EA040879B1 - Benzimidazole derivatives and their use as IDH1 inhibitors - Google Patents

Description

Cross-reference to related applications

This application takes precedence over CN application 201810672394.3 filed on June 26, 2018.

Field of invention

The present invention relates to a number of benzimidazole compounds and their use as mutant IDH1 (isocitrate dehydrogenase 1) inhibitors, in particular to a compound as represented by formula (I), a tautomer thereof, or a pharmaceutically acceptable salt thereof.

Prior Art

Isocitrate dehydrogenase is an important enzyme in the citric acid cycle and catalyzes the oxidative decarboxylation of isocitrate to 2-oxoglutarate (i.e. 2-a-ketoglutarate, α-KG). The protein encoded by the IDH1 gene is a NADP(+)-dependent isocitrate dehydrogenase containing the PTS-1 peroxysome targeting signal sequence present in the cytoplasm and peroxisomes. The presence of this enzyme in perokisosms indicates a role in the internal regeneration of NADPH.

A wild-type enzyme such as wild-type IDH catalyzes the oxidative decarboxylation of isocitrate with simultaneous reduction of NAD+(NADP+) to NADP(NADPH):

Isocitrate + NAD ⁺ (NADP ⁺ ) ^ α-KG + CO ₂ + NADP (NADPH) + H ⁺

Mutant protein IDH 1/2 (IDH 1/2 m) has been found in many different tumors, including glioma, acute myeloid leukemia (AML), chondrosarcoma, intrarenal cholangioma, melanoma, prostate cancer, angioimmunoblastic T-cell lymphoma. In gliomas, more than 70% of non-primary glioblastomas have IDH1 mutations, and 92.7% of tumors with mutant IDH1 have arginine substituted for histidine (i.e., IDH1 R132H) (Hartmann C, Acta Neuropathol. 2009 Oct; 1 18(4):469 -74).

The mutant IDH protein has a novel function, i.e. it catalyzes the reduction of α-KG to produce the carcinogenic metabolite 2-hydroxyglutarate (2-HG). The production of 2-HG is believed to contribute to the occurrence and development of cancer (Dang L, Nature, 2009 Dec 10;462(7274):739-44). Normal cells produce very low levels of 2-HG, while cells with IDH mutations are capable of producing high levels of 2-HG. High levels of 2-HG can also be found in tumors with IDH mutations.

Therefore, inhibition of mutant IDH and its novel activity is a potential way to treat cancer. Therefore, there is a need to develop IDH mutant inhibitors to inhibit 2-HG production.

Acta Neuropathol (2017, Vol(133), Issue 4, 629-644) discloses the specific structure of the compound BAY1436032.

OCF,

X™, 0

I || y—NH

4Λ _Ν

BAY1436032

Brief summary of the invention

The present invention provides a compound as represented in formula (I),

Where

R1 is selected from _{C1_6} alkyl, _{C3_6 cycloalkyl} and phenyl, with _{C1_6} alkyl, _{C3_6 cycloalkyl} and phenyl optionally substituted with 1, 2 or 3 R _a ;

each of R ₂ and R3 is independently selected from H, F, Cl, Br, I, OH, NH ₂ , CN, COOH, _{C1_6} alkyl and

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C1-6alkoxy, with C1-6alkyl and C1-6alkoxy optionally substituted with 1, 2 or 3 R _b ;

alternatively R1 and R2 are linked together to form a C _4-6 cycloalkenyl optionally substituted with 1 or 3 Rc;

L is -CH2CH2-;

n is selected from 1, 2 and 3; and each of Ra, R _b , and Rc is independently selected from F, Cl, Br, I, OH, NH2, CN, COOH, and Me;

or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

In some embodiments of the present invention, the aforementioned R1 is selected from C _1-3 alkyl, cyclopropanyl and phenyl, with C _1-3 alkyl, cyclopropanyl and phenyl optionally substituted with 1, 2 or 3 Ra, and the remaining variables are as defined in the present description of the invention.

In some embodiments of the present invention, the aforementioned R1 is selected from

CH3, CH ₂ F, CHF ₂ , CF3, CH ₂ CH3, CH ₂ CF ₃ , CH ₂ CH 2 _CH3 , C(CH3)3, V, CH(CH3) ₂ and

and the remaining variables are as defined in the present description of the invention.

In some embodiments of the present invention, each of the above R2 and _R3 is independently selected from H, F, Cl, Br, I, OH, NH2, CN, COOH, C _1-3 alkyl and C _1-3 alkoxy, wherein C _1-3 alkyl and C1 ₃ alkoxy are optionally substituted with 1, 2 or 3 R _b , and the remaining variables are as defined herein.

In some embodiments of the present invention, each of the above R2 and R3 is independently selected from H, F, Cl, Br, I, OH, NH2, CN, COOH, CH ₃ and CH3O, with CH ₃ and CH3O optionally substituted with 1, 2 or 3 R _b , and the remaining variables are as defined in the present description of the invention.

In some embodiments of the present invention, each of the above R2 and _R3 is independently selected from H, F, Cl, Br, I, OH, NH2, CN, COOH, _CH3 , CH2F, CHF2, _CF3, and _OCH3 , and the remaining variables are , as defined in the present description of the invention.

In some embodiments of the present invention, the aforementioned L is CH2CH2- and the remaining variables are as defined in the present description of the invention.

In some embodiments of the present invention, the above structural

p * R3 T/G ^2 selected from ^{and z}

’ and the rest of the variables are as defined by unity in the present unit specification.

In some embodiments of the present invention, the above structural

selected from the rest of the variables as defined in the present description of the invention.

A

other variables are as defined in the present description of the invention.

In some embodiments of the present invention, the above structural unit

selected from

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and the remaining variables are as defined in the present description of the invention.

There are also some embodiments of the present invention obtained with any combination of the above variables.

In some embodiments of the present invention, the aforementioned compound, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from

Where

E is selected from -CH ₂ -, -CH2CH2-, -C(CH ₃ ) ₂ - and -C(CH ₃ ) ₂ CH ₂ -;

L is -CH2CH2-; And

R1, R2 and R3 are as defined in the present specification.

In some embodiments of the present invention, the aforementioned compound, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from

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Where

E, L, R1, R2 and R3 are as defined in the present specification.

The present invention also provides a compound as represented in the following formula, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, which is selected from

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In some embodiments of the present invention, the aforementioned compound is selected from

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The present invention also provides the use of the aforementioned compound, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a disease associated with IDH1 (isocitrate dehydrogenase 1).

Definition and description of the invention

Unless otherwise noted, the following terms and phrases used in this document have the following meanings. A specific term or a specific phrase for which no specific definition has been given should not be regarded as indefinite (indefinite) or obscure (obscure), but should be understood in its usual meaning. When a trade name appears in this specification, it means that it refers to the respective product or its active ingredient.

The term "pharmaceutically acceptable" as used herein refers to compounds, substances, compositions and/or dosage forms which, within the reasonable medical judgment, are suitable for contact with human and animal tissues without undue toxicity, irritation, allergic reactions, or other problems or complications, and which commensurate with a reasonable benefit/risk ratio.

The term pharmaceutically acceptable salt refers to a salt of a compound of the present invention that is derived from a compound with the specific substituents disclosed in the present invention and a relatively non-toxic acid or base. When a compound of the present invention contains a relatively acidic functional group, then a base addition salt can be prepared by contacting the neutral form of the compound with a specific

- 7 040879 an amount of base in pure solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, organic ammonium or magnesium salts or the like. When a compound of the present invention contains a relatively basic functional group, then an acid addition salt can be prepared by contacting the neutral form of the compound with a sufficient amount of acid in pure solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid addition salts including, for example, hydrochloride, hydrobromide, nitrate, carbonate, bicarbonate, phosphate, monohydrogen phosphate, dihydrogen phosphate, sulfate, hydrogen sulfate, hydroiodide, phosphite, etc., as well as organic acid addition salts. acids, where the organic acid includes, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanedic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid acid, citric acid, tartaric acid and methanesulfonic acid, etc.; and also includes salts of amino acids (such as arginine, etc.), and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functionalities and can therefore be converted to either a base addition salt or an acid addition salt.

The pharmaceutically acceptable salt of the present invention can be synthesized from the parent compound containing an acidic or basic functional group, by conventional chemical methods. Typically, such salts are prepared by reacting these compounds in free acid or free base form with stoichiometric amounts of suitable bases or acids in water or in organic solvents, or a mixture of both.

The compounds of the present invention may exist in particular geometric or isomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)isomers and their racemic mixture; and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present on a substituent such as alkyl. All of these isomers and mixtures thereof are within the scope of the present invention.

Unless otherwise noted, the term enantiomer or optical isomer refers to isomers that are mirror images of each other.

Unless otherwise indicated, the term cis-trans isomer or geometric isomer is due to the inability to rotate freely due to the double bonds or single bonds of the carbon atoms forming the ring.

Unless otherwise indicated, the term diastereomer refers to stereoisomers that have two or more chiral centers and whose molecules are not mirror images of each other.

Unless otherwise indicated, (D) or (+) means right hand rotation, (L) or (-) means left hand rotation, and (DL) or (±) means racemic.

Unless otherwise indicated, wedge-shaped solid line ) and wedge-shaped dotted line (straight dashed line) are used to indicate relative stereocenter configuration, are used to indicate absolute stereocenter configuration, straight solid line ν) and straight dashed line (') are used to indicate relative stereocenter configuration, a wavy line ( ) is used to indicate a wedge-shaped solid line (*^ ) or a wedge-shaped dotted line ('' ), a wavy line ) is used to indicate a straight solid line (^ ) and a straight dashed line )'

Unless otherwise noted, for a compound in which there is a double bond, such as a carbon-carbon double bond, a carbon-nitrogen double bond, and a nitrogen-nitrogen double bond, and each atom on the double bond is bonded to two different substituents (in a double bond containing an atom nitrogen, a single pair of electrons on the nitrogen atom is considered a substituent associated with it); if an atom on a double bond in a compound is bonded to a compound substituent with a wavy line L, this means the (Z) isomer, (E) isomer, or a mixture of two isomers of the compound. For example, the following formula (A) means that the compound exists as a single isomer of formula (A-1) or formula (A-2), or as a mixture of two isomers of formula (A-1) and formula (A-2). The following formula (B) means that the compound exists as a single isomer of formula (B-1) or formula (B-2), or as a mixture of two isomers of formula (B-1) and formula (B-2). The following formula (C) means that the compound exists as a single isomer of formula (C-1) or formula (C-2), or as a mixture of two isomers of formula (C-1) and formula (C-2).

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The compounds of the present invention may exist in specific forms. Unless otherwise indicated, the term tautomer or tautomeric form means that at room temperature, isomers with different functional groups are in dynamic equilibrium and can quickly turn into each other. If tautomers are possible (such as in solution), a chemical equilibrium of the tautomers can be achieved. For example, a proton tautomer, also referred to as a prototropic tautomer, includes interconversion via proton migration such as keto-enol isomerization and imine-enamine isomerization. The valence tautomer involves some rearrangement of bond electrons for interconversion. A specific example of keto-enol tautomerism is the tautomerism between the two tautomers pentan-2,4-dione and 4-hydroxypent-3-en-2-one.

