Spirocyclic amine substituted pyridazine or 1,2,4-triazine

Abstract

The present invention relates to medicaments useful for therapy and / or prophylaxis in mammals, pharmaceutical compositions comprising such compounds, and their use as menin / MLL protein / protein interaction inhibitors useful in treating diseases such as cancer, including but not limited to leukemia, myelodysplastic syndromes (MDS), and myeloproliferative neoplasms (MPN), and diabetes.

Description

[Technical field]

[0001] The present invention relates to medicaments useful for therapy and / or prophylaxis in mammals, pharmaceutical compositions comprising such compounds, and their use as menin / MLL protein / protein interaction inhibitors useful in treating diseases such as cancer, including but not limited to leukemia, myelodysplastic syndrome (MDS), and myeloproliferative neoplasms (MPN), and diabetes. [Background technology]

[0002] Chromosomal rearrangements affecting the mixed lineage leukemia gene (MLL; MLL1, KMT2A) result in aggressive acute leukemia across all age groups that remains largely incurable, highlighting the urgent need for novel therapeutic approaches. Acute leukemias with these chromosomal translocations of MLL are expressed as lymphocytic, myeloid or biphenotypic diseases and constitute 5-10% of acute leukemias in adults and approximately 70% in infants (Marschalek, Br J Haematol 2011.152(2),141-54; Tomizawa et al., Pediatr Blood Cancer 2007.49(2),127-32).

[0003] MLL is a histone methyltransferase that methylates histone H3 on lysine 4 (H3K4) and functions in a multiprotein complex. The use of inducible loss-of-function alleles of Mll1 demonstrated that Mll1 plays an essential role in hematopoietic stem cell (HSC) maintenance and B cell development, but its histone methyltransferase activity is dispensable for hematopoiesis (Mishra et al., Cell Rep 2014.7(4),1239-47).

[0004] Fusions of MLL with over 60 different partners have been reported to date and are associated with leukemia formation / progression (Meyer et al., Leukemia 2013.27,2165-2176). Interestingly, the SET (Su(var)3-9, enhancer of zeste, and trithorax) domain of MLL is not retained in the chimeric protein but is replaced by the fusion partner (Thiel et al., Bioessays 2012.34,771-80). Recruitment of chromatin-modifying enzymes such as Dot1L and / or pTEFb complex by the fusion partner results in enhanced transcription and transcriptional elongation of MLL target genes, including most notably HOXA genes (e.g., HOXA9) and the HOX cofactor MEIS1. In turn, aberrant expression of these genes blocks hematopoietic differentiation and enhances proliferation.

[0005] Menin, encoded by the Multiple Endocrine Neoplasia type 1 (MEN1) gene, is ubiquitously expressed and mainly localized in the nucleus. It has been shown to interact with numerous proteins and is therefore involved in a variety of cellular processes. The best understood function of menin is its role as an oncogenic cofactor of MLL fusion proteins. Menin interacts with two motifs, MBM1 (menin-binding motif 1) and MBM2, within the N-terminal fragment of MLL that is retained in all fusion proteins (Thiel et al., Bioessays 2012.34,771-80). Menin / MLL interaction results in the formation of a new interaction surface for lens epithelium-derived growth factor (LEDGF). While MLL directly binds to LEDGF, menin is essential for stable interaction between MLL and LEDGF and gene-specific chromatin recruitment of the MLL complex via the PWWP domain of LEDGF (Cermakova et al., Cancer Res 2014.15,5139-51; Yokoyama & Cleary, Cancer Cell 2008.8,36-46). Furthermore, many genetic studies have shown that menin is strictly required for oncogenic transformation by MLL fusion proteins, suggesting the menin / MLL interaction as an attractive therapeutic target. For example, conditional deletion of Men1 prevents leukocyte formation in myeloid progenitor cells ectopically expressing MLL fusions (Chen et al., Proc Natl Acad Sci 2006.103,1018-23). Similarly, genetic disruption of the menin / MLL fusion interaction by loss-of-function mutations abolishes the oncogenic properties of MLL fusion proteins, blocks leukemia development in vivo, and releases the differentiation block of MLL-transformed leukemic blasts. These studies also showed that menin is required for the maintenance of HOX gene expression by MLL fusion proteins (Yokoyama et al., Cell 2005.123,207-18).Furthermore, small molecule inhibitors of the menin / MLL interaction have been developed, suggesting the druggability of this protein / protein interaction and demonstrating efficacy in preclinical models of AML (Borkin et al., Cancer Cell 2015.27,589-602; Cierpicki and Grembecka, Future Med Chem 2014.6,447-462). Together with the observation that menin is not an essential cofactor for MLL1 during normal hematopoiesis (Li et al., Blood 2013.122,2039-2046), these data validate disruption of the menin / MLL interaction as a promising new therapeutic approach for the treatment of MLL-rearranged leukemias and other cancers with an active HOX / MEIS1 gene signature. For example, internal partial tandem duplication (PTD) within the 5' region of the MLL gene represents another major abnormality found primarily in de novo and secondary AML and myelodysplastic syndromes. Although the molecular mechanisms and biological functions of MLL-PTD are not fully understood, new therapeutic targeting strategies affecting menin / MLL interaction may also prove effective in the treatment of MLL-PTD-associated leukemia. Furthermore, castration-resistant prostate cancer has been shown to be dependent on menin / MLL interaction (Malik et al., Nat Med 2015.21,344-52).

[0006] The MLL protein is also known in the scientific community as the histone-lysine N-methyltransferase 2A (KMT2A) protein (UniProt accession number Q03164).

[0007] Several references describe inhibitors targeting the menin-MLL interaction: WO 2011029054, J Med Chem 2016, 59, 892-913 describes the preparation of thienopyrimidine and benzodiazepine derivatives; WO 2014164543 describes thienopyrimidine and thienopyridine derivatives; Nature Chemical Biology March 2012, 8, 277-284 and Ren, J.; et al. Bioorg Med Chem Lett (2016), 26(18), 4472-4476 describe thienopyrimidine derivatives; J Med Chem 2014, 57, 1543-1556 describe hydroxy- and aminomethylpiperidine derivatives; Future Med Chem WO 2014,6,447-462 reviews small molecule and peptidomimetic compounds; WO 2016195776 reviews furo[2,3-d]pyrimidines, 9H-purines, [1,3]oxazolo[5,4-d]pyrimidines, [1,3]oxazolo[4,5-d]pyrimidines, [1,3]thiazolo[5,4-d]pyrimidines, thieno[2,3-b]pyridines and thieno[2,3-d]pyridines. Pyrimidine derivatives are described; WO2016197027 describes 5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine, 5,6,7,8-tetrahydropyrido[4,3-d]pyrimidine, pyrido[2,3-d]pyrimidine and quinoline derivatives, and WO2016040330 describes thienopyrimidine and thienopyridine compounds. WO2017192543 describes piperidines as menin inhibitors. WO2017112768, WO2017207387, WO2017214367, WO2018053267 and WO2018024602 describe inhibitors of menin-MLL interaction. WO2017161002 and WO2017161028 describe inhibitors of menin-MLL. WO2018050686, WO2018050684 and WO2018109088 describe inhibitors of menin-MLL interaction.WO2018226976 describes methods and compositions for inhibiting the interaction of menin with MLL proteins. WO2018175746 provides methods for treating hematological malignancies and Ewing's sarcoma. WO2018106818 and WO2018106820 provide methods for promoting the proliferation of pancreatic cells. WO2018153312 discloses azaspiro compounds related to the field of medicinal chemistry. WO2017132398 discloses a method comprising contacting leukemia cells exhibiting an NPM1 mutation with a pharmacological inhibitor of the interaction between MLL and menin. WO2019060365 describes substitution inhibitors of menin-MLL. WO2020069027 describes the treatment of hematological malignancies with inhibitors of menin. Krivtsov et al., Cancer Cell 2019. No. 6 Vol. 36, 660-673, describe menin-MLL inhibitors.

