d, J=1.8 Hz, 1H), 7.30 (s, 1H), 4.35 (q, J=7.1 Hz, 2H), 3.93 (brs, 2H), 3.13-3.11 (m, 4H), 2.55-2.54 (m, 4H), 2.31 (s, 3H), 2.29 (s, 3H), 1.68 (sext, J=7.4 Hz, 2H), 1.37 (t, J=7.1 Hz, 3H), 0.95 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.64, 162.03, 144.03, 143.81, 142.63, 137.88, 137.41, 135.82, 133.75, 132.47, 131.86, 130.68, 129.53, 129.39, 126.31, 116.90, 61.15, 54.02, 53.66, 45.93, 45.67, 21.18, 17.56, 14.48, 11.43; LCMS C27H33ClN4O4S2 method (B) Rt=5.248 min, ESI+ m/z=577.2 (M+H).
Ethyl 2-((4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-2′-(trifluoromethyl)-[1,1′-biphenyl]-3-yl)(propyl)amino) thiazole-4-carboxylate OR0642-1 (method B, 75%) as a light brown solid. Rf=0.65 (DCM-MeOH, 95:5); 1H NMR (400 MHz, CDCl3) δ 8.10 (d, J=1.6 Hz, 1H), 7.93 (dd, J=8.0, 1.6 Hz, 1H), 7.54 (d, J=8.0 Hz, 1H), 7.44 (d, J=7.9 Hz, 1H), 7.31 (s, 1H), 7.28 (dd, J=7.9, 1.4 Hz, 1H), 7.20 (d, J=1.4 Hz, 1H), 4.35 (q, J=7.1 Hz, 2H), 3.93 (brs, 2H), 3.16-3.13 (m, 4H), 2.59-2.56 (m, 4H), 2.34 (s, 3H), 2.29 (s, 3H), 1.66 (sext, J=7.4 Hz, 2H), 1.37 (t, J=7.1 Hz, 3H), 0.94 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.66, 162.04, 144.82, 144.01, 142.40, 137.96, 137.71, 135.41, 133.15, 132.28, 130.68, 129.99, 129.67, 129.01, 125.89, 116.98, 61.16, 53.97, 53.61, 45.82, 45.62, 21.13, 17.54, 14.48, 11.42; LCMS C28H33F3N4O4S2 method (B) Rt=5.454 min, ESI+ m/z=611.3 (M+H).
Ethyl 2-((2′-fluoro-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazole-4-carboxylate OR0643-1 (method B, 62%) as a light brown solid. Rf=0.50 (DCM-MeOH, 95:5); 1H NMR (400 MHz, CDCl3) δ 7.61-7.60 (m, 2H), 7.53-7.51 (m, 2H), 7.47 (d, J=8.0 Hz, 1H), 7.44 (s, 1H), 7.31 (s, 1H), 4.36 (q, J=7.1 Hz, 2H), 3.93 (brs, 2H), 3.23-3.20 (m, 4H), 2.69-2.67 (m, 4H), 2.42 (s, 3H), 2.29 (s, 3H), 1.70 (sext, J=7.4 Hz, 2H), 1.38 (t, J=7.1 Hz, 3H), 0.96 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.61, 162.05, 148.86, 144.08, 143.16, 138.30, 136.18, 133.69, 132.82, 131.46, 130.32, 129.31, 123.89, 116.92, 116.25, 115.99, 61.18, 53.90, 53.78, 53.57, 45.35, 21.20, 17.62, 14.50, 11.44; LCMS C27H33FN4O4S2 method (B) Rt=5.220 min, ESI+ m/z=561.2 (M+H).
Ethyl 2-((2′,6′-difluoro-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazole-4-carboxylate OR0644-1 (method B, 64%) as a light yellow solid. Rf=0.28 (DCM-MeOH, 98:2); 1H NMR (400 MHz, CDCl3) δ 7.48 (d, J=8.0 Hz, 1H), 7.43 (d, J=8.0 Hz, 1H), 7.41-7.32 (m, 3H), 7.31 (s, 1H), 4.35 (q, J=7.1 Hz, 2H), 3.91 (brs, 2H), 3.16-3.13 (m, 4H), 2.56-2.54 (m, 4H), 2.32 (s, 3H), 2.30 (s, 3H), 1.68 (sext, J=7.4 Hz, 2H), 1.37 (t, J=7.1 Hz, 3H), 0.94 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.61, 162.04, 161.10, 158.57, 144.01, 142.90, 138.76, 136.73, 132.58, 131.60, 130.51, 127.08, 117.00, 111.60, 61.16, 54.00, 53.67, 45.68, 44.61, 21.12, 17.64, 14.47, 11.40; LCMS C27H32F2N4O4S2 method (B) Rt=5.283 min, ESI+ m/z=579.2 (M+H).
Ethyl 2-((2′-methoxy-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazole-4-carboxylate OR0645-1 (method B, 55%) as a light brown solid. Rf=0.35 (DCM-MeOH, 95:5); LCMS C28H36N4O5S2 method (B) Rt=5.291 min, ESI+ m/z=573.2 (M+H).
Ethyl 2-((4-methyl-4′-(piperidin-4-ylsulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazole-4-carboxylate OR0646-1 (method B, 70%) as a light brown solid. Rf=0.25 (DCM-MeOH, 95:5); 1H NMR (400 MHz, CDCl3) δ 7.92 (d, J=8.5 Hz, 2H), 7.74 (d, J=8.5 Hz, 2H), 7.58 (dd, J=8.0, 1.9 Hz, 1H), 7.48 (d, J=1.9 Hz, 1H), 7.47 (d, J=8.0 Hz, 1H), 7.30 (s, 1H), 4.35 (q, J=7.1 Hz, 2H), 3.95 (brs, 2H), 3.30-3.24 (m, 2H), 3.12-3.02 (m, 1H), 2.70-2.62 (m, 2H), 2.29 (s, 3H), 2.13-2.05 (m, 2H), 1.76-1.65 (m, 4H), 1.38 (t, J=7.1 Hz, 3H), 0.96 (t, J=7.4 Hz, 3H). 13C NMR (100 MHz, CDCl3) δ 170.56, 162.03, 145.57, 144.10, 143.58, 139.02, 138.09, 135.40, 133.19, 129.96, 128.48, 127.74, 127.67, 116.84, 61.34, 61.21, 53.88, 44.70, 25.33, 21.25, 17.57, 14.49, 11.45; LCMS C27H33N3O4S2 method (B) Rt=5.135 min, ESI+ m/z=528.2 (M+H).
Ethyl 2-((4′-((4-((tert-butoxycarbonyl)amino)piperidin-1-yl)sulfonyl)-4-methyl-[1,1′-biphenyl]-3-yl)(propyl)amino) thiazole-4-carboxylate OR0648-2 (method B, 88%) as a colorless oil. Rf=0.50 (DCM-MeOH, 98:2); 1H NMR (400 MHz, CDCl3) δ 7.81 (d, J=8.3 Hz, 2H), 7.71 (d, J=8.3 Hz, 2H), 7.58 (dd, J=8.0, 1.5 Hz, 1H), 7.48 (d, J=1.5 Hz, 1H), 7.46 (d, J=8.0 Hz, 1H), 7.30 (s, 1H), 4.41 (brs, 1H), 4.36 (q, J=7.1 Hz, 2H), 3.95 (brs, 2H), 3.73 (brs, 1H), 3.39 (brs, 1H), 2.52-2.43 (m, 2H), 2.29 (s, 3H), 2.02-1.95 (m, 2H), 1.71 (sext, J=7.4 Hz, 2H), 1.53-1.47 (m, 2H), 1.40 (s, 9H), 1.37 (t, J=7.1 Hz, 3H), 0.96 (t, J=7.4 Hz, 3H); LCMS C32H42N4O6S2 method (B) Rt=7.522 min, ESI+ m/z=643.2 (M+H).
2-(4-((3′-((4-(Ethoxycarbonyl)thiazol-2-yl)(propyl)amino)-4′-methyl-[1,1′-biphenyl]-4-yl)sulfonyl)piperazin-1-yl)acetic acid OR0649-1 (method B, 42%) as a white solid. Rf=0.11 (DCM-MeOH, 95:5); 1H NMR (400 MHz, MeOD) δ 7.91 (d, J=8.8 Hz, 2H), 7.87 (d, J=8.8 Hz, 2H), 7.73 (dd, J=8.0, 1.9 Hz, 1H), 7.64 (d, J=1.9 Hz, 1H), 7.56 (d, J=8.0 Hz, 1H), 7.45 (s, 1H), 4.33 (q, J=7.1 Hz, 2H), 3.91 (brs, 2H), 3.36 (s, 2H), 3.21 (brs, 4H), 3.03 (brs, 4H), 2.29 (s, 3H), 1.74 (sext, J=7.4 Hz, 2H), 1.36 (t, J=7.1 Hz, 3H), 0.98 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, MeOD) δ 172.17, 163.30, 145.93, 144.85, 144.66, 140.42, 138.85, 135.45, 134.29, 129.78, 129.26, 128.78, 128.72, 118.30, 62.13, 59.86, 55.21, 53.01, 45.86, 22.14, 17.52, 14.60, 11.55; LCMS C28H34N4O6S2 method (B) Rt=5.694 min, ESI+ m/z=588.3 (M+H).