Unless otherwise indicated, the terms enriched in one isomer, enriched in isomer, enriched in one enantiomer, or enantiomeric enrichment means that the content of one isomer or enantiomer is less than 100%, and the content of an isomer or enantiomer is 60% or more, or 70% or more, or 80% or more, or 90% or more, or 95% or more, or 96% or more, or 97% or more, or 98% or more, or 99% or more, or 99.5% or more, or 99.6% or more, or 99.7% or more, or 99.8% or more, or 99.9% or more.

Unless otherwise indicated, the term isomeric excess or enantiomeric excess refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, then the isomeric or enantiomeric excess (ee value) is 80%.

Optically active (R)- and (S)-isomers and D and L isomers can be obtained by chiral synthesis or using chiral reagents or other standard methods. To obtain the enantiomer of the compound of the present invention, it can be obtained by asymmetric synthesis or by derivatization with chiral auxiliary agents, in which the resulting mixture of diastereomers is separated and the auxiliary group is cleaved off to obtain the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), it can form diastereomeric salts with suitable optically active acids or bases, and the diastereomers can then be separated by standard methods known in the art. to isolate pure enantiomers. In addition, the separation of enantiomers and diastereomers is usually carried out by chromatography, which uses a chiral stationary phase, and possibly in combination with chemical derivatization (for example, carbamate is formed from an amine). The compounds of the present invention may contain unnatural ratios of atomic isotopes on one or more of the atoms that make up the compound. For example, compounds can be labeled with radioisotopes such as tritium ( ³ H), iodine-125 ( ¹²⁵ I) or C-14 ( ¹⁴ C). As another example, deuterated drugs can be formed by replacing hydrogen with deuterium. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared to non-deuterated drugs, deuterated drugs have reduced toxic side effects and improved drug stability, improved potency, longer biological half-life, and other advantages. All changes in the isotopic composition of the compounds of the present invention, whether radioactive or not, are within the scope of the present invention.

Possible or possible means that the event or condition described below may not necessarily occur, and the description covers both the situation where the event or condition occurs and the situation where the event or condition does not occur.

The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by substituents, including deuterium and hydrogen variants, as long as the valency of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (ie =O), then it means that two hydrogen atoms have been substituted. Oxygen substitution does not occur on aromatic groups. The term possibly substituted means that

- 9 040879 the group may be substituted or unsubstituted. Unless otherwise indicated, the type and number of substituents may be arbitrary, as long as they can be chemically implemented.

When any variable (such as R) occurs more than once in the composition or structure of a compound, then its definition in each case is independent. Thus, for example, if a group is substituted with 0-2 R, then that group may possibly be substituted with up to two R's, and in each case R is independently selected. In addition, combinations of substituents and/or variants thereof are only permitted as long as such combinations result in stable compounds.

When the number of linker groups is 0, for example -(CRR) ₀ -, then it means that the linker group is a single bond.

When one of the variables is selected from a single link, then it means that the two groups are directly related. For example, when the L in A-L-Z is a single bond, then this means that the structure is actually A-Z.

When a substitute is not used (is vacant), then this means that this substitute is not available. For example, when X in A-X is not used, then this means that the structure is really A. When it is not specifically indicated through which atom a listed substituent is bonded to a substituted group, then such substituents may be bonded at any atom. For example, a pyridyl group as a substituent may be linked to a substituted group through any carbon atom on the pyridine ring. When the direction of linkage of the listed linker groups is not specifically indicated, the direction of linkage is arbitrary.

For example, if the linker group L in represents -M-W-, then -M-W- can link ring A and ring B in the direction in reading order from left to right to form

and can also link ring A and ring B in the opposite direction in

reading order from right to left with education

Combinations of linker groups, substituents, and/or variants thereof are permitted as long as such combinations result in stable compounds.

Unless otherwise indicated, the term C _1-6 alkyl is used to indicate a linear or branched saturated hydrocarbon group of 1-6 carbon atoms. The C _1-6 alkyl group includes C _1-5 , C _1-4 , C _1-3 , C _1-2 , C _2-6 , C ₂ -4 , C ₆ and C 5 alkyl groups, etc. and may be monovalent (such as methyl), divalent (such as methylene), or multivalent (such as methine). Examples of Cp ₆ alkyl groups include, without limitation, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, sec-butyl, and tert-butyl ), pentyl (including n pentyl, isopentyl and neopentyl), hexyl, etc.

Unless otherwise indicated, the term C _1-3 alkyl applies to a linear or branched saturated hydrocarbon group of 1-3 carbon atoms. The C _1-3 alkyl group includes C _1-2 and C _2-3 alkyl groups, and so on. and may be monovalent (such as methyl), divalent (such as methylene), or multivalent (such as methine). Examples of C _1-3 alkyl groups include, without limitation, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

Unless otherwise indicated, the term C _1-6 alkoxy refers to an alkyl group containing 1-6 carbon atoms, which is attached to the rest of the molecule through an oxygen atom. The C _1-6 alkoxy group includes C _1-4 , C _1-3 , C _1-2 , C _2-6 , C _2-4 , C ₆ , C ₄ , C ₄ , C ₃ alkoxy, etc. Examples of C _1-6 alkoxy include, without limitation, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, sec-butoxy and tert-butoxy), pentoxy (including n-pentoxy, isopentoxy and neopentoxy), hexoxy and the like.

Unless otherwise indicated, the term C _1-3 alkoxy refers to an alkyl group containing 1-3 carbon atoms that are attached to the rest of the molecule through an oxygen atom. The C _1-3 alkoxy group includes C _1-2 , C _2-3 , C ₃ and C ₂ alkoxy groups, etc. Examples of C _1-3 alkoxy groups include, without limitation, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

Unless otherwise indicated, C _3-6 cycloalkyl means a saturated cyclic hydrocarbon group of 3-6 carbon atoms which is a monocyclic and bicyclic ring system. C _3-6 cycloalkyl includes C _3-5 , C _4-5 and C _5-6 cycloalkyl, etc. and may be monovalent, divalent or multivalent. Examples of C _3-6 cycloalkyl groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

Unless otherwise indicated, C _4-6 cycloalkenyl means a partially unsaturated cyclic hydrocarbon group consisting of 4-6 carbon atoms, containing at least one carbon-carbon double bond, and which is a monocyclic and bicyclic system,

- 10 040879 where the bicyclic system includes a spiro ring, a fused ring and a bridged ring, in which each ring is non-aromatic. C _4-6 cycloalkenyl group includes C _4-5 or

C _5-6 cycloalkenyl group, etc. and may be monovalent, divalent or multivalent.

Examples of C _4-6 cycloalkenyl include, without limitation, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, and the like.

Unless otherwise indicated, the terms C _6-12 aromatic ring and C _6-12 aryl may be used interchangeably herein. The term C _6-12 aromatic ring or C _6-12 aryl means a cyclic hydrocarbon group of 6-12 carbon atoms with a conjugated π electron system, which may be a single ring, a fused bicyclic ring or a fused tricyclic ring system, where each ring is aromatic. It can be monovalent, divalent or multivalent. C _6-12 aryl groups include C _6-10 , C _6-9 , C _6-8 , C ₁₂ , Xu and C ₆ aryl groups. Examples C ₆ . _{The 12} aryl groups include, without limitation, phenyl, naphthyl (including 1-naphthyl, 2-naphthyl, etc.).

Unless otherwise indicated, C _n -n ₊ m or C _n - _C n ₊ m includes any particular case from n to n+m carbon atoms; for example, C _1-12 covers C1, C2, C ₃ , C ₄ , C5, C ₆ , C ₇ , C ₈ , C ₉ , C ₁₀ , C11 and C ₁₂ , including any range from n to n+m ; for example, C _1-12 covers C _1-3 , C _1-6 , C _1-9 , C _3-6 , C _3-9 , C _3-12 , C _6-9 , C _6-12 and C _{9- 12} etc. Similarly, from n-membered to n+m-membered means that the number of atoms in the ring is from n to n+m; for example, 3-12 membered ring includes 3-membered ring, 4-membered ring, 5-membered ring, 6-membered ring, 7-membered ring, 8-membered ring, 9-membered ring, 10-membered ring, 11-membered ring ring and 12-membered ring, including any range from n to n+m; for example, a 3-12 membered ring includes a 3-6 membered ring, a 3-9 membered ring, a 5-6 membered ring, a 5-7 membered ring, a 6-7 membered ring, a 6-8 membered ring and a 6 to 10 membered ring, and the like.

The term leaving group refers to a functional group or atom that can be replaced by another functional group or atom in a substitution reaction (eg, an affine substitution reaction). For example, representative leaving groups include triflat; chlorine, fluorine, bromine, iodine; a sulfonate such as mesylate, tosylate, p-bromobenzenesulfonate, p-toluenesulfonate, etc.; acyloxy such as acetoxy, trifluoroacetoxy, etc.

The term protecting group includes, without limitation, an amino protecting group, a hydroxy protecting group, or a thiol protecting group. The term amino protecting group refers to a protecting group suitable for preventing side reactions at the nitrogen position of the amino group. Representative amino protecting groups include, without limitation: formyl; acyl such as alkanoyl (such as acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl such as t-butoxycarbonyl (Boc); arylmethyloxycarbonyl such as benzyloxycarbonyl (Cbz) and 9fluorenylmethyloxycarbonyl (Fmoc); arylmethyl such as benzyl (Bn), trityl (Tr), 1,1-di-(4'methoxyphenyl)methyl; silyl such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc. The term hydroxyl protecting group refers to a protecting group suitable for preventing side reactions of the hydroxyl group. Representative hydroxyl protecting groups include, without limitation: alkyl groups such as methyl, ethyl, and tert-butyl; acyl groups such as alkanoyl (such as acetyl); arylmethyl such as benzyl (Bn), p-methyloxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc.

The compounds of the present invention may be prepared by various synthetic routes well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by their combinations with other chemical synthesis methods, and their equivalents or alternatives known to those skilled in the art. Preferred embodiments include, without limitation, the examples given in the present description of the invention.

The solvent used in the present invention is commercially available. In the present description of the invention, the following abbreviations are used: aq. means water; HATU is O-(7azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; EDC means N-(3dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride; m-CPBA means 3-chloroperoxybenzoic acid; equiv. means equivalent; CDI means carbonyldiimidazole; DCM means dichloromethane; PE means petroleum ether; DIAD means diisopropyl azodicarboxylate; DMF means N,Ndimethylformamide; DMSO means dimethyl sulfoxide; EtOAc means ethyl acetate; EtOH means ethanol; MeOH means methanol; CBz means benzyloxycarbonyl, which is the protecting group for the amino group; BOC means tert-butoxycarbonyl, which is an amino protecting group; HOAc means acetic acid; NaCNBH ₃ is sodium cyanoborohydride; k.t. means room temperature; O/N means during the night; THF means tetrahydrofuran; Boc ₂ O is di-tert-butyl dicarbonate; TFA means trifluoroacetic acid; DIPEA means diisopropylethylamine; SOC12 means thionyl chloride; CS2 means carbon disulfide; TsOH means p-toluenesulfonic acid; NFSI means N-fluoro-N-(benzenesulfonyl)benzenesulfonamide; NCS means 1-chloropyrrolidine-2,5-dione; p-Bu ₄ NF means tetrabutylammonium fluoride; iPrOH means 2 propanol; Tm means melting point; EDCl is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.