[0008] WO2021121327 describes substituted linear spiro derivatives and their use as menin / MLL protein / protein interaction inhibitors. Summary of the Invention [Means for solving the problem]

[0009] The present invention relates to a compound of formula (I):

[0010] [ka] [In the formula, R 1a -C(=O)-NR xa R xb , Het or

[0011] [ka] represents Het represents a 5- or 6-membered monocyclic aromatic ring containing 1, 2 or 3 nitrogen atoms and, optionally, a carbonyl moiety; The 5- or 6-membered monocyclic aromatic ring is optionally 3~6 Cycloalkyl and C 1~4 substituted with 1, 2 or 3 substituents selected from the group consisting of alkyl; R xa and R xb are each independently hydrogen; C 1~4 Alkyl;C 3~6 Cycloalkyl; C substituted with 1, 2 or 3 halo atoms 1~4 Alkyl; and one -OH, -OC 1~4 Alkyl or NR 11c R 11d C replaced with 1~4 alkyl; R 1b represents F or Cl, R 1c represents H or halo; Y 1 But -CR 5a R 5b -, -O- or -NR 5c - represents R 2 But hydrogen, halo, C 1~4 Alkyl, -OC 1~4 Alkyl, and -NR 7a R 7b is selected from the group consisting of U represents N or CH; n1, n2, n3 and n4 are each independently selected from 1 and 2; X 1 represents CH, and X 2 represents N, R 4 But, C 1~5 Alkyl,

[0012] [ka] represents R 5a , R 5b , R5c , R 7a , and R 7b are each independently hydrogen, C 1~4 Alkyl, and C 3~6 cycloalkyl; R 3 But -C 1~6 Alkyl-NR 8a R 8b , -C 1~6 Alkyl-C(=O)-NR 9a R 9b , -C 1~6 Alkyl-OH or -C 1~6 Alkyl-NR 11 -C(=O)-OC 1~4 Alkyl-OC(=O)-C 1~4 represents an alkyl group, R 3 C in the definition of 1~4 Alkyl or C 1~6 Each of the alkyl moieties, independently of the others, is selected from cyano, halo, -OH, and -OC 1~4 and optionally substituted with 1, 2, or 3 substituents each independently selected from the group consisting of alkyl, R 8a and R 8b are each independently hydrogen; C 1~6 Alkyl; -C(=O)-C 1~4 Alkyl; -C(=O)-OC 1~4 Alkyl; -C(=O)-NR 12a R 12b ; and -OH, cyano, halo, -C≡C, -CH=CH, -SC 1~4 Alkyl, -S(=O)2-C 1~4 Alkyl, -S(=O)2-NR 11a R 11b , -OC 1~4 Alkyl, -C(=O)-NR 10a R 10b , -NR 10c -C(=O)-C 1~4 Alkyl, 1 NR 11a R 11b -OC is substituted with 1~4 -OC substituted with alkyl and 1, 2 or 3 halo atoms1~4 C substituted with 1, 2 or 3 substituents each independently selected from the group consisting of alkyl 1~6 alkyl; R 9a , R 9b , R 10a , R 10b , R 10c , R 11 , R 11a , R 11b , R 12a , and R 12b each independently represents hydrogen and C 1~6 is selected from the group consisting of alkyl, R 11c and R 11d each independently represents hydrogen, C 1~6 Alkyl and -C(=O)-C 1~4 and tautomeric and stereoisomeric forms thereof; and pharma- ceutically acceptable salts and solvates thereof, However, the following conditions apply: a)R 1a represents Het, the 5- or 6-membered monocyclic aromatic ring is 3~6 Cycloalkyl and C 1~4 is substituted with three substituents selected from the group consisting of alkyl; b)R 1a -C(=O)-NR xa R xb R xa But one -OH, one -OC 1~4 Alkyl or NR 11c R 11d C replaced with 1~4 C substituted with alkyl and 1, 2 or 3 halo atoms 1~4 is selected from the group consisting of alkyl, c)R 1c represents the halo, d) R 4 is other than isopropyl, e)R 3 But -C 1~6 Alkyl-NR 8a R8b R 8a But -C≡C, -CH=CH, -SC 1~4 Alkyl, -S(=O)2-NR 11a R 11b , 1 NR 11a R 11b -OC is substituted with 1~4 -OC substituted with alkyl and 1, 2 or 3 halo atoms 1~4 C substituted with 1, 2 or 3 substituents each independently selected from the group consisting of alkyl 1~6 is alkyl.

[0013] The present invention also relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I), a pharma- ceutically acceptable salt or solvate thereof, and a pharma- ceutically acceptable carrier or excipient.

[0014] The present invention further relates to the compounds of formula (I), pharma- ceutically acceptable salts or solvates thereof for use as a medicament, and to the compounds of formula (I), pharma- ceutically acceptable salts or solvates thereof for use in the treatment or prevention of cancer, including but not limited to leukemia, myelodysplastic syndromes (MDS), and myeloproliferative neoplasms (MPN), and glycosis.

[0015] In a particular embodiment, the present invention relates to a compound of formula (I), a pharma- ceutically acceptable salt or solvate thereof, for use in the treatment of cancer or in the prevention of cancer.

[0016] In a specific embodiment, said cancer is selected from leukemia, lymphoma, myeloma, or solid tumor cancer (such as, for example, prostate cancer, lung cancer, breast cancer, pancreatic cancer, colon cancer, liver cancer, melanoma, and glioblastoma). In some embodiments, the leukemia is selected from the group consisting of acute leukemia, chronic leukemia, myelocytic leukemia, myelogenous leukemia, lymphoblastic leukemia, lymphocytic leukemia, acute myelogenous leukemias (AML), chronic myelogenous leukemias (CML), acute lymphoblastic leukemias (ALL), chronic lymphocytic leukemias (CLL), T cell prolymphocytic leukemias (T-PLL), large granular lymphocytic leukemia, hairy cell leukemia, and / or idiopathic leukemia. These include MLL-rearranged leukemia, MLL-PTD leukemia, MLL-amplified leukemia, MLL-positive leukemia, and leukemias that exhibit a HOX / MEIS1 gene expression signature.

[0017] In particular, the compounds according to the invention and pharmaceutical compositions thereof may be useful in the treatment or prevention of leukemia, in particular nucleophosmin (NPM1) mutant leukemia, such as NPM1c.

[0018] In certain embodiments, the compounds of formula (I) and their pharma- ceutically acceptable salts and solvates may have improved metabolic stability properties.

[0019] In certain embodiments, the compounds of formula (I) and their pharma- ceutically acceptable salts and solvates may have an extended in vivo half-life (T1 / 2).

[0020] In certain embodiments, the compounds of formula (I) and their pharma- ceutically acceptable salts and solvates may have improved oral bioavailability.

[0021] In certain embodiments, the compounds of formula (I) and their pharma- ceutically acceptable salts and solvates may reduce tumor growth, for example, tumors harboring MLL (KMT2A) gene rearrangements / alterations and / or NPM1 mutations.

[0022] In certain embodiments, compounds of formula (I) and their pharma- ceutically acceptable salts and solvates may have improved in vivo PD properties over an extended period of time, e.g., inhibition of target gene expression, such as MEIS1, and upregulation of differentiation markers for at least 16 hours.

[0023] In certain embodiments, the compounds of formula (I) and their pharma- ceutically acceptable salts and solvates may have an improved safety profile (e.g., reduced hERG inhibition, improved cardiovascular safety).

[0024] In certain embodiments, the compounds of formula (I) and their pharma- ceutically acceptable salts and solvates may be suitable for QD administration (once per day).