Ethyl 2-((4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-3′-(trifluoromethoxy)-[1,1′-biphenyl]-3-yl)(propyl)amino) thiazole-4-carboxylate OR0650-1 (method B, 82%) as a white solid. Rf=0.36 (DCM-MeOH, 95:5); 1H NMR (400 MHz, CDCl3) δ 8.00 (d, J=8.2 Hz, 1H), 7.56 (dd, J=8.2, 1.6 Hz, 1H), 7.54-7.52 (m, 1H), 7.53 (dd, J=8.0, 2.0 Hz, 1H), 7.48 (d, J=8.0 Hz, 1H), 7.43 (d, J=2.0 Hz, 1H), 7.31 (s, 1H), 4.35 (q, J=7.1 Hz, 2H), 3.94 (brs, 2H), 3.28 (brs, 4H), 2.50 (brs, 4H), 1.68 (sext, J=7.4 Hz, 2H), 1.37 (t, J=7.1 Hz, 3H), 0.95 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.41, 161.96, 146.72, 146.54, 144.11, 143.67, 138.66, 137.98, 133.32, 132.53, 128.87, 128.33, 127.50, 124.95, 121.64, 119.17, 116.87, 61.17, 54.49, 53.89, 45.86, 45.67, 21.23, 17.58, 14.46, 11.40; LCMS C28H33F3N4O5S2 method (B) Rt=5.512 min, ESI+ m/z=627.2 (M+H).
Ethyl 2-((4′—(N-(2-((tert-butoxycarbonyl)amino)ethyl)sulfamoyl)-4-methyl-[1,1′-biphenyl]-3-yl)(propyl)amino)thiazole-4-carboxylate OR0651-2 (method B, 66%) as a colorless oil. Rf=0.33 (DCM-MeOH, 95:5); 1H NMR (400 MHz, CDCl3) δ 7.92 (d, J=8.4 Hz, 2H), 7.68 (d, J=8.4 Hz, 2H), 7.57 (dd, J=7.8, 1.5 Hz, 1H), 7.47 (d, J=1.5 Hz, 1H), 7.46 (d, J=7.8 Hz, 1H), 7.30 (s, 1H), 5.40 (brs, 1H), 4.91 (brs, 1H), 4.35 (q, J=7.1 Hz, 2H), 3.95 (brs, 2H), 3.28-3.20 (m, 2H), 3.14-3.06 (m, 2H), 2.28 (s, 3H), 1.70 (sext, J=7.4 Hz, 2H), 1.41 (s, 9H), 1.37 (t, J=7.1 Hz, 3H), 0.95 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.61, 161.99, 156.74, 144.22, 143.98, 143.48, 139.28, 138.92, 137.72, 133.12, 128.32, 127.84, 127.65, 127.62, 116.82, 80.11, 61.22, 53.91, 43.97, 40.45, 28.46, 21.25, 17.53, 14.49, 11.44; LCMS C29H38N4O6S2 method (B) Rt=7.124 min, ESI+ m/z=603.3 (M+H).
General Procedure for the Synthesis of 1-([1,1′-biaryl]-3-yl)thiourea Derivatives. As previously described above for 1-(3-nitroaryl)thiourea derivatives.
1-iso-Butyl-1-(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)thiourea OR0603-2 (95%) as a white solid. LCMS C25H36N4S method (B) Rt=4.591 min, ESI+ m/z=425.3 (M+H).
1-(Cyclopropylmethyl)-1-(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)thiourea OR0604-2 (70%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 7.53 (dd, J=8.0, 1.9 Hz, 1H), 7.47 (d, J=8.1 Hz, 2H), 7.46 (d, J=1.9 Hz, 1H), 7.37 (d, J=8.0 Hz, 1H), 7.29 (d, J=8.1 Hz, 2H), 5.52 (brs, 2H), 4.38-4.32 (m, 1H), 3.82-3.74 (m, 1H), 2.86-2.82 (m, 2H), 2.67-2.61 (m, 2H), 2.50 (brs, 8H), 2.31 (s, 3H), 2.29 (s, 3H), 1.20-1.14 (m, 1H), 0.49-0.43 (m, 2H), 0.22-0.16 (m, 2H); LCMS C25H34N4S method (B) Rt=4.413 min, ESI+ m/z=423.2 (M+H).
1-iso-Pentyl-1-(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)thiourea OR0605-2 (92%) as a light yellow solid, used in the next step without further purification. Rf=0.18 (DCM-MeOH—NH4OH, 90:9:1); LCMS C26H38N4S method (B) Rt=4.820 min, ESI+ m/z=439.3 (M+H).
1-Butyl-1-(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)thiourea OR0606-2 (63%) as a light yellow solid. Rf=0.12 (DCM-MeOH, 95:5); 1H NMR (400 MHz, CDCl3) δ 7.53 (d, J=7.9 Hz, 1H), 7.47 (d, J=7.9 Hz, 2H), 7.38 (d, J=7.9 Hz, 1H), 7.33 (s, 1H), 7.29 (d, J=7.9 Hz, 2H), 5.51 (brs, 2H), 4.50-4.47 (m, 1H), 3.69-3.67 (m, 1H), 2.87-2.85 (m, 2H), 2.69 (brs, 8H), 2.66-2.64 (m, 2H), 2.38 (s, 3H), 2.26 (s, 3H), 1.73-1.71 (m, 1H), 1.60-1.58 (m, 1H), 1.36-1.35 (m, 2H), 0.91 (t, J=7.3 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 181.86, 140.93, 140.19, 140.04, 137.31, 134.50, 132.64, 129.51, 127.59, 126.98, 126.25, 60.11, 54.85, 54.82, 52.63, 45.72, 33.13, 29.66, 20.14, 17.15, 13.97; LCMS C25H36N4S method (B) Rt=4.593 min, ESI+ m/z=425.3 (M+H).
1-iso-Butyl-1-(2-methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethoxy)pyridin-3-yl)phenyl)thiourea OR0608-2 (90%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 8.31 (d, J=2.5 Hz, 1H), 7.73 (dd, J=8.6, 2.5 Hz, 1H), 7.47 (dd, J=8.0, 1.9 Hz, 1H), 7.41 (d, J=8.0 Hz, 1H), 7.31 (d, J=1.9 Hz, 1H), 6.84 (d, J=8.6 Hz, 1H), 5.49 (brs, 2H), 4.50 (t, J=5.7 Hz, 2H), 3.39-3.31 (m, 2H), 2.92-2.84 (m, 2H), 2.81 (brs, 8H), 2.48 (s, 3H), 2.27 (s, 3H), 1.99-1.98 (m, 1H), 0.97 (d, J=6.8 Hz, 6H); LCMS C24H35N5OS method (B) Rt=4.530 min, ESI+ m/z=442.3 (M+H).
N-((1-(3-(1-iso-Butylthioureido)-4-methylphenyl)-1H-1,2,3-triazol-4-yl)methyl)methanesulfonamide OR0609-2 (95%) as a white solid. 1H NMR (400 MHz, DMSO) δ 8.75 (s, 1H), 7.82-7.52 (m, 4H), 4.31 (s, 2H), 3.60 (brs, 2H), 2.95 (s, 3H), 2.18 (s, 3H), 1.90-1.78 (m, 1H), 0.96-0.86 (m, 6H); LCMS C16H24N6O2S2 method (B) Rt=5.127 min, ESI+ m/z=397.2 (M+H).
tert-Butyl (2-(1-(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)thioureido)ethyl)carbamate OR0610-3 (98%) as a pale yellow solid, engaged in the next step without further purification. 1H NMR (400 MHz, CDCl3) δ 7.53 (dd, J=7.9, 1.4 Hz, 1H), 7.49 (d, J=8.1 Hz, 2H), 7.40-7.36 (m, 2H), 7.26 (d, J=8.1 Hz, 2H), 5.46 (brs, 1H), 4.80-4.72 (m, 1H), 3.86-3.78 (m, 1H), 3.54-3.38 (m, 2H), 2.89-2.81 (m, 2H), 2.72-2.64 (m, 2H), 2.63 (brs, 8H), 2.38 (s, 3H), 2.25 (s, 3H), 1.39 (s, 9H); LCMS C28H41N5O2S method (B) Rt=4.513 min, ESI+ m/z=512.3 (M+H).
tert-Butyl (3-(1-(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)thioureido)propyl)carbamate OR0611-3 (90%) a pale yellow solid, engaged in the next step without further purification. 1H NMR (400 MHz, CDCl3) δ 7.53 (dd, J=7.9, 1.8 Hz, 1H), 7.46 (d, J=8.1 Hz, 2H), 7.39 (d, J=7.9 Hz, 1H), 7.31 (d, J=1.8 Hz, 1H), 7.28 (d, J=8.2 Hz, 1H), 5.56 (brs, 2H), 5.38 (brs, 1H), 4.74-4.66 (m, 1H), 3.72-3.63 (m, 1H), 3.35-3.13 (m, 2H), 2.88-2.80 (m, 2H), 2.67-2.59 (m, 2H), 2.52 (brs, 8H), 2.31 (s, 3H), 2.25 (s, 3H), 1.94-1.71 (m, 2H), 1.42 (s, 9H); LCMS C29H43N5O2S method (B) Rt=4.607 min, ESI+ m/z=526.3 (M+H).