Compounds are named manually or using ChemDraw® software, and commercially available compounds are listed under the supplier's catalog names.

EFFECT: at the enzymatic level, the compound of the present invention has a good inhibitory effect on the mutant IDH1R132H and IDH1R132C, and at the same time does not have an inhibitory effect on the wild-type IDH protein; at the cellular level, the compound of the present invention has a good 2-HG inhibitory effect on U87MG glioma cells with an IDH1R132H mutation; the compound of the present invention has a significant inhibitory effect and good selectivity for mutant IDH1, and at the same time has a better distribution coefficient in brain tumors and pericerebral tissues, which can reduce potential side effects on normal brain tissues.

Detailed description of the invention

The following examples are intended to describe the present invention in detail, but are not intended to limit the present invention in any way. The present invention is described in detail here, and its specific embodiments are also disclosed. For a person skilled in the art it will be obvious that various changes and improvements can be made in relation to specific embodiments of the present invention without deviating from the intent and scope of the present invention.

Method A.

where the groups are as defined in the compound of formula (I) of the present invention. Example 1

Example 1A.

Potassium carbonate (2.46 g, 17.76 mmol) was added to a solution of methyl-(E)-3-(4-fluoro-3-nitrophenyl)acrylate (2 g, 8.88 mmol) and (R)-1- phenylethylamine (1.18 g, 9.77 mmol, 1.26 ml) in tetrahydrofuran (20 ml), and the mixed solution was stirred at 50°C for 16 hours. The reaction solution was diluted with 50 ml of water and then extracted with ethyl acetate (30 ml x ). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. At 15°C, the crude product was stirred in petroleum ether:ethyl acetate=10:1 (11 ml) for 30 min, filtered, and the solid was concentrated under reduced pressure to obtain the compound of example 1A. Ή NMR (400 MHz, CDCl ₃ ) δ=8.65 (d, J=5.4 Hz, 1H), 8.35 (d, J=2.1 Hz, 1H), 7.60-7.44 (m, 2H), 7.42- 7.29 (m, 5H), 6.70 (d, J=8.9Hz, 1H), 6.27 (d, J=15.9Hz, 1H), 4.76 (q, J=6.5Hz, 1H), 3.80 (s, 3H), 1.70 (d, J=6.7 Hz, 3H).

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Example 1B.

Under the protection of nitrogen gas, nickel chloride (1.99 g, 15.32 mmol) was added to a solution of the compound of example 1A (1 g, 3.06 mmol) in methanol (20 ml), after cooling to 0°C, sodium borohydride ( 695.57 mg, 18.39 mmol) and the mixed solution was stirred at 15° C. for 1 hour. The reaction mixture was quenched by adding 100 ml of saturated ammonium chloride solution and stirred for 15 minutes. The reaction solution was extracted with ethyl acetate (80 mlx3) and the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give Example 1B, which was directly used in the next step without purification.

Example 1C.

4-(Trifluoromethoxy)phenylisothiocyanate (888.84 mg, 4.06 mmol) was added to a solution of the compound of Example 1B (1.1 g, 3.69 mmol) in tetrahydrofuran (20 ml), stirred at 40° C. for half an hour, then EDC-HCl (1.41 g, 7.37 mmol) was added and stirred at 70°C for 16 hours. The reaction solution was diluted with 30 ml of water and then extracted with ethyl acetate (30 mlx3). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product which was purified by column chromatography to give Example 1C. 1Н NMR (400 MHz, CDCl ₃ ) δ=7.43-7.29 (m, 6H), 7.21-7.17 (m, 2H), 7.09-6.98 (m, 3H), 6.91 (br d, J=7.9 Hz, 1H), 5.80 (br s, 1H), 5.67 (q, J=7.1 Hz, 1H), 3.70-3.48 (m, 3H), 2.98 (t, J=7.9 Hz, 2H), 2.61 ( t, J=7.9 Hz, 2H), 1.82(d, J=7.1 Hz, 3H).

Example 1

Lithium hydroxide (334.35 mg, 13.96 mmol) was added to a solution of the compound of example 1C (1.35 g, 2.79 mmol) in tetrahydrofuran (10 ml) and water (10 ml), stirred at 15°C for 16 h, the reaction solution was brought to pH 6 with 1M hydrochloric acid. The reaction solution was extracted with ethyl acetate (30 ml x 3) and the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product. The crude product was diluted with 5 ml of ethyl acetate and then 20 ml of petroleum ether was added to precipitate a white solid. The solid was filtered off, concentrated under reduced pressure and then diluted with 10 ml of methyl tert-butyl ether, stirred for ten minutes, filtered and the solid was concentrated under reduced pressure to give the compound of example 1. 1H NMR (400 MHz, methanol^) δ=7.57 (br d, J=8.9 Hz, 2H), 7.43-7.21 (m, 8H), 6.90-6.63 (m, 2H), 5.97 (br d, J=6.8 Hz, 1H), 3.02-2.83 (m, 2H), 2.59 (br t, J=7.0 Hz, 2H), 2.03-1.92 (m, 3H)

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Example 2

Example 2A.

Potassium carbonate (2.46 g, 17.76 mmol) was added to a solution of methyl-(E)-3-(4-fluoro-3-nitrophenyl)acrylate (2 g, 8.88 mmol) and (S)-1-phenylethylamine (1.08 g, 8.88 mmol) in tetrahydrofuran (20 ml), the mixed solution was stirred at 50° C. for 16 hours. The reaction solution was diluted with 50 ml of water and then extracted with ethyl acetate (30 ml×3). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. At 15°C, the crude product was stirred in petroleum ether:ethyl acetate=10:1 (11 ml) for 30 min, filtered, and the solid was concentrated under reduced pressure to obtain the compound of example 2A. 1H NMR (400 MHz, CDCl ₃ ) δ=8.65 (br d, J=5.5 Hz, 1H), 8.35 (d, J=2.1 Hz, 1H), 7.56 (d, J=15.9 Hz , 1H), 7.47 (dd, J=2.0, 8.9 Hz, 1H), 7.41-7.29 (m, 5H), 6.70 (d, J=9.0 Hz, 1H), 6.27 (d, J=15 .9 Hz, 1H), 4.76 (q, J=6.5 Hz, 1H), 3.80 (s, 3H), 1.70 (d, J=6.7 Hz, 3H).

Example 2B.

Under the protection of nitrogen gas, nickel chloride (1.99 g, 15.32 mmol) was added to a solution of the compound of example 2A (1 g, 3.06 mmol) in methanol (20 ml), after cooling to 0°C, sodium borohydride ( 695.57 mg, 18.39 mmol), the mixed solution was stirred at 15° C. for 1 hour. The reaction mixture was quenched by adding 100 ml of saturated ammonium chloride solution and stirred for 15 minutes. The reaction solution was extracted with ethyl acetate (80 mlx3), the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give Example 2B, which was directly used in the next step without purification.

Example 2C.

4-(Trifluoromethoxy)phenylisothiocyanate (767.64 mg, 3.50 mmol) was added to a solution of the compound of Example 2B (950.00 mg, 3.18 mmol) in tetrahydrofuran (2 ml), stirred at 40° C. for half an hour, then EDCl (1.22 g, 6.37 mmol) was added and stirred at 70°C for 16 hours. The reaction solution was diluted with 30 ml of water and then extracted with ethyl acetate (30 mlx3). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product which was purified by column chromatography to give Example 2C. 1Н NMR (400 MHz, CDCl ₃ ) δ=7.43-7.27 (m, 6H), 7.23-7.17 (m, 2H), 7.07-6.97 (m, 3H), 6.95-6.86 (m, 1H), 5.82 (br s, 1H), 5.67 (q, J=7.0 Hz, 1H), 3.61 (s, 3H), 2.97 (t, J=7.9 Hz, 2H), 2.61 (t, J=7.9 Hz , 2H), 1.82 (d, J=7.1 Hz, 3H).

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Example 2

Lithium hydroxide (329.42 mg, 13.75 mmol) was added to a mixed solution of the compound of Example 2C (1.33 g, 2.75 mmol) in tetrahydrofuran (10 ml) and water (10 ml), stirred at 15°C in over 16 h, the reaction solution was brought to pH 6 with 1M hydrochloric acid. The reaction solution was extracted with ethyl acetate (30 mlx3), the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was diluted with 5 ml of ethyl acetate and then 20 ml of petroleum ether was added to precipitate a white solid. The solid was filtered off, concentrated under reduced pressure, then diluted with 10 ml methyl t-butyl ether, stirred for ten minutes, filtered, and the solid was concentrated under reduced pressure to give the compound of example 2. 1H NMR (400 MHz, methanol^) δ=7.56 (br d, J=7.2 Hz, 2H), 7.42-7.22 (m, 8H), 6.866.71 (m, 2H), 6.03-5.88 (m, 1H), 2.98-2.87 (m, 2H), 2.63-2.53 (m, 2H), 2.05-1.92 (m, 3H).

Example 3

OCF ₃

A O

Example 3A.

^4O a

LI

NH

AA

Potassium carbonate (982.04 mg, 7.11 mmol) and (1R)-1-(4-chlorophenyl)ethylamine (552.91 mg, 3.55 mmol) were added to a solution of (E)-3-(4 -fluoro-3-nitro-phenyl)prop-2-enoic acid (800 mg, 3.55 mmol) in tetrahydrofuran (15 ml) and this mixture was stirred at 45° C. for 20 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with 100 ml water and extracted with 300 ml (100 mlx3) ethyl acetate. The combined organic layer was washed with 200 ml (100 ml×2) saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography to give the compound of Example 3A. 1H NMR (400 MHz, CDCl ₃ ) δ=8.58 (br d, J=5.4 Hz, 1H), 8.34 (d, J=2.1 Hz, 1H), 7.55 (d, J=16.0 Hz , 1H), 7.47 (dd, J=2.1, 9.0 Hz, 1H), 7.367.32 (m, 2H), 7.29-7.27 (m, 2H), 6.62 (d, J=9.0 Hz, 1H ), 6.27 (d, J=16.0Hz, 1H), 4.71 (quin, J=6.4Hz, 1H), 3.79 (s, 3H), 1.66 (d, J=6.7Hz, 3H) .

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Example 3B.

To a solution of the compound of example 3A (1.01 g, 2.80 mmol) in methanol (10 ml) and tetrahydrofuran (6 ml) was added nickel chloride (3.33 g, 14.00 mmol) and then sodium borohydride solution was added dropwise (1.38 g, 36.39 mmol) in dimethylformamide (3 ml), making sure that the internal temperature did not exceed 10°C, then the reaction mixture was stirred at 10°C for 1 hour. Water (80 ml) and ethyl acetate (30 ml) was added to the mixture and filtered through a celite filter. The filter cake was washed with ethyl acetate (40 mlx3), the filtrate was extracted with 150 ml of ethyl acetate. The combined organic phase was washed with saturated brine (100 ml x 2), dried over anhydrous sodium sulfate and concentrated in vacuo to give Example 3B, which was used directly in the next step without purification.

Example 3C.