[0025] The present invention also relates to the use of a compound of formula (I), a pharma- ceutically acceptable salt or solvate thereof, in combination with an additional pharmaceutical agent for use in the treatment or prevention of cancer, including, but not limited to, leukemia, myelodysplastic syndromes (MDS), and myeloproliferative neoplasms (MPN), and glycosis.

[0026] Furthermore, the present invention relates to a process for preparing a pharmaceutical composition according to the present invention, characterized in that a pharma- ceutically acceptable carrier is intimately mixed with a therapeutically effective amount of a compound of formula (I), a pharma- ceutically acceptable salt or solvate thereof.

[0027] The present invention relates to a product comprising a compound of formula (I), a pharma- ceutically acceptable salt, or solvate thereof, and an additional medicinal agent as a combined preparation for simultaneous, separate or sequential use, for use in the treatment or prevention of cancer and glycological diseases, including leukemia, myelodysplastic syndromes (MDS), and myeloproliferative neoplasms (MPN).

[0028] Furthermore, the present invention relates to a method for treating or preventing a cell proliferative disorder in a warm-blooded animal, the method comprising administering to said warm-blooded animal an effective amount of a compound of formula (I), a pharma- ceutically acceptable salt or solvate thereof, or a pharmaceutical composition or combination as defined herein. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0029] As used herein, the term "halo" or "halogen" refers to fluoro, chloro, bromo, and iodo.

[0030] As used herein, the prefix "C x~y " (where x and y are integers) refers to the number of carbon atoms in a given group. Thus, C 1~6 The alkyl group contains 1 to 6 carbon atoms, and so forth.

[0031] As used herein as a group or part of a group, "C 1~4 The term "alkyl" refers to a straight or branched chain saturated hydrocarbon radical having from 1 to 4 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, t-butyl, and the like.

[0032] Similarly, when used herein as a group or part of a group, "C 1~6 The term "alkyl" refers to a straight or branched chain saturated hydrocarbon radical having from 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, t-butyl, n-pentyl, n-hexyl, and the like.

[0033] As used herein as a group or part of a group, "C 3~6 The term "cycloalkyl" defines a saturated cyclic hydrocarbon radical having three to six carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0034] It will be apparent to those skilled in the art that S(=O)2 or SO2 represents a sulfonyl moiety.

[0035] It will be apparent to one skilled in the art that CO or C(=O) represent a carbonyl moiety.

[0036] It will be apparent to one of skill in the art that a group such as -CRR represents:

[0037] [ka] Examples of such groups are -CR 5a R 5b -It is.

[0038] Groups such as -NR

[0039] [ka] It will be clear to one skilled in the art that the aryl group represents -NR 5c -It is.

[0040] Non-limiting examples of "monocyclic 5- or 6-membered aromatic ring containing 1, 2, or 3 nitrogen atoms and, optionally, a carbonyl moiety" include, but are not limited to, pyrazolyl, imidazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, or 1,2-dihydro-2-oxo-4-pyridinyl.

[0041] One of ordinary skill in the art will appreciate that 5- or 6-membered monocyclic aromatic rings containing 1, 2, or 3 nitrogen atoms and a carbonyl moiety include, but are not limited to, the following:

[0042] [ka]

[0043] When any variable occurs more than one time in any constituent, each definition is independent.

[0044] When any variable occurs more than one time in any formula (eg, formula (I)), each definition is independent.

[0045] In general, whenever the term "substituted" is used in the present invention, unless otherwise indicated or apparent from the context, it is meant to indicate that one or more hydrogens, particularly 1 to 4 hydrogens, more particularly 1 to 3 hydrogens, preferably 1 or 2 hydrogens, more preferably 1 hydrogen on the atom or radical shown in the expression are replaced with a selection from the group shown, provided that the normal valences are not exceeded, and that the replacement results in a chemically stable compound, i.e., a compound that is sufficiently robust to survive isolation from the reaction mixture to a useful degree of purity (post-reaction isolation, for example purification by silica gel chromatography). In certain embodiments, when the number of substituents is not explicitly specified, the number of substituents is 1.

[0046] Combinations of substituents and / or variables are permissible only if such combinations result in chemically stable compounds. "Stable compound," in this context, is meant to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture (post-reaction isolation, e.g., purification by silica gel chromatography).

[0047] Those of skill in the art will understand that the term "optionally substituted" means that the atom or radical designated in the expression using "optionally substituted" may be substituted or unsubstituted (which means substituted or unsubstituted, respectively).

[0048] When two or more substituents are present on a moiety, they may replace hydrogen atoms on the same atom or they may replace hydrogen atoms on different atoms in the moiety, unless otherwise stated or apparent from the context.

[0049] Within the context of the present invention, "saturated" means "fully saturated" unless otherwise stated.

[0050] Unless otherwise stated or apparent from the context, the aromatic ring group may be attached to the remainder of the molecule of formula (I) by any available ring carbon atom (C-bonded) or nitrogen atom (N-bonded).

[0051] Unless otherwise specified or apparent from the context, aromatic ring groups may be optionally substituted on carbon and / or nitrogen atoms where possible according to the embodiment.

[0052] As used herein, the term "subject" refers to an animal, preferably a mammal (e.g., a cat, dog, primate, or human), more preferably a human, who is or has been the object of treatment, observation, or experiment.

[0053] The term "therapeutically effective amount," as used herein, means that amount of an active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue system, animal, or human that is sought by a researcher, veterinarian, physician, or other clinician, including the alleviation or reversal of symptoms of the disease or disorder being treated.

[0054] The term "composition" is intended to encompass a product comprising specified ingredients in specified amounts, and any product that results directly or indirectly from a combination of the specified ingredients in the specified amounts.

[0055] As used herein, the term "treatment" is intended to refer to any process that can slow, hinder, inhibit, or halt the progression of a disease, although it does not necessarily indicate the complete disappearance of all symptoms.

[0056] As used herein, the term "compounds of the invention" or "compounds according to the invention" is meant to include compounds of formula (I) and their pharma- ceutically acceptable salts and solvates.

[0057] As used herein, any chemical formula with bonds shown only as solid lines and not as solid wedge bonds or hashed wedge bonds, or otherwise shown as having a particular configuration (e.g., R, S) around one or more atoms contemplates each possible stereoisomer, or a mixture of two or more stereoisomers.

[0058] In the above and below, the term "compounds of formula (I)" is meant to include their tautomers and their stereoisomeric forms.

[0059] Above and below, the terms "stereoisomer", "stereoisomeric form" or "stereochemically isomeric form" are used interchangeably.

[0060] The present invention includes all stereoisomers of the compounds of the present invention, either as a pure stereoisomer or as a mixture of two or more stereoisomers.

[0061] Enantiomers are stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a racemate or racemic mixture.

[0062] Atropisomers (or atropoisomers) are stereoisomers with specific spatial configurations resulting from restricted rotation about a single bond due to significant steric hindrance. All atropisomers of the compounds of formula (I) are intended to be included within the scope of the present invention.

[0063] Diastereomers (or diastereoisomers) are stereoisomers that are not enantiomers, i.e., they are not related as mirror images. When a compound contains double bonds, the substituents may be in the E or Z configuration.

[0064] Substituents on a divalent cyclic saturated or partially saturated radical can have either the cis or trans configuration; for example, if the compound contains a disubstituted cycloalkyl group, the substituents can be in either the cis or trans configuration.

[0065] Thus, the present invention includes enantiomers, atropisomers, diastereomers, racemates, E isomers, Z isomers, cis isomers, trans isomers, and mixtures thereof, whenever chemically possible.

[0066] The meanings of all terms, i.e. enantiomers, atropisomers, diastereomers, racemates, E isomers, Z isomers, cis isomers, trans isomers and mixtures thereof, are known to the person skilled in the art.