1-(2-Methyl-5-(6-(4-methylpiperazine-1-carbonyl)pyridin-3-yl)phenyl)-1-propylthiourea OR0612-2 (82%) as a yellow solid. LCMS C22H29N5OS method (B) Rt=4.215 min, ESI+ m/z=412.2 (M+H).
4-Methyl-N-(5-(4-methyl-3-(1-propylthioureido)phenyl)pyridin-2-yl)piperazine-1-carboxamide OR0616-2 (88%) as a yellow solid. LCMS C22H30N6OS method (B) Rt=4.309 min, ESI+ m/z=427.3 (M+H).
1-(5-Fluoro-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)-1-propylthiourea OR0633-2 (97%) as a yellow solid. LCMS C22H29FN4O2S2 method (B) Rt=4.870 min, ESI+ m/z=465.2 (M+H).
General Procedure for the Synthesis of N-([1,1′-biaryl]-3-ylcarbamothioyl)benzamide Derivatives. As previously described above.
N-((4-Methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)(propyl)carbamothioyl)benzamide OR0600-3 (quantitative), as a pale yellow solid, was used in the next step without further purification. LCMS C31H38N4OS method (B) Rt=4.868 min, ESI+ m/z=515.3 (M+H).
N-(iso-Butyl(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)carbamothioyl)benzamide OR0603-3 (quantitative) as a white solid. LCMS C32H40N4OS method (B) Rt=5.080 min, ESI+ m/z=529.6 (M+H).
N-((Cyclopropylmethyl)(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)carbamothioyl)benzamide OR0604-3 (quantitative) as a white solid. LCMS C32H3SN4OS method (B) Rt=4.915 min, ESI+ m/z=527.3 (M+H).
N-(iso-Pentyl(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)carbamothioyl)benzamide OR0605-3 (96%), as a light yellow solid, used in the next step without further purification. Rf=0.40 (DCM-MeOH—NH4OH, 90:9:1); LCMS C33H42N4OS method (B) Rt=5.154 min, ESI+ m/z=543.3 (M+H).
N-(Butyl(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)carbamothioyl)benzamide OR0606-3 (96%) as a light yellow solid, used in the next step without further purification. Rf=0.40 (DCM-MeOH—NH4OH, 90:9:1); LCMS C32H40N4OS method (B) Rt=5.059 min, ESI+ m/z=529.3 (M+H).
N-(iso-Butyl(2-methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethoxy)pyridin-3-yl)phenyl)carbamothioyl)benzamide OR0608-3 (quantitative) as a white solid. LCMS C31H39N5O2S method (B) Rt=5.074 min, ESI+ m/z=546.3 (M+H).
N-(iso-Butyl(2-methyl-5-(4-(methylsulfonamidomethyl)-1H-1,2,3-triazol-1-yl)phenyl)carbamothioyl)benzamide OR0609-3 (quantitative) as a white solid. LCMS C23H28N6O3S2 method (B) Rt=5.882 min, ESI+ m/z=501.2 (M+H).
tert-Butyl (2-(3-benzoyl-1-(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)thioureido)ethyl) carbamate OR0610-4 (96%) as a pale yellow solid, used in the next step without further purification. LCMS C35H45N5O3S method (B) Rt=4.938 min, ESI+ m/z=616.3 (M+H).
tert-Butyl (3-(3-benzoyl-1-(4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)thioureido)propyl) carbamate OR0611-4 (98%) as a light yellow solid, used in the next step without further purification. LCMS C36H47N5O3S method (B) Rt=4.936 min, ESI+ m/z=630.3 (M+H).
N-((2-Methyl-5-(6-(4-methylpiperazine-1-carbonyl)pyridin-3-yl)phenyl)(propyl)carbamothioyl)benzamide OR0612-3 (quantitative) as a beige solid. LCMS C29H33N5O2S method (B) Rt=4.773 min, ESI+ m/z=516.3 (M+H).
N-(5-(3-(3-Benzoyl-1-propylthioureido)-4-methylphenyl)pyridin-2-yl)-4-methylpiperazine-1-carboxamide OR0616-3 (quantitative) as a yellow solid. LCMS C29H34N6O2S method (B) Rt=4.875 min, ESI+ m/z=531.3 (M+H).
N-((5-Fluoro-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-yl)(propyl)carbamothioyl)benzamide OR0633-3 (quantitative) as a yellow solid. LCMS C29H33FN403S2 method (B) Rt=5.434 min, ESI+ m/z=569.2 (M+H).
General Procedure for the Synthesis of [1,1′-biaryl]-3-N-alkylamine derivatives. As previously described above.
N-iso-Butyl-4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-amine OR0603-4 (75%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 7.50 (d, J=8.2 Hz, 2H), 7.25 (d, J=8.2 Hz, 2H), 7.10 (d, J=7.7 Hz, 1H), 6.84 (dd, J=7.7, 1.6 Hz, 1H), 6.77 (d, J=1.6 Hz, 1H), 3.60 (brs, 1H), 3.04 (d, J=6.7 Hz, 2H), 2.88-2.80 (m, 2H), 2.69-2.61 (m, 2H), 2.53 (brs, 8H), 2.32 (s, 3H), 2.17 (s, 3H), 2.02-1.92 (m, 1H), 1.02 (d, J=6.7 Hz, 6H); LCMS C24H35N3 method (B) Rt=4.324 min, ESI+ m/z=366.4 (M+H).
N-(Cyclopropylmethyl)-4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-amine OR0604-4 (71%) as a colorless oil. 1H NMR (400 MHz, CDCl3) δ 7.50 (d, J=8.1 Hz, 2H), 7.25 (d, J=8.1 Hz, 2H), 7.10 (d, J=7.8 Hz, 1H), 6.86 (dd, J=7.8, 1.6 Hz, 1H), 6.77 (d, J=1.6 Hz, 1H), 3.06 (d, J=6.9 Hz, 2H), 2.87-2.81 (m, 2H), 2.68-2.60 (m, 2H), 2.51 (brs, 8H), 2.31 (s, 3H), 2.20 (s, 3H), 1.20-1.14 (m, 1H), 0.61-0.55 (m, 2H), 0.30-0.24 (m, 2H); LCMS C24H33N3 method (B) Rt=4.392 min, ESI+ m/z=364.3 (M+H).
N-iso-Pentyl-4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-amine OR0605-4 (68%) as a light yellow oil. Rf=0.45 (DCM-MeOH—NH4OH, 90:9:1); LCMS C25H37N3 method (B) Rt=4.966 min, ESI+ m/z=380.6 (M+H).
N-Butyl-4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-amine OR0606-4 (63%) as a colorless oil, used in the next step without further purification. Rf=0.45 (DCM-MeOH—NH4OH, 90:9:1); LCMS C24H35N3 method (B) Rt=4.590 min, ESI+ m/z=366.3 (M+H).
N-iso-Butyl-2-methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethoxy)pyridin-3-yl)aniline OR0608-4 (44%) as a colorless oil. LCMS C23H34N4O method (B) Rt=4.859 min, ESI+ m/z=383.3 (M+H).