1-Isothiocyanato-4-(trifluoromethoxy)benzene (515.06 mg, 2.35 mmol, 381.52 µl) was added to a solution of Example 3B (711 mg, 2.14 mmol) in tetrahydrofuran (8 ml) and this the mixture was stirred at 45° C. for 1 h, then EDCl (409.53 mg, 2.14 mmol) was added and the mixture was stirred at 70° C. for 15 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with 50 ml water and extracted with 100 ml (50 ml x 2) ethyl acetate. The combined organic layer was washed with 100 ml (50 mlx2) saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0 to 10:1) to give Example 3C, which was directly used in the next step without purification.

Example 3

Lithium hydroxide (288.84 mg, 6.88 mmol) was added to a solution of the compound of Example 3C (713 mg, 1.38 mmol) in tetrahydrofuran (4 ml) and water (2 ml), this mixture was stirred at 20°C for 16 h. The mixture was acidified to pH=6 with dilute hydrochloric acid (1N) and extracted with 200 ml (100 mlx2) of ethyl acetate. The combined organic layer was washed with 200 ml (100 mlx2) saturated aqueous brine, dried using anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by high performance liquid chromatography (TFA conditions) to give Example 3D. 1Н NMR (400 MHz, DMSO-d ₆ ) δ=7.78 (br s, 2H), 7.47 (s, 6H), 7.26 (s, 1H), 6.98-6.76 (m, 2H), 6.14-6.03 (m, 1H), 3.42 (br s, 2H), 2.85 (br t, J=7.3 Hz, 2H), 1.96 (br d, J=6.7 Hz, 3H).

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Example 4

Example 4A.

Potassium carbonate (982.04 mg, 7.11 mmol) and (1R)-1-(4-fluorophenyl)ethylamine (494.45 mg, 3.55 mmol) were added to a solution of (E)-3-(4 -fluoro-3-nitro-phenyl)prop-2-enoic acid (800 mg, 3.55 mmol) in tetrahydrofuran (15 ml) and this mixture was stirred at 45° C. for 20 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with 100 ml water and extracted with 300 ml (100 mlx3) ethyl acetate. The combined organic layer was washed with 200 ml (100 mlx2) of brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0 to 5:1) to give the compound of Example 4A. 1H NMR (400 MHz, CDCl ,) δ=8.59 (br d, J=5.1 Hz, 1H), 8.34 (d, J=2.2 Hz, 1H), 7.55 (d, J=15.9 Hz, 1H), 7.48 (dd, J=2.1, 8.9 Hz, 1H), 7.34-7.28 (m, 2H), 7.09-7.01 (m, 2H), 6.65 (d, J=8.9 Hz, 1H), 6.27 (d, J=16.0Hz, 1H), 4.73 (quin, J=6.5Hz, 1H), 3.79 (s, 3H), 1.66 (d, J=6.7Hz, 3H ).

Example 4B.

To a solution of the compound of Example 4A (1.01 g, 2.93 mmol) in methanol (6 ml) and tetrahydrofuran (6 ml) was added nickel chloride (3.49 g, 14.67 mmol) and then sodium borohydride solution was added dropwise (1.4 g, 38.1 mmol) in dimethylformamide (3 ml), making sure that the internal temperature does not exceed 10°C, and then the reaction mixture was stirred at 10°C for 1 . The mixture was poured into water (80 ml) and ethyl acetate (30 ml) and filtered through a celite filter. The filter cake was washed with ethyl acetate (40 mlx3), the mixture was extracted with 150 ml of ethyl acetate. The combined organic phase was washed with saturated brine (100 ml x 2), dried over anhydrous sodium sulfate and concentrated in vacuo to give Example 4B, which was used directly in the next step without purification.

Example 4C.

1-Isothiocyanoyl-4-(trifluoromethoxy)benzene (313.22 mg, 1.43 mmol) was added to a solution of Example 4B (411 mg, 1.30 mmol) in tetrahydrofuran (6 mL) and this mixture stirred at 45° C. for 1 h, then EDCl (249.04 mg, 1.30 mmol) was added and the mixture was stirred at 70° C. for 15 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with 50 ml water and extracted with 100 ml (50 ml x 2) ethyl acetate. The combined organic layer was washed with 100 ml (50 ml x 2) of an aqueous solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography to obtain the compound of Example 4C, which was directly used in the next step without purification.

Example 4

Lithium hydroxide (139.33 mg, 3.32 mmol) was added to a solution of the compound of Example 4C (333 mg, 664.05 µmol) in tetrahydrofuran (4 ml) and water (2 ml) and the mixture was stirred at 20°C in for 16 hours. The mixture was acidified with dilute hydrochloric acid (1N, 6 ml) to pH=6 and extracted with 200 ml (100 mlx2) of ethyl acetate. The combined organic layer was washed with 200 ml (100 ml x 2) of brine and dried using anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by high performance liquid chromatography (TFA conditions) to give the compound of example 4. 1H NMR (400 MHz, DMSO-d ₆ ) δ=7.79 (br d, J=8.4 Hz, 2H), 7.49 (br s, 4H) , 7.29-7.21 (m, 3H), 6.95-6.78 (m, 2H), 6.08 (br d, J=7.0 Hz, 1H), 3.44-3.39 (m, 2H), 2.84 (br t, J= 7.4 Hz, 2H), 1.96 (br d, J=6.7 Hz, 3H).

Example 5

Example 5A.

Potassium carbonate (920.67 mg, 6.66 mmol) and (1R)-1-phenylprop-1-amine (330.25 mg, 2.44 mmol, 350.58 µl) were added to the methyl-(E)- 3-(4-fluoro-3-nitro-phenyl)acrylate (500 mg, 2.22 mmol) in tetrahydrofuran (10 ml) and this mixture was stirred at 50° C. for 16 h. The reaction mixture was diluted with water (20 ml ) and then extracted with ethyl acetate (10 mlx3). The combined organic layer was washed with brine (30 ml), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give Example 5A, which was used directly in the next step without purification. 1H NMR (400 MHz, CDCl ₃ ) δ=8.67 (br d, J=5.8 Hz, 1H), 8.24 (d, J=2.1 Hz, 1H), 7.46 (d, J=15.9 Hz , 1H), 7.36 (dd, J=2.1, 9.0 Hz, 1H), 7.32-7.19 (m, 6H), 6.60 (d, J=9.0 Hz, 1H), 6.16 (d, J=15 .9 Hz, 1H), 4.41 (q, J=6.5 Hz, 1H), 3.70 (s, 3H), 1.95-1.85 (m, 2H), 0.96 (t, J=7.4 Hz, 3H) .

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Example 5B.

To a solution of the compound of Example 5A (400 mg, 1.18 mmol) in tetrahydrofuran (5 ml) was added nickel dichloride hexahydrate (1.12 g, 4.70 mmol) and then a solution of sodium borohydride (88.92 mg, 2 .35 mmol) in dimethylformamide (1 ml), making sure that the internal temperature does not exceed 10°C, and then the reaction mixture was stirred at 10°C for 2 h, then sodium borohydride (177.84 mg, 4.70 mmol) at 0° C. and stirred at 15° C. for 2 hours. The reaction mixture was diluted with water (20 ml) and then filtered. The filtrate was extracted with ethyl acetate (20 ml x4). The combined organic layer was washed with brine (40 ml x 1), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give Example 5B, which was used directly in the next step without purification.

Example 5C.

4-(Trifluoromethoxy)phenylthioisocyanate (270.12 mg, 1.23 mmol) was added to a solution of the compound of Example 5B (350 mg, 1.12 mmol) in tetrahydrofuran (5 ml), this mixture was stirred at 25°C for 1 h Then EDCl (429.54 mg, 2.24 mmol) was added to the mixture and reacted at 70° C. for 2 hours. The reaction mixture was concentrated under reduced pressure to remove tetrahydrofuran. The residue was diluted with water (10 ml) and extracted with ethyl acetate (10 mlx3). The combined organic layer was washed with HCl (1M, 5 mL) and brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give Example 5C, which was used directly in the next step without further purification.

Example 5

Lithium hydroxide monohydrate (126.52 mg, 3.02 mmol) was added to a solution of the compound of Example 5C (500 mg, 1.01 mmol) in water (5 mL) and the mixture was stirred at 20° C. for 16 h. adjusted to pH 6-7 with HCl (1M aqueous solution) and extracted with ethyl acetate (10 ml x4). The combined organic layer was washed with brine (20 ml x 1), dried using anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue which was separated and purified by high performance liquid chromatography (TFA conditions) to give the compound of example 5. ¹ H NMR (400 MHz, methanol ₄ ) δ=67-7.60 (m, 2H), 7.56-7.43 (m, 6H), 7.42-7.36 (m, 1H), 7.32 (s, 1H), 7.08 (d, J=1.0 Hz, 2H), 5.84 (dd, J=5.5, 10.1 Hz, 1H), 2.98 (t, J=7.5 Hz, 2H), 2.76-2.65 (m, 1H), 2.65-2.54 (m,

3H), 1.06 (t, J=7.3 Hz, 3H).

Example 6

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Example 6A.

(1R)-Tetrahydronaphthalene-1 -amine (523.04 mg, 3.55 mmol) and potassium carbonate (982.04 mg, 7.11 mmol), this mixture was stirred at 50° C. for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with 100 ml of water and extracted with 200 ml (100 ml x 2) of ethyl acetate. The combined organic layer was washed with 200 ml (100 ml x 2) of brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give Example 6A, which was used directly in the next step without purification.

Example 6B.

'''ABOUT

To a solution of the compound of Example 6A (1.31 g, 3.72 mmol) in methanol (10 ml) was added nickel chloride (4.42 g, 18.59 mmol) and then a solution of sodium borohydride (1.97 g, 52.05 mmol) dissolved in dimethylformamide (3 ml), making sure that the internal temperature did not exceed 10°C, and then the reaction mixture was stirred at 10°C for 16 hours. The mixture was poured into water (100 ml), ethyl acetate ( 40 ml) and filtered through a celite filter. The filter cake was washed with ethyl acetate (60 mlx3), the mixture was extracted with 150 ml of ethyl acetate. The combined organic phase was washed with saturated brine (100 mlx2), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give Example 6B, which was used directly in the next step without purification.

Example 6C.

Isothiocyanoyl-4-(trifluoromethoxy)benzene (720.97 mg, 3.29 mmol) was added to a solution of the compound of Example 6B (1.09 g, 2.99 mmol) in tetrahydrofuran (15 ml), the mixture was stirred at 50°C. over 1 h. Then EDCl (573.25 mg, 2.99 mmol) was added and the mixture was stirred at 70° C. for 15 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with 100 ml water and extracted with 200 ml (100 mlx2) ethyl acetate. The combined organic layer was washed with 200 ml (100 mlx2) of brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0 to 12:1) to give the compound of Example 6C.

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Example 6

To a solution of the compound of Example 6C (1.36 g, 2.64 mmol) in tetrahydrofuran (10 mL) and water (4 mL) was added lithium hydroxide (554.44 mg, 13.21 mmol) and the mixture was stirred at 25° C. for for 16 hours. The mixture was acidified to pH=6 with dilute hydrochloric acid (1N, 4 ml) and extracted with 200 ml (100 mlx2) of ethyl acetate. The combined organic layer was washed with 200 ml (100 mlx2) brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was dissolved in EA (10 ml) and stirred at 25°C for 30 min, then the mixture was filtered and washed with ethyl acetate (2x10 ml) to give the compound of example 6. 1H NMR (400 MHz, methanol^ ₄ ) δ=7.60 (br d, J=8.6 Hz, 2H), 7.33-7.15 (m, 5H), 7.04 (br t, J=7.0 Hz, 1H), 6.83 (br d, J=7.6 Hz, 1H) , 6.74 (br d, J=7.9 Hz, 1H), 6.41 (br d, J=8.1 Hz, 1H), 5.88 (brt, J=8.1 Hz, 1H), 3.15-3.01 (m , 1H), 3.00-2.85 (m, 3H), 2.57 (br t, J=7.4 Hz, 2H), 2.30 (br s, 2H), 2.13 (br s, 1H), 2.01 (br d, J =16.1 Hz, 1H).