[0067] Absolute configuration is specified according to the Cahn-Ingold-Prelog system. The configuration at the asymmetric atom is specified by either R or S. Resolved stereoisomers for which the absolute configuration is not known are They can be designated (+) or (-) depending on the direction they rotate plane-polarized light. For example, resolved enantiomers whose absolute configuration is not known can be designated (+) or (-) depending on the direction they rotate plane-polarized light.

[0068] When a particular stereoisomer is specified, this means that said stereoisomer is substantially free of other stereoisomers, i.e. associated with less than 50%, preferably less than 20%, more preferably less than 10%, even more preferably less than 5%, particularly less than 2%, most preferably less than 1% of other stereoisomers. Thus, when a compound of formula (I) is specified, for example, as (R), this means that the compound is substantially free of (S) isomers, when a compound of formula (I) is specified, for example, as E, this means that the compound is substantially free of Z isomers, and when a compound of formula (I) is specified, for example, as cis, this means that the compound is substantially free of trans isomers.

[0069] Some of the compounds according to formula (I) may also exist in their tautomeric forms. Although not explicitly shown in formula (I) above, such forms, to the extent that they may exist, are intended to be included within the scope of the present invention. Thus, a single compound may exist in both stereoisomeric and tautomeric forms.

[0070] Pharmaceutically acceptable salts include acid addition salts and base addition salts.Such salts can be formed by conventional means, for example, by reacting free acid form or free base form with one or more equivalents of suitable base or acid, optionally in a solvent or in a medium in which the salt is insoluble, and then removing said solvent or said medium using standard techniques (for example, in vacuum, by lyophilization, or by filtration).Salts can also be prepared by exchanging the counterion of the compound of the present disclosure in the form of a salt with another counterion, for example, by using a suitable ion exchange resin.

[0071] The pharma- ceutically acceptable salts referred to above and hereinafter are meant to include the therapeutically active non-toxic acid and base salt forms which the compounds of formula (I) and their solvates are able to form.

[0072] Suitable acids include, for example, inorganic acids such as hydrohalic acids, e.g., hydrochloric or hydrobromic acids, sulfuric, nitric, phosphoric acids, and the like; or organic acids such as, for example, acetic acid, propanoic acid, hydroxyacetic acid, lactic acid, pyruvic acid, oxalic acid (i.e., ethanedioic acid), malonic acid, succinic acid (i.e., butanedioic acid), maleic acid, fumaric acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclamic acid, salicylic acid, p-aminosalicylic acid, pamoic acid, and the like. Conversely, the above salt forms can be converted to the free base form by treatment with an appropriate base.

[0073] The compounds of formula (I) or solvates thereof containing acidic protons may be converted into their non-toxic metal or amine salt forms by treatment with appropriate organic and inorganic bases.

[0074] Suitable base salt forms include, for example, ammonium salts, alkali and alkaline earth metal salts such as lithium, sodium, potassium, cesium, magnesium, calcium salts, and the like, salts with organic bases such as primary, secondary and tertiary aliphatic amines and aromatic amines such as methylamine, ethylamine, propylamine, isopropylamine, the four butylamine isomers, dimethylamine, diethylamine, diethanolamine, dipropylamine, diisopropylamine, di-n-butylamine, pyrrolidine, piperidine, morpholine, trimethylamine, triethylamine, tripropylamine, quinuclidine, pyridine, quinoline, and isoquinoline; benzathine, N-methyl-glucamine, hydrabamine salts, and salts with amino acids such as arginine, lysine, and the like. Conversely, the base forms can be converted to the free base forms by treatment with acid.

[0075] The term "prodrug" includes any compound which, following oral or parenteral administration, in particular oral administration, is metabolized (to a (more) active) form (in vivo) in an experimentally detectable amount and within a given time, for example within a dosing interval of 0.5 to 24 hours, or for example within a dosing interval of 6 to 24 hours, i.e. 1 to 4 times per day. For the avoidance of doubt, the term "parenteral" administration includes all forms of administration other than oral administration, in particular intravenous (IV), intramuscular (IM) and subcutaneous (SC) injection.

[0076] Prodrugs can be prepared by modifying functional groups present on a compound such that the modification is cleaved in vivo when such prodrug is administered to a mammalian subject. The modification is typically accomplished by synthesizing the parent compound with a prodrug substituent. In some embodiments, prodrugs include compounds in which a hydroxyl, amino, sulfhydryl, carboxy, or carbonyl group is bonded to any group that can be cleaved in vivo to regenerate a free hydroxyl, amino, sulfhydryl, carboxy, or carbonyl group, respectively.

[0077] Examples of prodrugs include, but are not limited to, esters and carbamates of hydroxyl functional groups, ester groups of carboxyl functional groups, N-acyl derivatives and N-Mannich bases. General information on prodrugs can be found, for example, in Bundegaard, H. "Design of Prodrugs" pl-92, Elsevier, New York-Oxford (1985).

[0078] The term solvates comprises the solvent addition forms, which the compounds of formula (I) are able to form, as well as the salts thereof. Examples of such solvent addition forms are, for example, hydrates, alcoholates, etc.

[0079] The compounds of the present invention prepared by the process described below may be synthesized in the form of a mixture of enantiomers, in particular a racemic mixture of enantiomers, which can be separated from each other according to resolution procedures known in the art. Methods for separating the enantiomeric forms of the compound of formula (I) and its pharma-ceutically acceptable salts and solvates include liquid chromatography using chiral stationary phases. The pure stereochemical isomers may also be derived from the corresponding pure stereochemical isomers of the appropriate starting materials, provided that the reaction occurs stereospecifically. Preferably, when a specific stereoisomer is desired, the compound will be synthesized by stereospecific preparation methods. These methods will advantageously employ enantiomerically pure starting materials.

[0080] As used herein, the term "enantiomerically pure" means that the product contains at least 80% by weight of one enantiomer and no more than 20% by weight of the other enantiomer. Preferably, the product contains at least 90% by weight of one enantiomer and no more than 10% by weight of the other enantiomer. In the most preferred embodiment, the term "enantiomerically pure" means that the composition contains at least 99% by weight of one enantiomer and no more than 1% of the other enantiomer.

[0081] The present invention also includes isotopically labeled compounds of the present invention which are identical to those enumerated herein, but by virtue of the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number normally found in nature (or the most abundant atom found in nature).

[0082] All isotopes and isotopic mixtures of any particular atom or element identified herein, whether naturally occurring or synthetically produced, either at natural abundance or in isotopically enriched form, are contemplated within the scope of the compounds of the invention. Exemplary isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine; 2 H, 3 H, 11 C. 13 C. 14 C. 13 N, 15 O. 17 O. 18 O. 32 P, 33 P, 35 S, 18 F, 36 Cl, 122 I, 123 I, 125 I, 131 I, 75 Br, 76 Br, 77 Br, and 82Br. Preferably, the isotope is 2 H, 3 H, 11 C. 13 C, and 18 F. Preferably, the isotope is selected from the group 2 H, 3 H, 11 C, and 18 F. More preferably, the isotope is selected from the group 2 H, 3 H or 13 C. More preferably, the isotope is 2 H or 13 C. More preferably, the isotope is 2 H. In particular, deuterium compounds and 13 C-enriched compounds are intended to be included within the scope of the present invention. In particular, deuterium compounds are intended to be included within the scope of the present invention.