N-((1-(3-(iso-Butylamino)-4-methylphenyl)-1H-1,2,3-triazol-4-yl)methyl)methanesulfonamide OR0609-4 (68%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 8.02 (s, 1H), 7.32-7.28 (m, 1H), 7.20 (d, J=8.0 Hz, 1H), 7.05-7.00 (m, 1H), 5.44 (brs, 1H), 4.55 (d, J=5.4 Hz, 2H), 3.08 (d, J=6.8 Hz, 2H), 3.03 (s, 3H), 2.31 (s, 3H), 2.14-2.04 (m, 1H), 1.09 (d, J=6.8 Hz, 6H); LCMS C15H23N5O2S method (B) Rt=5.794 min, ESI+m/z=338.2 (M+H).
tert-Butyl (2-((4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)ethyl)carbamate OR0610-5 (35%) as a colorless oil. Rf=0.65 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.49 (d, J=8.1 Hz, 2H), 7.24 (d, J=8.1 Hz, 2H), 7.10 (d, J=7.6 Hz, 1H), 6.85 (dd, J=7.6, 1.2 Hz, 1H), 6.75 d, J=1.2 Hz, 1H), 4.88 (brs, 1H), 4.18 (brs, 1H), 3.51-3.43 (m, 2H), 3.37-3.29 (m, 2H), 2.89-2.81 (m, 2H), 2.70-2.62 (m, 2H), 2.57 (brs, 8H), 2.35 (s, 3H), 2.17 (s, 3H), 1.45 (s, 9H); 13C NMR (100 MHz, CDCl3) δ 156.90, 146.39, 140.17, 139.99, 138.80, 130.42, 128.94, 127.16, 121.22, 115.69, 107.98, 79.68, 60.35, 54.96, 52.87, 45.86, 45.20, 40.01, 33.14, 28.39, 17.20; LCMS C27H40N4O2 method (B) R=4.851 min, ESI+ m/z=453.4 (M+H).
tert-Butyl (3-((4-methyl-4′-(2-(4-methylpiperazin-1-yl)ethyl)-[1,1′-biphenyl]-3-yl)amino)propyl)carbamate OR0611-5 (46%) as a colorless oil. Rf=0.65 (DCM-MeOH—NH4OH, 90:9:1); 1H NMR (400 MHz, CDCl3) δ 7.49 (d, J=8.2 Hz, 2H), 7.24 (d, J=8.2 Hz, 2H), 7.10 (d, J=7.8 Hz, 1H), 6.85 (dd, J=7.8, 1.6 Hz, 1H), 6.78 (d, J=1.6 Hz, 1H), 4.70 (brs, 1H), 3.32-3.24 (m, 4H), 2.89-2.81 (m, 2H), 2.71-2.63 (m, 2H), 2.61 (brs, 8H), 2.36 (s, 3H), 2.18 (s, 3H), 1.84 (quint, J=6.6 Hz, 2H), 1.44 (s, 9H); LCMS C28H42N4O2 method (B) R=4.818 min, ESI+ m/z=467.4 (M+H).
(5-(4-Methyl-3-(propylamino)phenyl)pyridin-2-yl)(4-methylpiperazin-1-yl)methanone OR0612-4 (95%) as a beige oil. 1H NMR (400 MHz, CDCl3) δ 8.79 (d, J=2.1 Hz, 1H), 7.97 (dd, J=8.1, 2.1 Hz, 1H), 7.71 (d, J=8.1 Hz, 1H), 7.15 (d, J=7.8 Hz, 1H), 6.85 (dd, J=7.5, 1.8 Hz, 1H), 6.77 (d, J=1.8 Hz, 1H), 3.89 (brs, 2H), 3.76 (brs, 2H), 3.20 (t, J=7.1 Hz, 2H), 2.59 (brs, 2H), 2.51 (brs, 2H), 2.37 (s, 3H), 2.19 (s, 3H), 1.73 (sext, J=7.4 Hz, 2H), 1.05 (t, J=7.4 Hz, 3H); LCMS C21H28N4O method (B) Rt=4.358 min, ESI+ m/z=353.3 (M+H).
4-Methyl-N-(5-(4-methyl-3-(propylamino)phenyl)pyridin-2-yl)piperazine-1-carboxamide OR0616-4 (90%) as a beige oil. 1H NMR (400 MHz, CDCl3) δ 8.40 (d, J=2.2 Hz, 1H), 8.05 (d, J=8.7 Hz, 1H), 7.87 (dd, J=8.7, 2.2 Hz, 1H), 7.38 (brs, 1H), 7.11 (d, J=7.7 Hz, 1H), 6.81 (dd, J=7.7, 1.7 Hz, 1H), 6.73 (d, J=1.7 Hz, 1H), 3.65-3.59 (m, 4H), 3.19 (t, J=7.1 Hz, 2H), 2.57-2.53 (m, 4H), 2.39 (s, 3H), 2.17 (s, 3H), 1.72 (sext, J=7.4 Hz, 2H), 1.04 (t, J=7.4 Hz, 3H); LCMS C21H29N5O method (B) Rt=4.217 min, ESI+ m/z=368.3 (M+H).
5-Fluoro-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-N-propyl-[1,1′-biphenyl]-3-amine OR0633-4 (55%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 7.78 (d, J=8.5 Hz, 2H), 7.69 (d, J=8.5 Hz, 2H), 6.65 (dd, J=10.2, 1.6 Hz, 1H), 6.55 (d, J=1.6 Hz, 1H), 3.20 (t, J=7.4 Hz, 2H), 3.19 (brs, 4H), 2.65 (brs, 4H), 2.38 (s, 3H), 2.08 (s, 3H), 1.72 (sext, J=7.4 Hz, 2H), 1.05 (t, J=7.4 Hz, 3H); LCMS C21H28FN3O2S method (B) Rt=5.345 min, ESI+ m/z=406.2 (M+H).
General Procedure for the Synthesis of [1,1′-biaryl]-3-amine Derivatives. Method (A): a suspension of appropriate 3-nitro-1,1′-biaryl (1 mmol), iron powder (280 mg, 5 mmol) and ammonium chloride (539 mg, 10 mmol) in a mixture of water-ethanol (1:1, 20 mL) was refluxed until the reaction was complete as indicated by TLC monitoring. The reaction mixture was filtrated through a short pad of celite and rinsed with EtOH. The solvent was distillated off under reduced pressure and the residue was purified by flash chromatography to afford the corresponding [1,1′-biaryl]-3-amine. Method (B): as already described for 3-nitro-1,1′-biaryl derivatives, starting from commercially available 3-amino-4-methylphenylboronic acid and appropriate aryl halide.
2-Methyl-5-(6-(2-(4-methylpiperazin-1-yl)ethoxy)pyridin-3-yl)aniline OR0608-5 (method B, 85%) as a colorless oil. 1H NMR (400 MHz, CDCl3) δ 8.31 (d, J=2.5 Hz, 1H), 7.74 (dd, J=8.6, 2.5 Hz, 1H), 7.11 (d, J=7.7 Hz, 1H), 6.85 (dd, J=7.7, 1.8 Hz, 1H), 6.82 (d, J=1.8 Hz, 1H), 6.79 (d, J=8.6 Hz, 1H), 4.48 (t, J=5.8 Hz, 2H), 3.70 (brs, 2H), 2.84 (t, J=5.8 Hz, 2H), 2.69 (brs, 4H), 2.58 (brs, 4H), 2.35 (s, 3H), 2.20 (s, 3H); LCMS C19H26N4O method (B) Rt=3.514 min, ESI+ m/z=327.2 (M+H).
N-((1-(3-Amino-4-methylphenyl)-1H-1,2,3-triazol-4-yl)methyl)methanesulfonamide OR0609-5 (method A, 97%) as a pale yellow solid. LCMS C19H15N5O2S method (B) R=4.179 min, ESI+ m/z=282.2 (M+H).
(5-(3-Amino-4-methylphenyl)pyridin-2-yl)(4-methylpiperazin-1-yl)methanone OR0612-5 (method B, 90%) as a beige solid. 1H NMR (400 MHz, CDCl3) δ 8.75 (s, 1H), 7.92 (dd, J=8.1, 1.9 Hz, 1H), 7.69 (d, J=8.1 Hz, 1H), 7.15 (d, J=7.7 Hz, 1H), 6.91 (d, J=7.7 Hz, 1H), 6.88 (s, 1H), 3.87 (brs, 2H), 3.76 (brs, 2H), 3.72 (brs, 2H), 2.56 (brs, 2H), 2.47 (brs, 2H), 2.35 (s, 3H), 2.21 (s, 3H); LCMS C15H22N4O method (B) Rt=2.725 min, ESI+ m/z=311.2 (M+H).
N-(5-(3-Amino-4-methylphenyl)pyridin-2-yl)-4-methylpiperazine-1-carboxamide OR0616-5 (method B, 70%) as an orange solid. 1H NMR (400 MHz, CDCl3) δ 8.37 (d, J=2.2 Hz, 1H), 8.04 (d, J=8.7 Hz, 1H), 7.83 (dd, J=8.7, 2.2 Hz, 1H), 7.43 (brs, 1H), 7.11 (d, J=7.7 Hz, 1H), 6.88 (dd, J=7.7, 1.7 Hz, 1H), 6.85 (d, J=1.7 Hz, 1H), 3.62-3.57 (m, 4H), 2.53-2.47 (m, 4H), 2.36 (s, 3H), 2.20 (s, 3H); LCMS C15H23N5O method (B) Rt=2.298 min, ESI+m/z=326.1 (M+H).