Example 7

Example 7A.

To a solution of methyl-(E)-3-(4-fluoro-3-nitrophenyl)acrylate (0.5 g, 2.22 mmol) and (R)-1-(3-chlorophenyl)ethylamine (380.12 mg, 2.44 mmol) in tetrahydrofuran (10 ml) was added potassium carbonate (613.78 mg, 4.44 mmol) and the mixed solution was stirred at 50° C. for 12 hours. The reaction solution was diluted with 30 ml of water and then extracted with ethyl acetate ( 30 mlx3). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography to give Example 7A as a yellow oil. 1H NMR (400 MHz, CDCl ₃ ) δ=8.49 (br d, J=5.5 Hz, 1H), 8.26 (d, J=2.1 Hz, 1H), 7.47 (d, J=15.9 Hz , 1H), 7.40 (dd, J=2.0, 9.0 Hz, 1H), 7.24 (s, 1H), 7.21-7.16 (m, 2H), 7.16-7.12 (m, 1H), 6.55 (d, J =9.0 Hz, 1H), 6.19 (d, J=15.9 Hz, 1H), 4.62 (quin, J=6.5 Hz, 1H), 3.71 (s, 3H), 1.59 (d, J= 6.7 Hz, 3H).

Example 7B.

Under the protection of nitrogen gas, nickel chloride (1.42 g, 10.95 mmol) was added to a solution of the compound of Example 7A (0.79 g, 2.19 mmol) in methanol (10 ml) and after cooling to 0°C, the borohydride was added slowly sodium (828.40 mg, 21.90 mmol), the mixed solution was stirred at 15° C. for 0.5 h. The reaction mixture was quenched by adding 50 ml of saturated ammonium chloride solution and stirred for 15 min. The reaction solution was extracted with ethyl acetate (50 mlx3), the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give Example 7B as a yellow oil, which was directly used in the next step without further purification.

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Example 7C.

To a solution of the compound of Example 7B (0.615 g, 1.85 mmol) in tetrahydrofuran (10 mL) was added 4(trifluoromethoxy)phenylisothiocyanate (486.02 mg, 2.22 mmol, 360.01 µm) and the mixture was stirred at 40°C for 0.5 hour, then EDC-HCl (708.46 mg, 3.70 mmol) was added and stirred at 70°C for 5 hours. The reaction solution was diluted with 30 ml of water and then extracted with ethyl acetate (30 mlx3 ). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product which was purified by column chromatography to give Example 7C as a yellow solid.

Example 7

Lithium hydroxide (234.96 mg, 5.60 mmol) was added to a mixed solution of the compound of Example 7C (0.58 g, 1.12 mmol) in tetrahydrofuran (5 ml) and water (5 ml) and stirred at 20°C in for 16 hours. The reaction solution was adjusted to pH 6 with 1M hydrochloric acid, then diluted with 30 ml of water and extracted with ethyl acetate (30 mlx3). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by high performance liquid chromatography to give the compound of example 7. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=9.27 (s, 1H), 7.93 (d, J=9.0 Hz, 2H), 7.44-7.32 (m, 5H), 7.28-7.21 (m, 2H), 6.79-6.68 (m, 2H), 6.07 (q, J=6.9 Hz, 1H), 2.82 (br t, J=7.5 Hz, 2H), 2.53 (br s, 2H), 1.90 (d, J=7.0 Hz, 3H).

Example 8

Example 8A.

K ₂ CO ₃ (920.67 mg, 6.66 mmol) and (1R)-indan-1-amine (325.33 mg, 2.44 mmol, 312.81 µl). This mixture was stirred at 50° C. for 16 hours. The reaction mixture was diluted with water (20 ml) and extracted with ethyl acetate (20 mlx3). The combined organic layer was washed with brine (20 ml x 2), dried over sodium sulfate, filtered and concentrated under reduced pressure to give

- 22 040879 compounds of example 8A. ¹ H NMR (400 MHz, CDCl ₃ ) δ=8.46 (br d, J=7.0 Hz, 1H), 8.29 (d, J=2.1 Hz, 1H), 7.60 (dd, J=2.1.9 .0 Hz, 1H), 7.54 (d, J=15.9 Hz, 1H), 7.31-7.19 (m, 4H), 7.18-7.11 (m, 1H), 7.05 (d, J=9.0 Hz, 1H),

6.26 (d, J=15.9 Hz, 1H), 5.15 (q, J=7.1 Hz, 1H), 3.73 (s, 3H), 3.08-2.99 (m, 1H), 2.96-2.86 (m, 1H), 2.70-2.60 (m, 1H), 2.02-1.96 (m, 1H).

Example 8B.

NiCl ₂ 6H ₂ O (2.02 g, 8.51 mmol) and NaBH ₄ (563.54 mg, 14.90 mmol) in portions. This mixture was stirred at 0° C. for 1 hour. The reaction mixture was quenched by adding water (50 ml) and then diluted with ethyl acetate (50 ml). The mixture was filtered and extracted with ethyl acetate (30 mlx3). The combined organic layer was washed with brine (100 ml x 1), dried over sodium sulfate, filtered and concentrated under reduced pressure to give Example 8B as a brown oil. The residue was directly used in the next step without further purification.

Example 8C.

To a solution of the compound of Example 8B (800 mg, 2.58 mmol) in THF (10 mL) was added 4(trifluoromethoxy)phenylisothiocyanate (677.91 mg, 3.09 mmol, 502.16 μL). This mixture was stirred at 30° C. for 1 hour. EDCl (988.19 mg, 5.15 mmol) was then added and the mixture was stirred at 70° C. for 2 hours. The reaction mixture was diluted with water (30 ml) and extracted with ethyl acetate (20 mlx3). The combined organic layer was washed with brine (30 ml x 1), dried over sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography to give Example 8C as a brown oil. 1H NMR (400 MHz, CDCl ₃ ) δ=7.52-7.42 (m, 3H), 7.37-7.29 (m, 2H), 7.28-7.18 (m, 3H), 7.11 (br d, J=8.7 Hz, 2Н), 7.03-6.93 (m, 1H), 6.06 (t, J=8.3 Hz, 1H), 3.70 (s, 3H), 3.30-3.20 (m, 1H), 3.16-3.09 (m, 1H) , 3.06 (t, J=7.8 Hz, 2H), 2.82-2.73 (m, 1H), 2.70 (t, J=7.9 Hz, 2H), 2.34 (qd, J=8.8, 13.4 Hz , 1H).

Example 8

To a solution of the compound of example 8C (450 mg, 908.19 µmol) in THF (2 ml) and H ₂ O (2 ml) was added LiOH-H ₂ O (114.33 mg, 2.72 mmol). This mixture was stirred at 20° C. for 16 hours. The mixture was adjusted to pH 7 with aqueous HCl (1M) and then extracted with ethyl acetate (10 mlx3). The combined organic layer was washed with brine (20 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure, and purified by high performance liquid chromatography to give the compound of example 8. 1H NMR (400 MHz, methanol-d ₄ ) δ=7.73- 7.59 (m, 2H), 7.56-7.47 (m, 3H), 7.46-7.40 (m, 1H), 7.32-7.20 (m, 3H), 6.96 (d, J=8.3 Hz, 1H), 6.41- 6.22 (m, 2H), 3.40-3.34 (m, 1H), 3.20 (td, J=8.5, 16.6 Hz, 1H), 2.99-2.93 (m, 2H), 2.92-2.84 (m, 1H), 2.62-2.55 (m, 2H), 2.55-2.46 (m, 1H).

- 23 040879

Example 9

Example 9A.

To a solution of methyl-(E)-3-(4-fluoro-3-nitrophenyl)acrylate (0.5 g, 2.22 mmol) and diphenylmethylamine (447.60 mg, 2.44 mmol, 422.26 µl) in tetrahydrofuran (10 ml) was added potassium carbonate (613.78 mg, 4.44 mmol) and the mixed solution was stirred at 50°C for 12 hours. μl), and the solution was stirred at 55°C for 12 hours. The reaction solution was diluted with 30 ml of water and then extracted with ethyl acetate (30 mlx3). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography to give Example 9A as a yellow solid. 1H NMR (400 MHz, CDCl3) δ=8.74 (br d, J=5.3 Hz, 1H), 8.27 (d, J=1.9 Hz, 1H), 7.48 (d, J=15.9 Hz, 1H), 7.42 (dd, J=1.8, 8.9 Hz, 1H), 7.32-7.20 (m, 10H), 6.69 (d, J=9.0 Hz, 1H), 6.19 (d, J=15, 9 Hz, 1H), 5.70 (d, J=5.5 Hz, 1H), 3.71 (s, 3H).

Example 9B.

Under the protection of nitrogen gas, nickel chloride (1.10 g, 8.50 mmol) was added to a solution of the compound of Example 9A (0.66 g, 1.70 mmol) in methanol (10 ml) and after cooling to 0°C, the borohydride was added slowly sodium (642.86 mg, 16.99 mmol). The mixed solution was stirred at 20° C. for 0.5 h. The reaction mixture was quenched by adding 50 ml of saturated ammonium chloride solution and stirred for 15 min. The reaction solution was extracted with ethyl acetate (50 mlx3), the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give Example 9B as a yellow oil, which was directly used in the next step without purification.

- 24 040879

Example 9C.

To a solution of the compound of Example 9B (0.41 g, 1.14 mmol) in tetrahydrofuran (10 ml) was added 4(trifluoromethoxy)phenyl isothiocyanate (278 mg, 1.27 mmol, 205.93 μl) and stirred at 40°C over 1 hour. EDC-HCl (436.11 mg, 2.27 mmol) was then added and stirred at 70°C for 12 hours. The reaction solution was diluted with 30 ml of water and then extracted with ethyl acetate (30 mlx3). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product which was purified by column chromatography to give Example 9C as a yellow solid.

Example 9

To a solution of the compound of Example 9C (0.46 g, 843.18 µmol) in tetrahydrofuran (5 ml) and water (5 ml) was added lithium hydroxide monohydrate (176.92 mg, 4.22 mmol) and stirred at 20° C. for within 12 hours

The reaction solution was adjusted to pH 6 with 1M hydrochloric acid and extracted with ethyl acetate (20 mlx3). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude product was diluted with 10 ml of petroleum ether:ethyl acetate=10:1, stirred for 30 min, filtered and concentrated under reduced pressure to give the compound of example 9. 1H NMR (400 MHz,

DMSO-d6) δ=12.06 (br s, 1H), 9.33 (br s, 1H), 7.90 (br d, J=8.2 Hz, 2H), 7.46-7.35 (m, 7H), 7.32 (br d , J=8.7 Hz, 2H), 7.26 (br s, 1H), 7.20 (br d, J=7.1 Hz, 4H), 6.64 (br d, J=7.8 Hz, 1H), 6.28 (br d, J=8.2 Hz, 1H), 2.81 (br t, J=7.5 Hz, 2H), 2.53 (brs, 2H).