[0083] Certain isotopically labeled compounds of the invention (e.g., 3 H and 14 C) can be useful, for example, in substrate tissue distribution assays. 3 H) and carbon-l4( 14 C) isotopes are useful for their ease of preparation and detectability. Additionally, heavier isotopes, such as deuterium (i.e., 2 H), may result in greater metabolic stability (e.g., increased half-life in vivo or reduced dosage requirements), which may result in certain therapeutic advantages and therefore may be preferred in some circumstances. For example, 15 O. 13 N, 11 C, and 18Positron-emitting isotopes such as F are useful in positron emission tomography (PET) studies. PET imaging in cancer finds utility in helping to localize and identify tumors, stage disease, and determine suitable treatments. Human cancer cells overexpress many receptors or proteins that are potential disease-specific molecular targets. Radiolabeled tracers that bind with high affinity and specificity to such receptors or proteins on tumor cells have great potential for diagnostic imaging and targeted radionuclide therapy (Charron, Carlie L. et al. Tetrahedron Lett. 2016, 57(37), 4119-4127). Furthermore, target-specific PET radiotracers can be used as biomarkers to investigate and evaluate pathology, for example, by measuring target expression and treatment response (Austin R. et al. Cancer Letters (2016), doi:10.1016 / j.canlet.2016.05.008).

[0084] The present invention relates in particular to compounds of formula (I) as defined herein, R 1a -C(=O)-NR xa R xb , Het, or

[0085] [ka] represents Het represents a 5- or 6-membered monocyclic aromatic ring containing 1, 2 or 3 nitrogen atoms and, optionally, a carbonyl moiety; The 5- or 6-membered monocyclic aromatic ring is 3~6 Cycloalkyl and C 1~4 optionally substituted with 1, 2 or 3 substituents selected from the group consisting of alkyl; R xa and R xb are each independently hydrogen; C 1~4 Alkyl;C 3~6Cycloalkyl; C substituted with 1, 2 or 3 halo atoms 1~4 Alkyl; and one -OH, -OC 1~4 Alkyl or NR 11c R 11d C replaced with 1~4 alkyl; R 1b represents F or Cl, R 1c represents H or halo; Y 1 represents -O-; R 2 represents hydrogen, U represents N; n1, n2, n3 and n4 are each independently selected from 1 and 2; X 1 represents CH, and X 2 represents N, R 4 But, C 1~5 Alkyl,

[0086] [ka] represents R 3 But, C 1~6 Alkyl-NR 8a R 8b represents R 3 C in the definition of 1~6 The alkyl portion is selected from the group consisting of cyano, halo, -OH, and -OC 1~4 and optionally substituted with 1, 2, or 3 substituents each independently selected from the group consisting of alkyl, R 8a and R 8b are each independently hydrogen; C 1~6 Alkyl; -C(=O)-C 1~4 Alkyl; -C(=O)-OC 1~4 Alkyl; -C(=O)-NR 12a R 12b ; and -OH, cyano, halo, -C≡C, -CH=CH, -SC 1~4Alkyl, -S(=O)2-C 1~4 Alkyl, -S(=O)2-NR 11a R 11b , -OC 1~4 Alkyl, -C(=O)-NR 10a R 10b , -NR 10c -C(=O)-C 1~4 Alkyl, 1 NR 11a R 11b -OC is substituted with 1~4 -OC substituted with alkyl and 1, 2 or 3 halo atoms 1~4 C substituted with 1, 2 or 3 substituents each independently selected from the group consisting of alkyl 1~6 alkyl; R 10a , R 10b , R 10c , R 11a , R 11b , R 12a , and R 12b each independently represents hydrogen and C 1~6 is selected from the group consisting of alkyl, R 11c and R 11d each independently represents hydrogen, C 1~6 Alkyl and -C(=O)-C 1~4 and tautomeric and stereoisomeric forms thereof; and pharma- ceutically acceptable salts and solvates thereof, However, the following conditions apply: a)R 1a represents Het, the 5- or 6-membered monocyclic aromatic ring is 3~6 Cycloalkyl and C 1~4 is substituted with three substituents selected from the group consisting of alkyl; b)R 1a -C(=O)-NR xa R xb R xa But one -OH, one -OC 1~4 Alkyl or NR 11c R 11d C replaced with1~4 Alkyl; and C substituted with 1, 2 or 3 halo atoms 1~4 alkyl; c)R 1c represents the halo, d) R 4 is other than isopropyl, e)R 3 But -C 1~6 Alkyl-NR 8a R 8b R 8a But -C≡C, -CH=CH, -SC 1~4 Alkyl, -S(=O)2-NR 11a R 11b , 1 NR 11a R 11b -OC is substituted with 1~4 -OC substituted with alkyl and 1, 2 or 3 halo atoms 1~4 C substituted with 1, 2 or 3 substituents each independently selected from the group consisting of alkyl 1~6 is alkyl.

[0087] The present invention relates in particular to compounds of formula (I) as defined herein, R 1a -C(=O)-NR xa R xb represents R xa and R xb However, each independently, C 1~4 alkyl; and one -OH or -NR 11c R 11d C replaced with 1~4 alkyl; R 1b represents F, R 1c represents H or halo; Y 1 represents -O-; R 2 represents hydrogen, n1, n2, n3 and n4 are each independently selected from 1 and 2; X 1 represents CH, and X 2 represents N, R 4 But, C 1~5 represents an alkyl group, R 3 But -C 1~6 Alkyl-NR 8a R 8b represents R 3 C in the definition of 1~6 the alkyl portion is optionally substituted with 1, 2 or 3 -OH substituents; R 8a and R 8b are each independently hydrogen; C 1~6 Alkyl; and -C≡C, -CH=CH, -SC 1~4 Alkyl, -S(=O)2-NR 11a R 11b , -OC 1~4 Alkyl, 1 NR 11a R 11b -OC is substituted with 1~4 -OC substituted with alkyl and 1, 2 or 3 halo atoms 1~4 C substituted with 1, 2 or 3 substituents each independently selected from the group consisting of alkyl 1~6 alkyl; R 11a and R 11b represents hydrogen, R 11c and R 11d are each independently hydrogen and -C(=O)-C 1~4 and tautomeric and stereoisomeric forms thereof; and pharma- ceutically acceptable salts and solvates thereof, However, the following conditions apply: a)R 1a -C(=O)-NR xa R xb R xa has one -OH or NR 11c R 11d C replaced with 1~4alkyl; b)R 1c represents the halo, c)R 4 , tert-butyl; d) R 3 But -C 1~6 Alkyl-NR 8a R 8b R 8a But -C≡C, -CH=CH, -SC 1~4 Alkyl, -S(=O)2-NR 11a R 11b , 1 NR 11a R 11b -OC is substituted with 1~4 -OC substituted with alkyl and 1, 2 or 3 halo atoms 1~4 C substituted with 1, 2 or 3 substituents each independently selected from the group consisting of alkyl 1~6 is alkyl.

[0088] In certain embodiments, the present invention relates to compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R 1b represents F.

[0089] In certain embodiments, the present invention relates to compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R 2 represents hydrogen.

[0090] In certain embodiments, the present invention relates to those compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein n1 is 1, n2 is 2, n3 is 1, and n4 is 1.

[0091] In certain embodiments, the present invention relates to compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein Y 1 represents -O-.

[0092] In certain embodiments, the present invention relates to those compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein: Y 1 represents -O-, U represents N.

[0093] In certain embodiments, the present invention relates to those compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein U represents N.

[0094] In certain embodiments, the present invention relates to those compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein: Y 1 represents -O-, U represents N; R 1b represents F, R 2 represents hydrogen.

[0095] In certain embodiments, the present invention relates to those compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein: R 1a is -C(=O)-NR xa R xb , Y 1 represents -O-, U represents N; R 1b represents F, R1c represents H, R 2 represents hydrogen, R 4 is C 1~5 represents an alkyl group, R 3 is C 1~6 Alkyl-NR 8a R 8b Represents.

[0096] In certain embodiments, the present invention relates to those compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein: R 1a is -C(=O)-NR xa R xb , Y 1 represents -O-, U represents N; n1 is 1, n2 is 2, n3 is 1, and n4 is 1.

[0097] R 1b represents F, R 1c represents H, R 2 represents hydrogen, R 4 is C 1~5 represents an alkyl group, R 3 is C 1~6 Alkyl-NR 8a R 8b Represents.