5-Fluoro-4-methyl-4′-((4-methylpiperazin-1-yl)sulfonyl)-[1,1′-biphenyl]-3-amine OR0633-5 (method A, quantitative) as a yellow oil. LCMS C18H22FN3O2S method (B) Rt=4.657 min, ESI+ m/z=364.1 (M+H).
1-(2-((5-Bromopyridin-2-yl)oxy)ethyl)-4-methylpiperazine OR0608-6′ From commercially available 5-bromo-2-fluoropyridine and 2-(4-methylpiperazin-1-yl)ethan-1-ol, (62%) as a colorless oil. 1H NMR (400 MHz, CDCl3) δ 8.16 (d, J=2.5 Hz, 1H), 7.62 (dd, J=8.8, 2.5 Hz, 1H), 6.66 (d, J=8.8 Hz, 1H), 4.40 (t, J=5.8 Hz, 2H), 2.78 (t, J=5.8 Hz, 2H), 2.63 (brs, 4H), 2.52 (brs, 4H), 2.31 (s, 3H); LCMS C12H18BrN3O method (B) Rt=3.571 min, ESI+ m/z=300.0 (M+H).
N-((1-(4-Methyl-3-nitrophenyl)-1H-1,2,3-triazol-4-yl)methyl)methanesulfonamide OR0609-6
To a mixture of N-(prop-2-yn-1-yl)methanesulfonamide (620 mg, 4.66 mmol), CuSO4 pentahydrate (30 mg, 0.12 mmol), sodium ascorbate (118 mg, 0.60 mmol) in a mixture of tert-butanol-H2O (1:1, 30 mL) was added 4-azido-1-methyl-2-nitrobenzeneii (817 mg, 4.59 mmol).
The suspension was stirred for 48 hrs at room temperature, until consumption of starting material monitored by LCMS. The solvent was distillated off under reduced pressure and the residue was taken up into 10% aqueous NH4OH solution (20 mL) and extracted with EtOAc (2×50 mL). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The residue was purified by flash chromatography, gradient DCM-EtOAc (100:0 to 65:35) to afford N-((1-(4-Methyl-3-nitrophenyl)-1H-1,2,3-triazol-4-yl)methyl)methanesulfonamide OR0609-6 (1.24 g, 87%) as a beige solid. 1H NMR (400 MHz, DMSO) δ 8.87 (s, 1H), 8.53 (d, J=2.3 Hz, 1H), 8.20 (dd, J=8.3, 2.3 Hz, 1H), 7.74 (d, J=8.3 Hz, 1H), 7.64 (s, 1H), 4.33 (s, 2H), 2.96 (s, 3H), 2.57 (s, 3H); LCMS C11H13N5O4S method (B) R=5.027 min, ESI+ m/z=312.1 (M+H).
N-(5-Bromopyridin-2-yl)-4-methylpiperazine-1-carboxamide OR0616-6
From 4-nitrophenyl (5-bromopyridin-2-yl)carbamate OR0616-7, (79%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 8.23 (d, J=2.4 Hz, 1H), 7.96 (d, J=8.9 Hz, 1H), 7.73 (dd, J=8.9, 2.4 Hz, 1H), 3.61-3.54 (m, 4H), 2.54-2.47 (m, 4H), 2.36 (s, 3H).
4-Nitrophenyl (5-bromopyridin-2-yl)carbamate OR0616-7iii
From commercially available 5-bromopyridin-2-amine and 4-nitrophenyl chloroformate, (43%) as a white powder. LCMS C12H8BrN3O4 method (B) Rt=6.490 min, ESI+ m/z=338.0 (M+H).
1-((3′-Fluoro-4′-methyl-5′-nitro-[1,1′-biphenyl]-4-yl)sulfonyl)-4-methylpiperazine OR0633-6
As previously described for 3-nitro-1,1′-biaryl derivatives, from commercially available 5-bromo-1-fluoro-2-methyl-3-nitrobenzene and 1-methyl-4-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)sulfonyl)piperazine, (quantitative) as a yellow oil. LCMS C18H20FN3O4S method (B) Rt=5.007 min, ESI+ m/z=394.2 (M+H).
General Procedure for the Synthesis of 4,4,5,5-tetramethyl-2-aryl-1,3,2-dioxaborolane. Method (A); under argon, a suspension of appropriate aryl halide (5.6 mmol), bis(pinacolato)diboron (3.63 g, 14.3 mmol), potassium acetate (2.80 g, 28.6 mmol), PdCl2(dppf) (697 mg, 0.9 mmol), in degassed 1,4-dioxane (50 mL) was refluxed for 2 hrs upon complete consumption of starting material. The resulting mixture was successively allowed to cool to room temperature, filtered through a short pad of celite and washed with EtOAc. The solvent was distillated off under reduced pressure and the residue was purified by flash chromatography, to afford corresponding 4,4,5,5-tetramethyl-2-aryl-1,3,2-dioxaborolane. Method (B); under argon, at −78° C., to a solution of appropriate aryl halide (1.0 mmol) in tetrahydrofuran (5 mL) were added successively 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.37 mL, 1.8 mmol) and then n-BuLi (2.5 M in hexane, 0.47 mL, 1.18 mmol) over 5 minutes. After addition, the resulting mixture was stirred at −78° C. for 30 min and then quenched with water (5 mL). The aqueous layer was successively washed with DCM (3×5 mL), acidified with 2M aqueous HCl solution until pH=7, and extracted with EtOAc (3×5 mL). The combined organic layers were dried on Na2SO4 and the solvent was distillated off under reduced pressure affording expected 4,4,5,5-tetramethyl-2-aryl-1,3,2-dioxaborolane.
Ethyl 2-((2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)(propyl)amino)thiazole-4-carboxylate OR0625-2 (method A, quantitative) as a white solid. 1H NMR (400 MHz, CDCl3) δ 7.73 (dd, J=7.5, 1.2 Hz, 1H), 7.62 (d, J=1.2 Hz, 1H), 7.35 (d, J=7.5 Hz, 1H), 7.26 (s, 1H), 4.35 (q, J=7.1 Hz, 2H), 4.02-3.75 (m, 2H), 2.24 (s, 3H), 1.67 (sext, J=7.4 Hz, 2H), 1.37 (t, J=7.1 Hz, 3H), 1.34 (s, 12H), 0.92 (t, J=7.4 Hz, 3H); LCMS C22H31BN2O4S method (B) Rt=6.109 min, ESI+ m/z=349.2 (ArB(OH)2+H).
1-Methyl-4-((5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)sulfonyl)piperazine OR0627-2 (method B, 43%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 9.00 (d, J=1.7 Hz, 1H), 8.26 (dd, J=7.7, 1.7 Hz, 1H), 7.88 (d, J=7.7 Hz, 1H), 3.50 (brs, 4H), 2.71 (brs, 4H), 2.45 (s, 3H), 1.36 (s, 12H); LCMS C16H26BN3O4S method (B) Rt=1.149 min. ESI+ m/z=286.1 (ArB(OH)2+H).
1-Methyl-4-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-(trifluoromethyl)phenyl)sulfonyl)piperazine OR0642-2 (method A, 83%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 8.01 (s, 1H), 7.92 (d, J=7.8 Hz, 1H), 7.87 (dd, J=7.8, 1.4 Hz, 1H), 3.17 (brs, 4H), 2.63 (brs, 4H), 2.38 (s, 3H), 1.38 (s, 12H); LCMS C18H26BF3N2O4S method (B) Rt=4.951 min, ESI+ m/z=435.2 (M+H).
(4-Methyl-6-((4-methylpiperazin-1-yl)sulfonyl)pyridin-3-yl)boronic acid OR0652-2 (method B), aqueous layer was acidified until pH=3. The aqueous layer was washed with DCM (3×50 mL) and then concentrated under reduced pressure to afford (4-methyl-6-((4-methylpiperazin-1-yl)sulfonyl)pyridin-3-yl)boronic acid OR0652-2 (40%) as a white solid. LCMS C11H18BN3O4S method (B) Rt=1.542 min, ESI+ m/z=300.2 (M+H).