Example 10

Example 10A.

Potassium carbonate (920 .67 mg, 6.66 mmol) and (1R)-1-(2-methoxyphenyl)ethylamine (503.52 mg, 3.33 mmol), this mixture was stirred at 50° C. for 16 hours. The reaction mixture was concentrated at reduced pressure to remove the solvent. The residue was diluted with 100 ml water and extracted with 200 ml (100 mlx2) ethyl acetate. The combined organic layer was washed with 200 ml (100 mlx2) of brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue, and this residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0 to 15 :1) to obtain the compound of example 10A.

Example 10B.

To a solution of the compound of Example 10A (1.19 g, 3.34 mmol) in methanol (10 ml) was added nickel chloride (3.97 g, 16.70 mmol) and then a solution of sodium borohydride (1.39 g, 36.73 mmol) in dimethylformamide (5 ml), making sure that the internal temperature does not exceed 10°C, and then the reaction mixture was stirred at 10°C for 16 hours. The mixture was poured into water (100 ml) and ethyl acetate (40 ml ) and filtered through a celite filter. The filter cake was washed with ethyl acetate (60 mlx3), the filtrate was extracted with 150 ml of ethyl acetate. The combined organic phases were washed with saturated brine (100 ml x 2), dried over anhydrous sodium sulfate and concentrated in vacuo to give Example 10B.

Example 10C.

To a solution of the compound of Example 10B (1.11 g, 3.35 mmol) in tetrahydrofuran (15 mL) was added 1isothiocyanoyl-4-(trifluoromethoxy)benzene (734.27 mg, 3.35 mmol, 543.90 μL), this mixture stirred at 50°C for 1 hour, then EDCl (642.11 mg, 3.35 mmol) was added and the mixture was stirred at 70°C for 15 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with 100 ml water and extracted with 200 ml (100 mlx2) ethyl acetate. The combined organic layer was washed with 200 ml (100 mlx2) brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0 to 6/1) to give the compound of Example 10C.

Example 10

To a solution of the compound of Example 10C (1.32 g, 2.49 mmol) in tetrahydrofuran (6 ml) and water (3 ml) was added lithium hydroxide monohydrate (523.12 mg, 12.47 mmol) and the mixture was stirred at 25 °C for 16 hours. The mixture was acidified with dilute hydrochloric acid (1N, 4 ml) to pH=6 and extracted with 200 ml (100 mlx2) of ethyl acetate. The combined organic layer was washed with 200 ml (100 mlx2) brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was dissolved in a mixed solution of ethyl acetate (4 ml) and

- 26 040879 petroleum ether (8 ml) and stirred at 25°C for 60 minutes. The mixture was filtered and the filter cake was washed with a mixed solution of ethyl acetate (1 ml) and petroleum ether (2 ml) to give the compound of example 10. ¹ H NMR (400 MHz, CDCl ₃ ) δ=7.60-7.55 (m, 2H), 7.37 -7.32(m, 1H), 7.28-7.22(m,

3H), 7.09-7.03 (m, 3H), 6.98 (dd, J=1.5, 8.3 Hz, 1H), 6.86 (d, J=8.3 Hz, 1H), 5.78 (q, J=7, 1 Hz, 1H), 3.70 (s,

3H), 3.13-2.98 (m, 2H), 2.75-2.61 (m, 2H), 1.88 (d, J=7.1 Hz, 3H).

Example 11.

Example 11A.

Potassium carbonate (675.15 mg, 2.931 mmol) and (1R)-2,2-dimethyl-1-phenyl-prop-1amine hydrochloride (195.13 mg, 977.03 µmol). This mixture was stirred at 60° C. for 48 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with 100 ml water and extracted with 200 ml (100 mlx2) ethyl acetate. The combined organic layer was washed with 200 ml (100 mlx2) brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0 to 12/1) to give the compound of Example 11A.

Example 11B.

To a solution of the compound of Example 11A (420 mg, 1.14 mmol) in methanol (6 ml) was added nickel chloride (1.35 g, 5.70 mmol) and then a solution of sodium borohydride (474.42 mg, 12 54 mmol) in dimethylformamide (3 ml), making sure that the internal temperature does not exceed 10°C, and then the reaction mixture was stirred at 10°C for 2 hours. The mixture was poured into water (80 ml), ethyl acetate (20 ml) and filtered through a celite filter. The filter cake was washed with ethyl acetate (50 mlx3) and the mixture was extracted with 100 ml of ethyl acetate. The combined organic phase was washed with saturated brine (80 ml x 2), dried over anhydrous sodium sulfate and concentrated in vacuo to give Example 11B, which was used directly in the next step without purification.

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Example 11C.

To a solution of the compound of Example 11B (270 mg, 793.05 µmol) in tetrahydrofuran (5 ml) was added 1isothiocyanato-4-(trifluoromethoxy)benzene (173.82 mg, 793.05 µmol, 128.76 µl), this mixture was stirred at 50°C for 1 hour, then EDCl (167.23 mg, 872.35 µmol) was added and the mixture was stirred at 70°C for 15 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with 100 ml water and extracted with 200 ml ethyl acetate (100 mlx2). The combined organic layer was washed with 200 ml brine (100 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue which was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate = 1/0 to 10 /1) to obtain the compound of example 11C.

Example 11.

Lithium hydroxide (145.93 mg, 3.48 mmol) was added to a solution of the compound of Example 11C (373 mg, 695.52 µmol) in tetrahydrofuran (3 ml) and water (1 ml), this mixture was stirred at 25° C. for 6 h. The mixture was acidified pH=6 with dilute hydrochloric acid (1N, 3 ml), filtered, the filter cake was washed with 20 ml of ethyl acetate to obtain a residue. The filter cake was dissolved in methanol (10 ml) and dichloromethane (10 ml), the mixture was stirred at 80°C for 1 h and then filtered. The filter cake was the product of Example 11. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=9.17 (s, 1H), 7.96 (br d, J=9.0 Hz, 2H), 7.62 (br d, J=7.6 Hz, 2H), 7.41-7.33 (m, 4H), 7.31-7.25 (m, 1H), 7.18 (s, 1H), 6.76 (d, J=8.3 Hz, 1H), 6.63 (dd, J=1.4, 8.4 Hz, 1H), 5.86 (s, 1H), 2.78 (br t, J=7.6 Hz, 2H), 2.47-2.52(m, 2H), 1.22 (s, 9H).

Example 12.

Example 12A.

Methyl (E)-3-(4-fluoro-3-nitro-phenyl)prop-2-enoate (0.26 g, 1.15 mmol) was dissolved in acetonitrile (10 ml), then potassium carbonate (558, 55 mg, 3.45 mmol) and (R)-cyclopropyl(phenyl)methylamine hydrochloride (233.30 mg, 1.27 mmol) and stirred at 80°C for 2 hours. The reaction mixture was diluted with 100 ml of water and extracted 100 ml (50 mlx2) ethyl acetate. The combined organic layer was washed with 50 ml (50 ml x 1) brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by column chromatography to give Example 12 as a yellow oil. 1H NMR (400 MHz, CDCl ₃ ) δ=0.34-0.56 (m, 2H), 0.60-0.80 (m, 2H), 1.30-1.43 (m, 1H),

- 28 040879

3.78 (s, 3H), 4.04 (dd, J=7.95, 5.38 Hz, 1H), 6.23 (d, J=15.89 Hz, 1H), 6.56 (d, J=8.93 Hz, 1H), 6.52-6.62 (m ,

1H), 7.27-7.42 (m, 6H), 7.53 (d, J=15.89 Hz, 1H), 8.32 (d, J=2.08 Hz, 1H), 8.88 (br d, J=5.01 Hz, 1H).

Example 12V.

To Example 12A (388 mg, 1.10 mmol) and nickel chloride hexahydrate (1.31 g, 5.51 mmol) were added MeOH (6 ml) and THF (6 ml) and sodium borohydride (416.54 mg, 11.01 mmol) at 5° C. and stirred at 5° C. for 0.5 h. The reaction mixture was quenched by adding 100 ml of water, filtered, and the filtrate was extracted with 100 ml (50 ml x 2) of ethyl acetate. The combined organic layer was washed with 50 ml (50 ml x 1) brine, dried over sodium sulfate, filtered and concentrated under reduced pressure to give Example 12B.

Example 12C.

O-CF ₃

To a solution of the compound of Example 12B (280 mg, 863.09 µmol) in THF (15 ml) was added isothiocyanoyl-4-(trifluoromethoxy)benzene (227.01 mg, 1.04 mmol, 168.16 µl). After the addition, the mixture was stirred at 40° C. for 1 h and then EDCl (330.91 mg, 1.73 mmol) was added and stirred at 70° C. for 1 h. The reaction mixture was concentrated under reduced pressure to remove THF, diluted with 80 ml of water and extracted with 100 ml (50 mlx2) of ethyl acetate. The combined organic layer was washed with 50 ml (50 ml x 1) brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by column chromatography to give Example 12C as a colorless oil. 1Н NMR (400 MHz, CDCl ₃ ) δ=0.47-0.70 (m, 3Н), 0.95-1.07 (m, 1Н), 1.74-1.83 (m, 1Н), 2.70 (t, J=7.89 Hz, 2Н), 3.06 (t, J=7.95 Hz, 2H), 3.69 (s, 3H), 4.84 (d, J=9.54 Hz, 1H), 6.97 (dd, J=8.19, 1.34 Hz, 1H), 7.05-7.16 (m , 3H), 7.28-7.33 (m, 2H), 7.40-7.54 (m, 6H).

Example 12.

To a solution of Example 12C (0.29 g, 569.16 µmol) in THF (3 ml), MeOH (3 ml) and H2O (3 ml) was added lithium hydroxide monohydrate (119.41 mg, 2.85 mmol) and stirred at 20° C. for 16 hours. The reaction mixture was slowly adjusted to pH=4 with 1N. HCl solution and extracted with ethyl acetate (40 mlx2). The combined organic layer was washed with 50 ml of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, stirred with MeOH (10 ml), filtered and dried under vacuum to give the compound of example 12. 1H NMR (400 MHz, CDCl ₃ ) δ= 4.38 (br s, 1Н), 4.79-4.97 (m, 1Н), 6.51-6.62 (m, 2Н), 7.25-7.27 (m, 1Н), 7.27-7.29 (m, 1Н), 7.38-7.51 (m, 5H).

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Example 13

Example 13A.

3,3-Dimethylinden-1-one (3 g, 18.73 mmol) was dissolved in toluene (40 ml), titanium tetraisopropoxide (10.64 g, 37.46 mmol, 11.05 ml) was added thereto and 2- methylpropane-2-sulfinamide (2.72 g, 22.48 mmol) and stirred at 60°C for 24 h, then again added 2-methylpropane-2-sulfinamide (2.72 g, 22.48 mmol) and tetraisopropoxide titanium (5.32 g, 18.73 mmol, 5.53 ml) and stirred at 110° C. for 16 hours. The reaction mixture was quenched by adding 200 ml of water and then filtered. The filtrate was extracted with 160 ml (80 ml x 2) ethyl acetate. The combined organic layer was washed with 80 ml (80 ml x 1) brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by column chromatography to give Example 13A as a brown oil. 1H NMR (400 MHz, CDCl ₃ ) δ=1.29-1.43 (m, 15 H) 2.98 (d, J=19.07 Hz, 1H) 3.34 (d, J=19.20 Hz, 1H) 7.29-7.42 (m, 2H) 7.48-7.58 (m, 1H) 7.76 (d, J=7.70 Hz, 1H).