[0098] In certain embodiments, the present invention relates to compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R 1a is -C(=O)-NR xa R xb Represents.

[0099] In certain embodiments, the present invention relates to compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R 3 -C 1~6 Alkyl-NR 8a R 8b Represents.

[0100] In certain embodiments, the present invention relates to compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R 3 is -CH2-CH2-CH2-NR 8a R 8b Represents.

[0101] In certain embodiments, the present invention relates to compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R 3 C in the definition of 1~6 Alkyl-C 1~6 Alkyl-NR 8a R 8b is limited to -CH2-CH2-CH2-.

[0102] In certain embodiments, the present invention relates to those compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, provided that the compound satisfies the following provisos: a)R 1a -C(=O)-NR xa R xb R xa But one -OH, one -OC 1~4 Alkyl or NR 11c R 11d C replaced with 1~4 C substituted with alkyl and 1, 2 or 3 halo atoms 1~4 is selected from the group consisting of alkyl, b)R 1c represents the halo, R4 is other than isopropyl, d) R 3 But -C 1~6 Alkyl-NR 8a R 8b R 8a But -C≡C, -CH=CH, -SC 1~4 Alkyl, -S(=O)2-NR 11a R 11b , 1 NR 11a R 11b -OC is substituted with 1~4 -OC substituted with alkyl and 1, 2 or 3 halo atoms 1~4 C substituted with 1, 2 or 3 substituents each independently selected from the group consisting of alkyl 1~6 is alkyl.

[0103] In certain embodiments, the present invention relates to compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R 1a represents Het, the 5- or 6-membered monocyclic aromatic ring is C 3~6 Cycloalkyl and C 1~4 and is substituted with three substituents selected from the group consisting of alkyl.

[0104] In certain embodiments, the present invention relates to compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R 1a is -C(=O)-NR xa R xb R xa is one -OH, -OC 1~4 Alkyl or NR 11c R 11d C replaced with 1~4 C substituted with alkyl and 1, 2 or 3 halo atoms 1~4 alkyl.

[0105] In certain embodiments, the present invention relates to compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R 1c represents a halo.

[0106] In certain embodiments, the present invention relates to compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R 1c represents Br.

[0107] In certain embodiments, the present invention relates to compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R 4 represents another isopropyl.

[0108] In certain embodiments, the present invention relates to compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R 4 represents tert-butyl.

[0109] In certain embodiments, the present invention relates to compounds of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R 3 -C 1~6 Alkyl-NR 8a R 8b R 8a -C≡C, -CH=CH, -SC 1~4 Alkyl, -S(=O)2-NR 11a R 11b , 1 NR 11a R 11b OC replaced by 1~4 -OC substituted with alkyl and 1, 2 or 3 halo atoms 1~4C substituted with 1, 2 or 3 substituents each independently selected from the group consisting of alkyl 1~6 It is an alkyl.

[0110] In certain embodiments, the present invention relates to a subgroup of Formula (I) defined in the General Reaction Scheme.

[0111] In certain embodiments, the compound of formula (I) is any of the exemplified compounds: its tautomers and its stereoisomeric forms, and the free base, any pharma- ceutically acceptable salt, and solvate thereof.

[0112] All possible combinations of the above embodiments are considered to be within the scope of the present invention.

[0113] Processes for the preparation of compounds of formula (I) In this section, and in all other sections, unless the context indicates otherwise, references to formula (I) also include all other subgroups and embodiments thereof defined herein.

[0114] The general preparation of some representative examples of compounds of formula (I) is described below, and in certain examples, they are generally prepared from starting materials that are either commercially available or prepared by standard synthetic processes commonly used by those skilled in the art of organic chemistry. The following schemes are merely illustrative of examples of the present invention and are not intended to limit the present invention in any way.

[0115] Alternatively, the compounds of the present invention may also be prepared by analogous reaction protocols as depicted in the following general schemes in combination with standard synthetic processes commonly used by those skilled in the art.

[0116] Those skilled in the art will appreciate that in the reactions depicted in the schemes, although this is not always explicitly shown, it may be necessary to protect reactive functional groups (e.g., hydroxy, amino, or carboxy groups) if these are desired in the final product to avoid their undesired participation in the reaction. In general, conventional protecting groups (PG) can be used in accordance with standard practice. The protecting groups can be removed at a subsequent convenient stage using methods well known in the art.

[0117] Those skilled in the art will appreciate that in the reactions depicted in the schemes, it may be advisable or necessary to carry out the reactions under an inert atmosphere, such as, for example, under an N2-gas atmosphere.

[0118] It will be apparent to one skilled in the art that it may be necessary to cool the reaction mixture prior to working up the reaction (e.g., referring to a series of operations required to isolate and purify the products of a chemical reaction, such as quenching, column chromatography, extraction, etc.).

[0119] Those skilled in the art will appreciate that heating the reaction mixture under stirring may enhance the reaction outcome. In some reactions, microwave heating may be used instead of conventional heating to reduce the overall reaction time.

[0120] Those skilled in the art will appreciate that the alternative series of chemical reactions shown in the schemes below may also lead to the desired compounds of formula (I).

[0121] Those skilled in the art will appreciate that the intermediates and final compounds shown in the following schemes can be further functionalized according to methods well known to those skilled in the art. The intermediates and compounds described herein can be isolated in free form or as a salt or solvate thereof. The intermediates and compounds described herein can be synthesized in the form of a mixture of tautomeric and stereoisomeric forms, which can be separated from one another according to resolution procedures known in the art.

[0122] General synthesis scheme All abbreviations used in the general schemes are as defined in the tables in the Examples part. Variables are as defined in the ranges or as specifically defined in the general schemes.

[0123] Part A) Schemes 1a, 1b, 1c, 2a, 2b and 3

[0124] [ka]

[0125] In Schemes 1a, 1b, and 1c, the following reaction conditions apply: Step 1: Using a suitable organometallic reagent, such as isopropylmagnesium bromide, in the presence of a suitable base, such as TMEDA, at a suitable temperature, such as -70°C, in a suitable solvent, such as THF, Step 2: In the presence of a suitable oxidizing reagent, such as DMP, in a suitable solvent, such as DCM, at a suitable temperature, such as 0° C. to room temperature, Step 3: In the presence of a suitable organometallic reagent, such as isopropylmagnesium bromide, in a suitable solvent, such as THF, at a suitable temperature, such as from −20° C. to room temperature, Step 4: At a suitable temperature, e.g. 80° C., in the presence of a suitable base, e.g. NaOH, in a suitable solvent, e.g. THF and HO Step 5: At a suitable temperature, such as room temperature, in the presence of a suitable amide condensation reagent, such as EDCI and HOBt, in the presence of a suitable base, such as NMM, in a suitable solvent, such as DCM. Step 6: In the presence of a suitable organometallic reagent, such as isopropyllithium, in a suitable solvent, such as THF, at a suitable temperature, such as -70°C, Step 7: At a suitable temperature, such as 90° C., in the presence of a suitable organometallic catalyst, such as Pd(dppf)Cl, in the presence of a suitable base, such as NaCO, in a suitable solvent, such as 1,4-dioxane and H0 Step 8: In the presence of a suitable Lewis acid, such as BBr3, in a suitable solvent, such as DCM, at a suitable temperature, such as 0° C. to room temperature, Step 9: In the presence of a suitable base, such as TEA, DBU or K2CO3, in a suitable solvent, such as DCM, THF or DMF, at a suitable temperature, such as from -78°C to 40°C, in particular from 0°C to room temperature,

[0126] [ka]

[0127] [ka]

[0128] In Schemes 2a and 2b, the following reaction conditions apply: Step 9: See Scheme 1, Step 9.