General Procedure for the Synthesis of 1-(4-halogeno-benzenesulfonyl)-4-methyl-piperazine Derivative. Under argon, at 0° C., to a solution of appropriate 4-halogenobenzenesulfonyl chloride (1.0 mmol) in dichloromethane (6 mL) was added 1-methyl-piperazine (125 μL, 1.12 mmol). The reaction mixture was stirred at 0° C. for 30 min. and allowed to warm to room temperature. After stirring for 2 hours, the mixture was diluted with DCM (15 mL), then saturated Na2CO3 aqueous solution was added (5 mL) and the aqueous layer was extracted with DCM (2×15 mL). The combined organic layers were dried over Na2SO4 and the solvent was distillated off under reduced pressure to afford corresponding 1-(4-halogeno-benzenesulfonyl)-4-methyl-piperazine used in the next step without further purification.
1-((5-Bromopyridin-2-yl)sulfonyl)-4-methylpiperazine OR0627-3 (quantitative). 1H NMR (400 MHz, CDCl3) δ 8.73 (d, J=2.0 Hz, 1H), 8.03 (dd, J=8.3, 2.0 Hz, 1H), 7.81 (d, J=8.3 Hz, 1H), 3.38-3.30 (m, 4H), 2.52-2.44 (m, 4H), 2.29 (s, 3H); LCMS C10H14BrN3O2S method (B) Rt=3.703 min, ESI+ m/z=320.0 (M+H).
1-((4-Bromo-3-(trifluoromethyl)phenyl)sulfonyl)-4-methylpiperazine OR0642-3 (99%). 1H NMR (400 MHz, CDCl3) δ 8.02 (d, J=1.9 Hz, 1H), 7.90 (d, J=8.3 Hz, 1H), 7.74 (dd, J=8.3, 2.0 Hz, 1H), 3.12-3.04 (m, 4H), 2.56-2.48 (m, 4H), 2.29 (s, 3H); LCMS C10H14BrN3O2S method (B) Rt=4.419 min. ESI+ m/z=387.0 (M+H).
1-((5-Bromo-4-methylpyridin-2-yl)sulfonyl)-4-methylpiperazine OR0652-3 (63%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 8.69 (s, 1H), 7.78 (s, 1H), 3.41-3.32 (m, 4H), 2.56-2.50 (m, 4H), 2.49 (s, 3H), 2.32 (s, 3H); LCMS C11H16BrN3O2S method (B) Rt=4.075 min, ESI+ m/z=334.0 (M+H).
N-(5-Bromopyridin-2-yl)-4-methylpiperazine-1-sulfonamide OR0625-4
Using a modified procedure described by Borghese et al.,iv a mixture of N-(5-bromopyridin-2-yl)-2-oxooxazolidine-3-sulfonamide OR0625-5 (1.73 g, 5.37 mmol), 1-methylpiperazine (600 μL, 5.37 mmol) and triethylamine (2.25 mL, 16.13 mmol) in acetonitrile (27 mL) was refluxed for 4 days. Upon complete consumption of the starting material monitored by LCMS, the solvent was distillated off under reduced pressure, then saturated aqueous NaHCO3 solution was added until pH=8, then the aqueous layer was extracted with DCM (3×40 mL). The combined organic layers were dried over Na2SO4, and the solvent was distillated off under reduced pressure. The residue was purified by flash chromatography, gradient DCM-MeOH—NH4OH (100:0:0 to 90:9:1) to afford N-(5-bromopyridin-2-yl)-4-methylpiperazine-1-sulfonamide OR0625-4 (311 mg, 17%) as a white solid. 1H NMR (400 MHz, CDCl3) δ 8.44 (d, J=2.4 Hz, 1H), 7.78 (dd, J=8.9, 2.4 Hz, 1H), 7.28 (d, J=8.9 Hz, 1H), 3.39-3.32 (m, 4H), 2.55-2.48 (m, 4H), 2.33 (s, 3H); LCMS C10H15BrN4O2S method (B) Rt=3.567 min, ESI+ m/z=335.0 (M+H).
N-(5-Bromopyridin-2-yl)-2-oxooxazolidine-3-sulfonamide OR0625-5
Using a modified procedure described by Borghese et al.,iv under argon, at 0° C., to a solution of chlorosulfonyl isocyanate (503 μL, 5.78 mmol) in dry dichloromethane (20 mL) was added dropwise 2-chloroethan-1-ol (390 μL, 5.78 mmol). After stirring at 0° C. for 1 hour, triethylamine (2.5 mL, 17.30 mmol) was added, followed by a solution of 5-bromopyridin-2-amine (1.0 g, 5.78 mmol) in dry dichloromethane (15 mL) dropwise. The resulting mixture was stirred for an additional hour at 0° C., then allowed to warm to room temperature and stirred overnight. The reaction was quenched by addition of 0.25N HCl aqueous solution, extracted with DCM (2×40 mL), and the combined organic layers were dried over anhydrous Na2SO4. The solvent was distillated off under reduced pressure and the crude residue was triturated with cyclohexane affording N-(5-bromopyridin-2-yl)-2-oxooxazolidine-3-sulfonamide OR0625-5 (1.73 g, 93%) as a white solid. LCMS C8H8BrN3O4S method (B) Rt=4.761 min, ESI+ m/z=322.0 (M+H).
2-((5-Bromopyridin-2-yl)oxy)-1-(4-methylpiperazin-1-yl)ethan-1-one OR0630-2 By peptide coupling from commercially available 2-((5-bromopyridin-2-yl)oxy)acetic acid, using same procedure as described for di-tert-butyl (2-(2-((3-benzamidoaryl)amino)thiazol-4-yl)pyrimidin-4-yl)carbamate derivatives. 1H NMR (400 MHz, CDCl3) δ 8.13 (d, J=2.4 Hz, 1H), 7.67 (dd, J=8.8, 2.4 Hz, 1H), 6.80 (d, J=8.8 Hz, 1H), 4.97 (s, 2H), 3.67 (brs, 2H), 3.53 (brs, 2H), 2.48 (brs, 2H), 2.45 (brs, 2H), 2.34 (s, 3H); LCMS C12H16BrN3O2 method (B) Rt=3.649 min, ESI+ m/z=314.0 (M+H).
General Procedure for the Synthesis of ethyl 2-((5-halogeno-2-methylphenyl)amino)thiazole-4-carboxylate Derivative and analogues. Method (A): as previously described for synthesis of ethyl 2-([1,1′-biaryl]-3-ylamino)thiazole-4-carboxylate derivatives, method (A). Method (B): To a suspension of ethyl 2-((5-halogeno-2-methylphenyl)amino)thiazole-4-carboxylate or analogue (0.16 mol) and cesium carbonate (73 g, 0.22 mol) in dimethylformamide (1 L) was added appropriate alkyl halide (20 mL, 0.22 mol). The reaction mixture was stirred at r.t for 72 hrs until complete consumption of starting material. The solvent was distillated off under reduced pressure and the residue was poured onto water (300 mL) and extracted with Et2O (3×300 mL). The combined organic layers were dried over Na2SO4 and concentrated under vacuum. The residue was purified by flash chromatography, to afford corresponding ethyl 2-((5-halogeno-2-methylphenyl)(alkyl)amino)thiazole-4-carboxylate or analogue.
Ethyl 2-((5-iodo-2-methylphenyl)(propyl)amino)thiazole-4-carboxylate OR0607-2 (method A, 73%) as a pale yellow solid. Rf=0.20 (DCM-PE, 80:20); 1H NMR (400 MHz, CDCl3) δ 7.62 (dd, J=8.1, 1.8 Hz, 1H), 7.54 (d, J=1.8 Hz, 1H), 7.30 (s, 1H), 7.08 (d, J=8.1 Hz, 1H), 4.35 (q, J=7.1 Hz, 2H), 3.88 (brs, 2H), 2.17 (s, 3H), 1.64 (sext, J=7.4 Hz, 2H), 1.37 (t, J=7.1 Hz, 3H), 0.93 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.27, 161.88, 144.08, 143.88, 138.22, 137.96, 137.26, 133.88, 116.96, 91.05, 61.20, 53.96, 21.11, 17.42, 14.47, 11.37; LCMS C16H19IN2O2S method (B) Rt=8.676 min, ESI+ m/z=431.0 (M+H).
Ethyl 2-((5-bromo-2-methylphenyl)(propyl)amino)thiazole-4-carboxylate OR0625-3 (method B, 54%) as a light brown solid. Rf=0.31 (DCM); 1H NMR (400 MHz, CDCl3) δ 7.44 (dd, J=8.2, 2.0 Hz, 1H), 7.36 (d, J=2.0 Hz, 1H), 7.31 (s, 1H), 7.22 (d, J=8.2 Hz, 1H), 4.35 (q, J=7.1 Hz, 2H), 3.89 (brs, 2H), 2.17 (s, 3H), 1.64 (sext, J=7.4 Hz, 2H), 1.37 (t, J=7.1 Hz, 3H), 0.94 (t, J=7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.27, 161.90, 143.95, 143.90, 136.50, 133.63, 132.39, 132.06, 120.22, 117.00, 61.22, 53.95, 21.13, 17.34, 14.47, 11.37; LCMS C16H19BrN2O2S method (B) Rt=7.376 min, ESI+ m/z=383.0 (M+H).