Example 13B.

The compound of example 13A (1.4 g, 5.32 mmol) was dissolved in THF (20 ml) and water (574.67 mg, 31.89 mmol, 574.67 μl), sodium borohydride (1.01 g , 26.58 mmol) and this mixture was stirred at 25° C. for 4 hours. The reaction mixture was quenched by adding 100 ml of water and then extracted with 100 ml (50 ml x 2) of ethyl acetate.

The combined organic layer was washed with 50 ml brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by column chromatography to give Example 13B as a white solid. Ή NMR (400 MHz, CDCl ₃ ) δ=1.21-1.28 (m, 12Н) 1.38 (s, 3Н) 1.88 (dd, J=12.84, 7.46 Hz, 1Н) 2.37 (dd, J=12.84, 7.21 Hz, 1Н ) 3.49 (br d, J=6.60 Hz, 1Н) 4.88-4.99 (m, 1Н) 7.14-7.21 (m, 1Н) 7.23-7.29 (m, 2Н) 7.51-7.60 (m, 1Н).

Example 13C. (1R)-3,3-Dimethylinden-1-amine.

To a solution of the compound of example 13B (810 mg, 3.05 mmol) in MeOH (8 ml) was added HCl/dioxane (4M, 3 ml) and stirred at 20°C for 0.5 h. The mixture was diluted with methyl tert-butyl ether (15 ml) and stirred at 25°C for 30 min, filtered and dried in vacuum to obtain the compound of example 13C as a white solid. 1H NMR (400 MHz, methanol-d ₄ ) δ=1.15-1.30 (m, 3H) 1.40-1.50 (m, 3H) 1.93 (dd, J=13.02, 8.13 Hz, 1H) 2.53 (dd, J=12.96, 7.70 Hz, 1H) 3.31 (dt, J=3.30, 1.65 Hz, 1H) 7.21-7.59 (m, 4H).

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Example 13D.

Potassium carbonate (982.04 mg, 7.11 mmol) and the compound of example 13C (491.69 mg, 2.49 mmol) and stirred at 65°C for 16 hours. The mixture was diluted with 100 ml of water and extracted with 120 ml (60 ml x 2) of ethyl acetate. The combined organic layer was washed with 50 ml (50 ml x 1) brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by column chromatography to give Example 13D as a yellow oil. 1Н NMR (400 MHz, CDCl ₃ ) δ=1.33 (s, 3Н) 1.43 (s, 3Н) 1.95 (dd, J=12.72, 7.34 Hz, 1Н) 2.60 (dd, J=12.65, 7.15 Hz, 1Н) 3.82 (s, 3H) 5.25 (q, J=7.21 Hz, 1H) 6.34 (d, J=15.89 Hz, 1H) 7.14 (d, J=9.05 Hz, 1H) 7.25-7.27 (m, 1H) 7.27-7.29 ( m, 1H) 7.30-7.39 (m, 2H) 7.57-7.73 (m, 2H) 8.38 (d, J=2.20 Hz, 1H) 8.57 (br d, J=7.09 Hz, 1H).

Example 13E.

Example 13D (300 mg, 818.75 µmol) was dissolved in THF (5 ml) and MeOH (5 ml), Pd/C (50 mg, 10%) was added thereto, the atmosphere was replaced with hydrogen gas 3 times, and the mixture was stirred at 25°C in an atmosphere of H ₂ (15 psi) for 16 hours After filtration, the filtrate was concentrated under reduced pressure to obtain the compound of example 13E as a brown oil.

Example 13F.

Example 13E (255 mg, 753.45 µmol) was dissolved in THF (8 ml), 1isothiocyanoyl-4-(trifluoromethoxy)benzene (198.17 mg, 904.14 µmol, 146.79 µl) was added thereto and stirred at 40°C for 1 h, then EDCl (288.87 mg, 1.51 mmol) was added and stirred at 70°C for 1 h. The reaction mixture was concentrated under reduced pressure to remove THF, diluted with 80 ml of water and extracted with 100 ml (50 mlx2) ethyl acetate. The combined organic layer was washed with 50 ml (50 ml x 1) brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by column chromatography, and then subjected to chiral separation to give Example 13F as a white solid. 1H NMR (400 MHz, CDCl ₃ ) δ=1.28-1.55 (m, 6Н) 2.14-2.72 (m, 4Н) 3.06 (br s, 2Н) 3.69 (s, 3Н) 5.50 (br s, 1Н) 6.06 ( br s, 1H) 6.92–7.60 (m, 11H).

Example 13

To a solution of Example 13F (180 mg, 343.81 µmol) in MeOH (2M1) was added monohydrate

- 31 040879 lithium hydroxide (43.28 mg, 1.03 mmol), THF (2 ml) and H ₂ O (2 ml) and stirred at 25°C for

h. The reaction mixture was slowly adjusted to pH=4 1 n. HCl, and then the precipitate was collected by filtration and dried under vacuum to obtain the compound of example 13. 1H NMR (400 MHz, methanol-d ₄ ) δ=1.34 (s,

3Н) 1.54 (s, 3Н) 2.37 (dd, J=12.65, 9.48 Hz, 1Н) 2.48-2.63 (m, 3Н) 2.90 (br t, J=7.58 Hz, 2Н) 6.05-6.37 (m, 2Н) 6.69 (br d, J=8.07 Hz, 1H) 6.93 (br d, J=7.58 Hz, 1H) 7.14-7.32 (m, 4 H) 7.35-7.42 (m, 2H) 7.59 (br d, J=7.58 Hz, 2H).

Example 14 and Example 15.

Example 14A.

A dry vessel was charged with 1-(trifluoromethyl)-1,2-benzoiodo-3-one (3.28 g, 10.37 mmol) and CuBr (123.96 mg, 864.15 µmol, 26.32 µl). The vessel was evacuated and filled with nitrogen gas three times. Then azido(trimethyl)silane (2.49 g, 21.60 mmol, 2.84 ml) and styrene (900 mg, 8.64 mmol) dissolved in ACN (40 ml) were added to the above vessel. The reaction mixture was stirred at 40° C. for 1.5 h. The reaction mixture was diluted with water (40 ml) and extracted with DCM (20 mlx3). The combined organic layer was washed with brine (20 ml x 1), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue which was purified by column chromatography to give Example 14A as a colorless oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ=7.50-7.36 (m, 5H), 4.81 (dd, J=5.1, 8.4 Hz, 1H), 2.73-2.45 (m, 2H).

Example 14B.

To a solution of the compound of example 14A (1.1 g, 5.11 mmol) in MeOH (15 ml) was added CuSO ₄ (81.59 mg, 511.21 μmol, 78.46 μl) and NaBH ₄ (290.11 mg, 7.67 mmol). This mixture was stirred at 0°C for 1 hour and then at 20°C for 16 hours. The reaction mixture was diluted with water (20 ml) and extracted with ethyl acetate (20 mlx3). The combined organic layer was washed with brine (20 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give Example 14B as a brown oil which was used in the next step without further purification.

Example 14C.

K2CO3 (1.10 g, 7 .99 mmol) and Example 14B (720 mg, 3.81 mmol). This mixture was stirred at 50° C. for 16 hours. The mixture was then stirred at 70° C. for 32 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with water (50 ml) and extracted with ethyl acetate (50 mlx3). The combined organic layer was washed with brine (50 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product of Example 14C as a yellow oil which was used in the next step without further purification.

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Example 14D.

NiCl ₂ -6H ₂ O (2.41 g, 10.14 mmol) was added to a solution of the compound of example 14C (1 g, 2.54 mmol) in MeOH (20 ml), then NaBH ₄ (959.38 mg, 25.36 mmol) at 0°C and this mixture was kept at 0°C for 1 hour. The reaction mixture was diluted with water (50 ml) and extracted with ethyl acetate (50 mlx3). The combined organic layer was washed with brine (50 ml x 1), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give Example 14D as a brown oil which was used directly in the next step without further purification.

Example 14E and Example 14F.

Example 14E or Example 14F Example 14E or Example 14F

To Example 14D (770 mg, 2.10 mmol) in THF (20 ml) was added 1-isothiocyanoyl-4(trifluoromethoxy)benzene (552.78 mg, 2.52 mmol, 409.47 μl). This mixture was stirred at 40° C. for 1 h. EDCl (805.79 mg, 4.20 mmol) was then added and the mixture was stirred at 70° C. for 2 h. The reaction mixture was diluted with water (50 ml) and extracted with ethyl acetate (30 mlx3). The combined organic layer was washed with brine (50 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue which was purified by column chromatography to give a residue. The residue was separated by SFC (supercritical fluid chromatography) (chiral column: Chiralcel OD-3 50 x 4.6 mm id, 3 μm; mobile phase: A: CO ₂ , B: methanol (containing 0.05% DEA); gradient : methanol from 5% to 40%; flow rate: 3 ml/min; wavelength: 220 nm) to give the compound of example 14E (retention time RT=1.546 min) and the compound of example 14F (retention time RT=1.941 min).

Example 14 and Example 15.

Example 14 or Example 15 Example 14 or Example 15

To a solution of Example 14E (220 mg, 39.93 µmol) in MeOH (3 ml) and H2O (3 ml) was added LiOH-H ₂ O (50.22 mg, 1.20 mmol). This mixture was stirred at 30°C for 16 hours. The mixture was stirred at 30°C for another 3 hours. The mixture was adjusted to pH 5-6 with HCl solution (1M) to precipitate a solid, which was filtered and dried to give the compound of example 14 ?Н NMR (400 MHz, methanol-d ₄ ) δ=7.57-7.47 (m, 2Н), 7.43 (d, J=4.4 Hz, 4Н), 7.38-7.33 (m, 1H), 7.30 (s, 1H), 7.25 (d, J=8.3Hz, 2H), 6.88 (d, J=0.6Hz, 2H), 6.30 (dd, J=3.6, 10.5Hz, 1H), 3.61-3.37 (m, 2H), 2.96 (t, J=7.7 Hz, 2H), 2.61 (t, J=7.7 Hz, 2H).

To a solution of Example 14F (300 mg, 543.99 µmol) in MeOH (4 ml) and H2O (4 ml) was added LiOH-H ₂ O (68.48 mg, 1.63 mmol). This mixture was stirred at 30° C. for 16 hours. The mixture was brought to

- 33 040879 pH 5-6 HCl solution (1M) to precipitate a solid, which was filtered and dried to give the compound of example 15. ¹ H NMR (400 MHz, methanol-d ₄ ) δ=7.55-7.49 (m, 2H), 7.43 (d, J=4.4 Hz,

4H), 7.38-7.33 (m, 1H), 7.30 (s, 1H), 7.28-7.21 (m, 2H), 6.87 (s, 2H), 6.30 (dd, J=3.5, 10.5 Hz, 1H) , 3.56-3.41 (m, 2H), 2.96 (t, J=7.6 Hz, 2H), 2.61 (t, J=7.7 Hz, 2H).