[0129] Step 10: At a suitable temperature, e.g. room temperature, in the presence of a suitable catalyst, e.g. Pd / C, in the presence of a suitable reducing agent, e.g. H2, optionally in the presence of a suitable base, e.g. TEA, in a suitable solvent, e.g. THF, Alternatively, it can be carried out at a suitable temperature, for example room temperature, in the presence of a suitable solvent, for example Pd(dppf)Cl2·DCM complex, a suitable reducing agent, for example NaBH4, a suitable base, for example TMEDA, in a suitable solvent, for example THF.

[0130] Step 11: for N-deprotection, at a suitable temperature, for example room temperature, in the presence of a suitable acid, for example TFA, in a suitable solvent, for example DCM; for O-deprotection, at a suitable temperature, for example room temperature, in the presence of a suitable acid, for example 4-methylbenzenesulfonic acid, in a suitable solvent, for example MeOH; Step 12: At a suitable temperature, e.g. 80° C., optionally in the presence of a suitable Lewis acid, e.g. ZnCl2, in the presence of a suitable reducing agent, e.g. NaBH3CN, in a suitable solvent, e.g. MeOH, Step 13: At a suitable temperature, such as room temperature, in the presence of a suitable organometallic catalyst, such as Ag(Phen)2OTf, in the presence of a suitable brominating reagent, such as 1,3-dibromo-1,3,5-triazinane-2,4,6-trione, in a suitable solvent, such as DCE, Step 14: This can be carried out at a suitable temperature, such as room temperature, in the presence of a suitable chlorinating reagent, such as oxalyl chloride, in the presence of DMF, in a suitable solvent, such as DCM.

[0131] [ka]

[0132] In Scheme 3, the following reaction conditions apply: Steps 11-12: See steps 11-12 in Scheme 2.

[0133] Step 15: At a suitable temperature, for example 80° C., in the presence of a suitable base, for example Cs2CO3, in a suitable solvent, for example DMF Step 16: can be carried out at a suitable temperature, for example 40° C., in the presence of a suitable base, for example ammonia, in a suitable solvent, for example 1,4-dioxane.

[0134] Part B) Schemes 4, 5, 6, 7, 8, 9, 10, 11 and 12

[0135] [ka]

[0136] In Scheme 4, the following reaction conditions apply: Step 1: At a suitable temperature, e.g. 90° C., in the presence of a suitable organometallic catalyst, e.g. Pd(dppf)Cl2, in the presence of a suitable base, e.g. Na2CO3, in a suitable solvent, e.g. 1,4-dioxane and HO Step 2: At a suitable temperature, such as room temperature, in the presence of a suitable amide condensation reagent, such as HATU, in the presence of a suitable base, such as DIEA, in a suitable solvent, such as DCM, Step 3: In the presence of a suitable Lewis acid, such as BBr3, in a suitable solvent, such as DCM, at a suitable temperature, such as -78°C to room temperature Step 4: In the presence of a suitable base, such as TEA, DBU or K2CO3, in a suitable solvent, such as DCM, THF or DMF, at a suitable temperature, such as, for example, from -78°C to 40°C, in particular from 0°C to room temperature, Step 5: At a suitable temperature, for example room temperature, in the presence of a suitable base, for example LiOH·HO, in a suitable solvent, for example THF and HO, Step 6: At a suitable temperature, such as room temperature, in the presence of a suitable organometallic catalyst, such as Ag(Phen)2OTf, in the presence of a suitable brominating reagent, such as 1,3-dibromo-1,3,5-triazinane-2,4,6-trione, in a suitable solvent, such as DCE, Step 7: can be carried out at a suitable temperature, such as room temperature, in the presence of a suitable brominating reagent, such as 1,3-dibromo-1,3,5-triazinane-2,4,6-trione, in the presence of 2,2,2-trifluoroethan-1-ol as solvent.

[0137] [ka]

[0138] In Scheme 5, the following reaction conditions apply: Step 8: In the presence of a suitable base, such as TEA, DBU or K2CO3, in a suitable solvent, such as DCM, THF or DMF, at a suitable temperature, such as from -78°C to 40°C, in particular from 0°C to room temperature, Step 9: In the presence of a suitable base, such as TEA, DBU or K2CO3, in a suitable solvent, such as DCM, THF or DMF, at a suitable temperature, such as from -78°C to 40°C, in particular from 0°C to room temperature, Step 10: At a suitable temperature, e.g. room temperature, in the presence of a suitable organometallic catalyst, e.g. Pd / C, and a suitable base, e.g. TEA, in a suitable solvent, e.g. MeOH, under an atmosphere of H2; Step 11: When PG is Boc, it can be carried out at a suitable temperature, such as room temperature, in the presence of a suitable acid, such as TFA, in a suitable solvent, such as DCM.

[0139] [ka]

[0140] In Scheme 6, the following reaction conditions apply: Step 12: Reductive amination conditions, at a suitable temperature, for example room temperature to 80° C., in the presence or absence of a suitable Lewis acid, for example ZnCl2, or an acid, for example AcOH, or in the presence of a suitable reducing agent, for example NaBH3CN, in a suitable solvent, for example MeOH; Step 13: At a suitable temperature, e.g. 0° C., in the presence of a suitable electrophile, e.g. MsCl, in the presence of a suitable base, e.g. TEA, in a suitable solvent, e.g. DCM Step 14: In the presence of a suitable oxidizing agent, such as DMP, in a suitable solvent, such as DCM, at a suitable temperature, such as 0° C. to room temperature, Step 15: In the presence of a suitable acid, such as HCl, in a suitable solvent, such as ACN, at a suitable temperature, such as 50° C. Step 16: This can be carried out at a suitable temperature, such as room temperature, in the presence or absence of a suitable base, such as TEA, in a suitable solvent, such as THF.

[0141] [ka]

[0142] In Scheme 7, the following reaction conditions apply: Step 11: When PG is Boc, at a suitable temperature, for example room temperature, in the presence of a suitable acid, for example TFA, in a suitable solvent, for example DCM, Step 12: Reductive amination conditions, at a suitable temperature, for example room temperature to 80° C., in the presence or absence of a suitable Lewis acid, for example ZnCl2, or an acid, for example AcOH, or in the presence of a suitable reducing agent, for example NaBH3CN, in a suitable solvent, for example MeOH; Step 17: In the presence of a suitable base, such as DIEA or Cs2CO3, in a suitable solvent, such as DCM or DMF, at a suitable temperature, such as room temperature to 80°C. Step 18: can be carried out at a suitable temperature, for example 40° C., in the presence of a suitable base, for example ammonia, in a suitable solvent, for example 1,4-dioxane.

[0143] [ka]

[0144] In Scheme 8, the following reaction conditions apply: Step 9: In the presence of a suitable base, such as TEA, DBU or K2CO3, in a suitable solvent, such as DCM, THF or DMF, at a suitable temperature, such as from -78°C to 40°C, in particular from 0°C to room temperature, Step 10: At a suitable temperature, e.g. room temperature, in the presence of a suitable organometallic catalyst, e.g. Pd / C, optionally in the presence of a suitable base, e.g. TEA, in a suitable solvent, e.g. MeOH under an H2 atmosphere; Step 19: At a suitable temperature, such as room temperature, in the presence of a suitable chlorinating reagent, such as oxalyl chloride, in the presence of DMF, in a suitable solvent, such as DCM, Step 20: At a suitable temperature, e.g. 90° C., in the presence of a suitable nucleophilic amine, in a suitable solvent, e.g. EtOH, Step 21: In the presence of a suitable acid, HCl in dioxane, in a suitable solvent, for example MeOH, at a suitable temperature, for example room temperature, Step 22: This can be carried out at a suitable temperature, e.g. 110° C., in the presence of a suitable boron reagent, e.g. trimethylboroxine, in the presence of a suitable organometallic catalyst, e.g. tetrakis(triphenylphosphine)palladium(0), in the presence of a suitable base, e.g. KCO, in a suitable solvent, e.g. 1,4-dioxane.