Ethyl 2-((5-bromo-2-methylphenyl)(methyl)amino)thiazole-4-carboxylate OR0631-2 (method B, 37%) as a light brown solid. Rf=0.29 (DCM); 1H NMR (400 MHz, CDCl3) δ 7.43 (dd, J=8.2, 2.0 Hz, 1H), 7.39 (d, J=2.0 Hz, 1H), 7.35 (s, 1H), 7.22 (d, J=8.2 Hz, 1H), 4.36 (q, J=7.1 Hz, 2H), 3.50 (s, 3H), 2.18 (s, 3H), 1.38 (t, J=7.1 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.17, 161.80, 145.38, 143.87, 135.77, 133.61, 132.13, 131.39, 120.39, 117.47, 61.34, 40.01, 27.04, 17.17, 14.48; LCMS C14H15BrN2O2S method (B) Rt=7.212 min, ESI+m/z=355.0 (M+H).
Ethyl 2-((5-bromo-2-methylphenyl)(ethyl)amino)thiazole-4-carboxylate OR0632-2 (method B, 41%) as a light brown solid. Rf=0.29 (DCM); 1H NMR (400 MHz, CDCl3) δ 7.44 (dd, J=8.2, 2.0 Hz, 1H), 7.36 (d, J=2.0 Hz, 1H), 7.31 (s, 1H), 7.23 (d, J=8.2 Hz, 1H), 4.35 (q, J=7.1 Hz, 2H), 4.01 (brs, 2H), 2.18 (s, 3H), 1.37 (t, J=7.1 Hz, 3H), 1.21 (t, J=7.2 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 170.00, 161.96, 144.01, 143.57, 136.65, 133.57, 132.52, 132.09, 120.23, 117.05, 61.21, 46.91, 17.33, 14.49, 13.12; LCMS C15H17BrN2O2S method (B) R=7.570 min, ESI+ m/z=369.0 (M+H).
Ethyl 2-((5-bromo-2-methylphenyl)(isopropyl)amino)thiazole-4-carboxylate OR0636-2 (method B, 67%) as a light yellow solid. Rf=0.34 (DCM); 1H NMR (400 MHz, CDCl3) δ 7.47 (dd, J=8.2, 2.1 Hz, 1H), 7.33 (d, J=2.1 Hz, 1H), 7.28 (s, 1H), 7.24 (d, J=8.2 Hz, 1H), 5.07 (sept, J=6.7 Hz, 1H), 4.35 (q, J=7.1 Hz, 2H), 2.18 (s, 3H), 1.37 (t, J=7.1 Hz, 3H), 1.25-1.23 (m, 6H); 13C NMR (100 MHz, CDCl3) δ 169.99, 161.99, 143.95, 141.33, 138.00, 133.75, 133.56, 132.32, 119.97, 116.64, 61.14, 52.25, 20.76, 17.83, 14.49; LCMS C16H19BrN2O2S method (B) Rt=7.818 min, ESI+ m/z=383.0 (M+H).
Ethyl 2-((5-bromo-2-methylphenyl)(isobutyl)amino)thiazole-4-carboxylate OR637-2 (method B, 60%) as a light yellow solid. Rf=0.34 (DCM); 1H NMR (400 MHz, CDCl3) δ 7.43 (dd, J=8.2, 2.0 Hz, 1H), 7.39 (d, J=2.0 Hz, 1H), 7.30 (s, 1H), 7.22 (d, J=8.2 Hz, 1H), 4.34 (q, J=7.1 Hz, 2H), 3.73 (brs, 2H), 2.17 (s, 3H), 2.05-1.92 (m, 1H), 1.36 (t, J=7.1 Hz, 3H), 0.98 (d, J=6.7 Hz, 6H); 13C NMR (100 MHz, CDCl3) δ 170.81, 161.85, 144.52, 143.86, 136.21, 133.84, 132.28, 131.96, 120.14, 117.00, 61.16, 59.87, 27.50, 20.51, 17.50, 14.46; LCMS C17H21BrN2O2S method (B) Rt=7.806 min, ESI+ m/z=397.0 (M+H).
Ethyl 2-((5-bromo-2-methylphenyl)(propyl)amino)oxazole-4-carboxylate OR0640-2. (method B, 60%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 7.67 (s, 1H), 7.38 (dd, J=8.2, 2.0 Hz, 1H), 7.33 (d, J=2.0 Hz, 1H), 7.16 (d, J=8.2 Hz, 1H), 4.35 (q, J=7.1 Hz, 2H), 3.81-3.73 (m, 2H), 2.12 (s, 3H), 1.66 (sext, J=7.4 Hz, 2H), 1.35 (t, J=7.1 Hz, 3H), 0.92 (t, J=7.4 Hz, 3H); LCMS C16H19BrN2O3 method (B) Rt=7.253 min, ESI+ m/z=367.0 (M+H).
Ethyl 2-((5-bromo-2-methylphenyl)amino)thiazole-4-carboxylate OR0614-2 (method A, quantitative) as a yellow solid, used in the next step without further purification. Rf=0.50 (PE-EtOAc, 70:30); 1H NMR (400 MHz, CDCl3) δ 7.66 (d, J=2.0 Hz, 1H), 7.49 (s, 1H), 7.23 (dd, J=8.1, 2.0 Hz, 1H), 7.11 (d, J=8.1 Hz, 1H), 4.29 (q, J=7.1 Hz, 2H), 2.23 (s, 3H), 1.33 (t, J=7.1 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 166.65, 161.45, 143.18, 139.75, 132.67, 129.64, 128.43, 124.36, 120.19, 117.02, 61.46, 17.59, 14.45; LCMS C13H13BrN2O2S method (B) Rt=6.664 min, ESI+ m/z=341.0 (M+H).
Ethyl 2-((5-bromo-2-methylphenyl)amino)oxazole-4-carboxylate OR0640-3 (method A, 21%) as a white solid. LCMS C13H13BrN2O3 method (B) Rt=6.396 min, ESI+ m/z=325.0 (M+H).
General Procedure for the Synthesis of 1-(5-halogeno-2-methylphenyl)thiourea Derivative. As previously described for synthesis of 1-(3-nitroaryl)thiourea.
1-(5-Iodo-2-methylphenyl)-1-propylthiourea OR0607-3 (98%) as a yellow solid, engaged in the next step without further purification. 1H NMR (400 MHz, CDCl3) δ 7.62 (dd, J=8.1, 1.7 Hz, 1H), 7.48 (d, J=1.8 Hz, 1H), 7.07 (d, J=8.1 Hz, 1H), 5.51 (brs, 2H), 4.38-0.30 (m, 1H), 3.64-3.56 (m, 1H), 2.18 (s, 3H), 1.80-1.55 (m, 2H), 0.90 (t, J=7.4 Hz, 3H); LCMS C11H15IN2S method (B) Rt=6.324 min, ESI+ m/z=335.0 (M+H).
1-(5-Bromo-2-methylphenyl)thiourea OR0614-3 (71%) as a yellow solid. Rf=0.18 (PE-EtOAc, 70:30); 1H NMR (400 MHz, MeOD) δ 7.42 (d, J=1.9 Hz, 1H), 7.38 (dd, J=8.2, 2.0 Hz, 1H), 7.21 (d, J=8.2 Hz, 1H), 2.23 (s, 3H); LCMS C8H9BrN2S method (B) Rt=4.773 min, ESI+ m/z=245.0 (M+H).
1-(5-Bromo-2-methylphenyl)urea OR0640-4
To a solution of 5-bromo-2-methylaniline (2.0 g, 10.75 mmol) in glacial acetic acid (22 mL) was added portionwise potassium cyanate (1.75 g, 21.5 mmol). The reaction mixture was stirred at r.t for 2 hours upon complete consumption of the starting material monitored by LCMS. The resulting suspension was diluted with water (200 mL) and the precipitate was collected by filtration affording 1-(5-bromo-2-methylphenyl)urea OR0640-4 (2.46 g, quantitative), as a beige solid, used in the next step without further purification. LCMS C8H9BrN2O method (B) Rt=4.833 min, ESI+ m/z=229.0 (M+H).
General Procedure for the Synthesis of N-((5-halogeno-2-methylphenyl)carbamothioyl)benzamide Derivative. As previously described for synthesis of N-([1,1′-biaryl]-3-ylcarbamothioyl)benzamide.