Experimental Example 1 In vitro IDH1 activity test.

Mutant IDH1 catalyses the NADPH-dependent reduction of α-KG (α-ketoglutarate) to 2-HG (2-hydroxyglutarate) and NADPH consumed can be determined by fluorescence reading.

Reagents.

Basic reaction buffer: 50 mM KH2PO4, pH 7.5, 10 mM MgCl ₂ , 10% glycerol, 150 mM NaCl, 0.05% BSA (bovine serum albumin), 2 mM b-ME (2-mercaptoethanol), 0.003% Brij35 (polyethylene glycol lauryl ether).

substrates and cofactors.

IDHlwt (wild type): 65 µM isocitrate + 50 µM NADP.

IDH1(R132H): 1500 µM α-KG + 15 µM NADPH.

IDH1(R132C): 500 µM α-KG + 15 µM NADPH.

reaction process.

1.33x enzyme (no enzyme in control well), buffer, and NADP or NADPH (control well) were added to the wells of the reaction plate. The test compound was dissolved in 100% DMSO and then added to the enzyme mixture (Echo550, nanoliter level) and incubated for 60 minutes after simple centrifugation. A mixture of 4x substrate and cofactor was added to start the reaction. After simple centrifugation, the plate was incubated with shaking at room temperature for 45 minutes. A mixture of 3x lipoamide hydrogenase and resazurin was prepared, added to the reaction solution to test the amount of NADPH formed or remaining, and incubated at room temperature for 10 minutes after simple centrifugation. An Envision multifunctional microplate reader was used to measure (E _x /E _m =535/590 nm).

The results of the experiments are presented in table. 1 and table. 2.

Table 1. Results of IC ₅₀ testing for IDH1 (IDH1R132H) enzyme activity in vitro

Example Structure IDH1 R132H (nM) Example 1 OCF ₃ .А™ 0 HU η \\ / | 1 u-NH 7.9 Example 2 I o \= o A o o 464.5

- 34 040879

Example 3 I o \= o O o ω 320 Example 4 OCF ₃ I JL % ^NH 125.5 Example 5 OCF ₃ ; 0 I JL % ^NH 89.53 Example 6 X Oh / Oh 3.94 Example 7 X Oh / ΥΥ 121.1 Example 8 F p-(- ^F _Η0 Α _γ . » 0 ^{F but} Т1Нн 00 2.88 Example 9 0 about 21.9

- 35 040879

Table 2. Results of IC50 testing for IDH1 enzyme activity (IDH1 R132C, WT) in vitro

Example Structure IDH1 R132C (nM) IDH1 WT (nM) Example 1 OCF ₃ ^ΗΟ ^ΏΓΧ 101.8 >10000 Example 6 OCF ₃ Λ Ο ^ηοΛ ^Ο: ^ν μ 12.79 >10000

Conclusion.

At the enzymatic level, the compounds of the present invention have a good inhibitory effect on the mutant IDH1R132H and IDH1R132C and at the same time have no inhibitory effect on the wild-type IDH protein.

Experimental Example 2 Testing the cytological activity of IDH1.

In this study, the compound was incubated with a cell line with mutant IDH1, and the content of 2HG in the cell culture was determined by LC-MS (liquid chromatography/mass spectrometry) to determine the inhibitory activity of the compound against mutant IDH1. IDH1 catalyzes the reduction of isocitrate to α-ketoglutarate (α-KG) in organisms, and mutant IDH1 additionally catalyzes the reduction of α-KG to 2-hydroxyglutarate (2-HG).

The U87MG-IDH1-R132H cell strain is a stable transfected cell strain that can stably express the IDH1-R132H mutant obtained by screening U87MG cells with the IDH1-R132H mutant, and the HT1080 cell strain contains the endogenous IDH1-R132C mutant.

The experimental process is the following.

1) The compound was subjected to 3-fold dilutions in DMSO and then added to the cell culture plate for a total of 10 concentrations, in duplicate for each concentration. The negative control well contained only DMSO and the positive control well contained BAY1436032 at a final concentration of 5 μM. The final concentration of DMSO in all wells was 0.5%.

2) The mutant IDH1 cell line was seeded in a cell culture plate containing the compound at a density of 40,000 cells per well. Cells were incubated with the compound in an incubator at 37°C, 5% CO2 for 3 days.

3) After 3 days, 10 µl of the cell culture supernatant was taken and diluted 21 times with 200 µl of ddH ₂ O water to 210 µl and mixed well, 50 µl of the diluted solution was taken from them and 200 µl of the precipitant (acetonitrile containing 0.4 µg/ml D-2-hydroxyglutaric acid ¹³ C5). After centrifugation at 4000 rpm for 10 min, 100 µl of the

- 36 040879 for the determination of 2-HG content by LC-MS.

4) At the same time, the ATPlite 1Step kit was used to determine the effect of the compound on cell viability of the IDH1 mutant cell strain according to the instructions.

5) The 2-HG content data was used to calculate the percent inhibition (degree of inhibition, %) for each compound concentration against the IDH1 mutant using the following calculation formula:

Degree of inhibition, %=(CPD-ZPE)/(HPE-ZPE)x100%

Cell viability data was used to calculate the percent cytotoxicity (cytotoxicity, %) of the compound against the IDH1 mutant cell strain using the following calculation formula:

Cytotoxicity, %=( 1 -CPD/ZPE)x 100%

CPD: signal from wells with connection.

ZPE: Average signal from negative control wells with 0.5% DMSO instead of compound.

HPE: average signal from positive control wells.

6) The degree of inhibition, %, and cytotoxicity, %, were fitted using the GraphPad Prism software to a dose-response curve to obtain the IC ₅₀ value of the test compound.

The results of the experiment are presented in table. 3.

Table 3. IC ₅₀ test results for

Conclusion.

At the cellular level, the compounds of the present invention have a good 2-HG inhibitory effect on brain U87MG glioma cells with an IDH1R132H mutation.

Experimental Example 3 Pharmacokinetic Evaluation in Mice.

Target.

To test the pharmacokinetic parameters of the compound of the present invention in mice.

Experiment scheme:

1) experimental drug: the compound of example 6;

2) experimental animals: 8 male CD-1 mice aged 7-10 weeks, divided into 2 groups, 4 in each group;

3) drug preparation: for the tail vein injection group, an appropriate amount of drug was weighed and dissolved in a mixed solvent of DMSO:20% hydroxypropyl-betacyclodextrin (HPbCD)=10:90 to prepare a solution of 0.5 mg/ml; for the intragastric group, the appropriate amount of drug was weighed and dissolved in DMSO:polyoxyethylene castor oil EL (Cremophor EL):5% sulfobutylcyclodestrin (Captisol)=5:10:85 to prepare a suspension.

Conducting an experiment.

Animals in group 1 received a single injection of the drug at a dose of 1 mg/kg and at a concentration of 0.5 mg/ml through the tail vein, and animals in group 2 were injected with the compound at a dose of 20 mg/kg and at a concentration of 2 mg/ml through the gastric probe. Plasma samples were cross-collected from animals at 0.0833 (tail vein injection group only), 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours post-administration. The LC-MS/MS method was used to determine the concentration of a drug in a blood plasma sample, and the kinetic parameters of the test drug are presented in table. 4.

-

Claims (11)

Table 4. Estimated pharmacokinetic parameters in mice Tail vein injection group Clearance Initial concentration Volume of distribution Half-life Area under the curve C1 (ml/min/kg) Co (nM) Vd (l/kg) Т1/2 (H) AUCo-last (nM*h) 2.17 3541 0.690 2.90 15463 Group of intragastric administration highest concentration Time to highest concentration Area under the curve Bioaccess- ness Stach (nM) Tsh(h) AUCo-last (nM*h) F(%) 33350 1.00 306609 99.1 Conclusion. The compound of Example 6 of the present invention has good pharmacokinetic properties in mice. CLAIM 1. The compound as it is represented in formula (I), (I) where R1 is selected from C _1-6 alkyl, C _3-6 cycloalkyl and phenyl, with C _1-6 alkyl, C _3-6 cycloalkyl and phenyl optionally substituted with 1, 2 or 3 Ra; each of R ₂ and R 3 is independently selected from H, F, Cl, Br, I, OH, NH ₂ , CN, COOH, C _1-6 alkyl and C _1-6 alkoxy, with C _1-6 alkyl and C _{1- 6} alkoxy possibly substituted with 1, 2 or 3 R _b ; alternatively R1 and R2 are linked together to form a C _4-6 cycloalkenyl, optionally substituted with 1, 2 or 3 Rc; L is -CH2CH2-; n is selected from 1, 2 and 3; and each of Ra, R _b , and Rc is independently selected from F, Cl, Br, I, OH, NH2, CN, COOH, and Me; or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. 2. The compound, or its stereoisomer, or its pharmaceutically acceptable salt according to claim 1, where R1 is selected from C _1-3 alkyl, cyclopropanyl and phenyl, with C _1-3 alkyl, cyclopropanyl and phenyl optionally substituted with 1, 2 or 3 Ra, or each of R2 and R3 is independently selected from H, F, Cl, Br , I, OH, NH2, CN, COOH, C _1-3 alkyl and C _1-3 alkoxy, with C _1-3 alkyl and C _1-3 alkoxy optionally substituted with 1, 2 or 3 R _b ; or L is -CH2CH2-; or p-· 'I-rg ² R _{3 and} ^r 2 structural unit selected from ' or - 38 040879 from 3. A compound, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 2, wherein R1 is branes from CH3, _CH2F , _CHF2 , _CF3 , CHCIK _CH2CF3 , _CH2CH2CH3 , C( _{CH3)3 ,} V, _CH (CH3) ₂ and 4. A compound, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 2, wherein R2 and _R3 are each independently selected from H, F, Cl, Br, I, OH, NH2, CN, COOH, _CH3 and CH ₃ O, with CH ₃ and CH ₃ O possibly substituted with 1, 2 or 3 Rb. 5. A compound, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 2, wherein R2 and _R3 are each independently selected from H, F, Cl, Br, I, OH, NH2, CN, COOH, _CH3 , CH2F , CHF2, CF ₃ and OCH ₃ . 6. Compound or stereoisomer or pharmaceutically acceptable salt thereof according to claim 2, where the structure according to claim 2, where the structure 7. The compound, or its stereoisomer, or its pharmaceutically acceptable salt, is the chosen unit. its pharmaceutically acceptable salt according to any one of 8. The compound, or its stereoisomer, or claims 1, 2, which is selected from among - 39 040879 Where E is selected from -CH2-, -CH2CH2-, -C(CH3) ₂ - and -C(CH3) ₂ CH2—; L represents -CH ₂ CH ₂ -; R1 is as defined in any of claims 1, 2; And R ₂ and R3 are as defined in any one of claims 1, 2. 9. A compound, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 8, which is selected from 10. A compound, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 1, which is selected from - 40 040879 or which is selected from - 41 040879 - 42 040879 11. The use of a compound, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 10 in the manufacture of a medicament for the treatment of a disease associated with IDH1 (isocitrate dehydrogenase 1). Eurasian Patent Organization, EAPO Russia, 109012, Moscow, Maly Cherkassky per., 2