N-((5-Iodo-2-methylphenyl)(propyl)carbamothioyl)benzamide OR0607-4 (quantitative), as a yellow solid, used in the next step without further purification. LCMS C15H19IN2OS method (B) Rt=7.119 min, ESI− m/z=437.0 (M−H).
N-((5-Bromo-2-methylphenyl)carbamothioyl)benzamide OR0614-4 (quantitative) as a yellow solid, used in the next step without further purification. LCMS C15H13BrN2OS method (B) Rt=7.166 min, ESI+ m/z=349.0 (M+H).
5-Iodo-2-methyl-N-propylaniline OR0607-5
Using the same procedure as previously described for synthesis of [1,1′-biaryl]-3-N-alkylamine, (90%) as a light yellow solid, used in the next step without further purification. Rf=0.58 (EP-DCM, 70:30); 1H NMR (400 MHz, CDCl3) δ 6.95 (dd, J=7.7, 1.7 Hz, 1H), 6.89 (d, J=1.7 Hz, 1H), 6.75 (d, J=7.7 Hz, 1H), 3.51 (brs, 1H), 3.09 (t, J=7.3 Hz, 2H), 2.07 (s, 3H), 1.69 (sext, J=7.3 Hz, 2H), 1.03 (t, J=7.3 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 147.72, 131.56, 125.50, 121.30, 118.31, 92.36, 45.68, 22.70, 17.23, 11.77; LCMS C10H14IN method (B) Rt=7.538 min, ESI+ m/z=276.1 (M+H).
dCK protein contained three solvent exposed cysteines mutated to serines in order to generate better quality crystals (C9S, C45S, C59S) and a serine mutated to glutamic acid (S74E). The S74E mutation mimics the phosphorylated state of this serine, which favors the open conformation of the enzyme, making it competent for nucleoside binding to evaluate the binding affinity with compounds. BL21 pRIL Escherichia Coli cells were transformed with dCK-plasmid. Cells were grown in LB media containing 100 μg/ml ampicillin and 34 μg/ml chloramphenicol at 18° C. for 20 h after induction with 1 mM IPTG. Cells were re-suspended in 50 mM TRIS, pH 8, 500 mM NaCl, 30 mM imidazole and 10% glycerol buffer containing one EDTA free anti-protease tablet (Roche). After sonication and centrifugation (30,000×g), the supernatant was loaded on a 5 ml HisTrap FF column (GE healthcare) pre-equilibrated with resuspension buffer. Protein was then eluted by re-suspension buffer containing 500 mM imidazole. Protein eluted fractions were mixed and concentrated. Protein was further purified by size exclusion chromatography (superdex 200 16/600, GE healthcare) in 20 mM HEPES, pH 7.5, and 200 mM NaCl before concentration to 30 mg/ml and storage at −80° C.
Thermal Shift Assay (TSA) experiments were performed in triplicate in 384-well PCR white plates (Thermo Scientific, AB-3384) in the assay buffer containing 20 mM HEPES, pH 7.5 and 200 mM NaCl. Final concentrations were adjusted to 5 sM of protein, 50 sM of compound (final 2% DMSO) and dye diluted to 1:1000, as recommended by manufacturer (Protein Thermal Shift Dye, ThermoFisher Scientific). Controls with compounds alone or compounds with dye were performed to verify that compounds do not present autofluorescence. Plates were centrifuged at 1000 rpm for 2 min at 4° C. Thermal melting experiments were carried using a CFX-384 Connect RT-PCR (Bio-Rad). Plates were first equilibrated at 25° C.; then, the plates were heated from 25 to 95° C. by 0.5° C./min steps. Raw fluorescence data were treated, and the melting temperatures (Tm) were calculated using CFX Manager 3.1 software (Bio-Rad). ΔTm represents the difference in Tm between the protein in the presence of compound and the protein alone (both at 2% DMSO). Each experiment was performed at least twice (with technical triplicates) and data are presented as the mean f SD in Table 1.
The cellular assay was set up to assess the ability to inhibit dCK activity in a cellular model. The model was developed with a T-cell acute lymphoblastic leukemia cell line (CCRF-CEM), because of its high dependence on the salvage pathway, thus on dCK activity. Cellular assays were performed to assess the inhibition of cell proliferation under two conditions: i) in the presence of the molecule alone to determine the non-specific effect on cells, independent of dCK (non-specific toxicity) and ii) in the presence of an inhibitor of the De Novo pathway to determine the ability of molecules to inhibit dCK and decrease tumor proliferation (activity on dCK). The inhibitor of the De Novo pathway chosen for the experiments was thymidine (dT), a physiological inhibitor of ribonucleotide reductase which arrests cell proliferation in phase S. The dTTP produced via thymidine kinase from dT acts as a RNR inhibitor by an allosteric regulation of the R1 subunit (
CCFR-CEM cells were grown at 37° C. in RPMI 1640 with L-glutamine, supplemented with 100 units/ml penicillin, 100 mg/ml streptomycin, and 10% heat-inactivated foetal calf serum. The experiments were carried out on 96-well plates under the following conditions: 14,000 cells per well, RPMI 1640 culture medium supplemented with 10% heat-inactivated foetal calf serum, 200 sM of dT, 1 sM of dC and a range of dilutions of each molecule ranging from 40 μM to 10 μM, depending on the activity of the molecules (final 0.2% DMSO). Cells were incubated for 72 hours under these different conditions and finally, incubated with 10 μl/well of CellTiter-Blue reagent (Promega) for 4 hours at 37° C. The amount of reduced dye formed by metabolically active cells was quantified by its fluorescence measured at 560 nm (excitation) and 590 nm (emission) in a microplate reader (CLARIOstar, BMG Labtech). A blank well without cells was used as a background control. Each experiment was performed at least three times (with technical triplicates) and data are presented as the mean f SD.
All experiments were performed using standard methods in agreement with the French Guidelines for animal handling and approved by the local ethics committee APAFIS #6743. NOD-SCID-IL2R common-7-chain-knockout mice (NSG) were purchased from Charles River Laboratories and maintained under pathogen-free conditions on a 12-h light and 12-h dark cycle. Healthy 6 to 9 weeks-old male mice received 0.1×106 luciferase transduced-CCRF-CEM cells in the tail vein on Day 0 and were randomly assigned in four groups to receive intraperitoneal injections from Day 3 (Vehicle (n=8), dT (n=8), OR0642 (n=9) and dT+OR0642 (n=9)). Treatments were administered during 19 days at cycles of 5 days (BID) and 2 days (QD). Doses administered were: dT at 1.5 g/Kg, OR0642 at 40 mg/Kg and dT+OR0642 at 1.5 g/Kg and 40 mg/Kg, respectively. Vehicle was saline solution (0.9% NaCl) supplemented with 10% Kolliphor EL. OR0642 was administered in its hydrochloride salt form. Drug administration protocol is detailed in
Table 1 below shows results obtained with some compounds according to the invention for Affinity Assay (i.e., binding assay) via Thermal Shift Assay expressed in +ΔTm (°) and Cellular Assay (i.e., in vitro experiment on T-cell acute lymphoblastic leukemia cell line CCRF-CEM in presence of 200 μM dT and 1 μM dC) expressed in IC50 (nM).
The experimental evaluations by biochemical affinity assay and cellular assay obtained for 82 compounds are summarized in Table 1 and
Inventors developed a cellular assay to validate whether a combinatory inhibition of the De Novo and Salvage pathways by the combination of a dCK inhibitor with a Ribonucleotide Reductase (RNR) inhibitor could lead to the inhibition of cellular proliferation. Results on the human leukemia cell line CCRF-CEM (T-ALL) revealed that new compounds were able to inhibit cancer cells proliferation in combination with the physiologic RNR inhibitor dT in presence of dC. IC50 cell proliferation values were obtained between 37 μM and around 2 nM, more precisely between 37 μM and 1.6 nM (Table 1,
To investigate whether the efficacy observed in vitro could be correlated in vivo, compound OR0642 (hydrochlorhydre salt) was challenged in a systemic leukemia in vivo mice model. Inventors studied the in vivo efficacy and tolerability of co-targeting the alternative nucleotide biosynthetic pathways De Novo Pathway (DNP) and Salvage Pathway (SP) in a NOD-SCID-IL2Rγc null (NSG) mice model inoculated intravenously with luciferase expressing the human CCRF-CEM T-ALL cell line. The DNP was inhibited with dT, a physiologic inhibitor of RNR, and the SP with the newly developed dCK inhibitor OR0642. Treatment was initiated on day 3 after graft and was maintained during 19 days with cycles of 5 days (BID, twice/day) and 2 days (QD, once/day) (
| Number | Date | Country | Kind |
|---|---|---|---|
| 22305038.6 | Jan 2022 | EP | regional |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2023/050993 | 1/17/2023 | WO |