[0145] [ka]

[0146] In Scheme 9, the following reaction conditions apply: Step 23: In the presence of a suitable base, such as DIEA and n-BuLi, at a suitable temperature, such as -78°C to -25°C, in the presence of a suitable solvent, such as THF, Step 24: at a suitable temperature, for example -65°C to -55°C, in the presence of a suitable reducing agent, for example DIBAL-H, in a suitable solvent, for example toluene, preferably in a suitable flow chemistry system; Step 25: first in the presence of a suitable base, e.g. DMAP, in the presence of a suitable condensing agent, e.g. DCC, in a suitable solvent, e.g. DCM, at a suitable temperature, e.g. -10°C to 10°C, then in the presence of a suitable acid, e.g. AcOH, in the presence of a suitable reducing agent, e.g. NaBH4, in a suitable solvent, e.g. DCM, at a suitable temperature, e.g. -10°C to 0°C, Step 26: Heat to reflux in a suitable solvent, such as toluene. Step 27: In the presence of a suitable reducing agent, such as LiBH4, in a suitable solvent, such as 2-methyltetrahydrofuran, at a suitable temperature, such as -5°C to 5°C, Step 28: In the presence of a suitable reducing agent, such as NaBH(OAc)3, in a suitable solvent, such as DCM, at a suitable temperature, such as 15°C to 25°C. Step 29: In the presence of a suitable acid, such as HCl, in a suitable solvent, such as IPA, at a suitable temperature, such as 15° C. to 25° C. Step 30: In the presence of a suitable reducing agent, such as NaBH(OAc)3, in the presence of a suitable base, such as TEA, at a suitable temperature, such as 5° C. to 30° C., in a suitable solvent, such as toluene, Step 31: In the presence of a suitable base, such as KHPO, in a suitable solvent, such as HO, at a suitable temperature, such as 50° C. to 55° C. Step 32: When PG is Bn, at a suitable temperature, e.g., −5° C. to 45° C., in a suitable pressure range, e.g., 0.27 to 0.40 MPa, under hydrogen atmosphere, in the presence of a suitable catalyst, e.g., palladium hydroxide on carbon, in the presence of a suitable acid, e.g., MSA, in a suitable solvent, e.g., EtOH; Step 33: In the presence of a suitable base, such as TEA, at a suitable temperature, such as −50° C. to −40° C., in a suitable solvent, such as 2-methyltetrahydrofuran Step 34: In the presence of a suitable base, such as TMG, at a suitable temperature, such as 20° C. to 30° C., in a suitable solvent, such as 2-methyltetrahydrofuran Step 35: At a suitable temperature, e.g. 20° C.-30° C., within a suitable pressure range, e.g. 0.20-0.30 MPa, under hydrogen atmosphere, in the presence of a suitable catalyst, e.g. palladium hydroxide on carbon, in a suitable solvent, e.g. MeOH, Alternatively, it can be carried out at a suitable temperature, for example room temperature, in the presence of a suitable catalyst, for example 1,1'-bis(triphenylphosphine)ferrocene-palladium(II) dichloride dichloromethane complex, a suitable reducing agent, for example sodium borohydride, a suitable base, for example N,N,N',N'-tetramethylethylenediamine, in a suitable solvent, for example tetrahydrofuran.

[0147] Scheme 10 In general, Y 1 is limited to -CH2-, R 2 W 1 Compounds of formula (I) limited to, herein referred to as compounds of formula (Ia), can be prepared according to the following reaction scheme 10. In scheme 10, W 1represents chloro, bromo or iodo, and all other variables are defined according to the scope of the present invention.

[0148] [ka]

[0149] In Scheme 10, the following reaction conditions apply: Step 36: can be carried out at a suitable temperature, for example 60° C. to 100° C., in the presence of a suitable catalyst, for example palladium acetate (Pd(OAc)2) or tris(dibenzylideneacetone)dipalladium(0) (Pd2(dba)3) or tetrakis(triphenylphosphine)palladium(0), in a suitable solvent, for example tetrahydrofuran or dioxane.

[0150] Those skilled in the art will appreciate that starting from compound (Ia), chemistry similar to that reported in Scheme 5, step 10 and Scheme 8, steps 20, 21 and 22 can be carried out.

[0151] Scheme 11 In general, Y 1 -CR 5a R 5b Limited to R 2 W 1 Compounds of formula (I) limited to, herein referred to as compounds of formula (Ib), can be prepared according to the following reaction scheme 11. In scheme 11, R 5a and R 5b is other than hydrogen. All other variables are defined in accordance with the scope of the invention.

[0152] [ka]

[0153] In Scheme 11, the following reaction conditions apply: Step 37: At a suitable temperature in the range of 80° C. to 200° C., in the presence of a suitable catalyst such as palladium acetate (Pd(OAc)2), in the presence of a suitable ligand such as triphenylphosphine or tricyclohexylphosphine, in a suitable solvent such as dioxane, preferably under closed conditions, optionally under microwave irradiation.

[0154] Those skilled in the art will appreciate that starting from compound (Ib), chemistry similar to that reported in Scheme 5, step 10 and Scheme 8, steps 20, 21 and 22 can be carried out.

[0155] Scheme 12

[0156] [ka]

[0157] In Scheme 12, the following reaction conditions apply: Step 38: The reaction is carried out at a suitable temperature, such as room temperature to 80° C., in the presence of a suitable base, such as DIEA, Cs 2 CO 3 or DBU, in a suitable solvent, such as DCM, THF or DMF.

[0158] Alternatively, it can be carried out at a suitable temperature, for example, from room temperature to 100° C., in the presence of a suitable catalyst, for example, Pd2dba3, in the presence of a suitable ligand, for example, Xantphos, in the presence of a suitable base, for example, Cs2CO3 or Na2CO3, in a suitable solvent, for example, dioxane or a mixture of dioxane and water.

[0159] Those skilled in the art can start from intermediate A and convert Y 1 It will be appreciated that similar chemistry to that reported can be performed when represents O.

[0160] It will be understood that, where appropriate functional groups are present, the compounds of the various formulae, or any intermediates used in their preparation, may be further derivatized by one or more standard synthetic methods employing condensation, substitution, oxidation, reduction, or cleavage reactions. Particular substitution techniques include conventional alkylation, arylation, heteroarylation, acylation, sulfonylation, halogenation, nitration, formylation, and coupling procedures.

[0161] The compounds of formula (I) may be synthesized in the form of racemic mixtures of enantiomers, which may be separated from each other according to resolution procedures known in the art. Racemic compounds of formula (I) containing a basic nitrogen atom may be converted into the corresponding diastereomeric salt forms by reaction with a suitable chiral acid. The diastereomeric salt forms are then separated, for example, by selective or fractional crystallization, and the enantiomers are liberated therefrom by alkali. An alternative method for separating the enantiomeric forms of the compounds of formula (I) includes liquid chromatography using a chiral stationary phase. The pure stereochemically isomers may also be derived from the corresponding pure stereochemically isomers of the appropriate starting materials, provided that the reaction occurs stereospecifically.

[0162] In the preparation of the compounds of the present invention, protection of remote functional groups (e.g., primary or secondary amines) of intermediates may be necessary. The need for such protection will vary with the nature of the remote functional group and the conditions of the preparation method. Suitable amino-protecting groups (NH-Pg) include acetyl, trifluoroacetyl, t-butoxycarbonyl (Boc), benzyloxycarbonyl (CBz), and 9-fluorenylmethyleneoxycarbonyl (Fmoc). The need for such protection is readily determined by one skilled in the art. For a general description of protecting groups and their use, see TW Greene and PG M Huts, Protective Groups in Organic Synthesis, 4th ed., Wiley, Hoboken, New Jersey, 2007.

[0163] Pharmacology It has been found that the compounds of the present invention are capable of blocking the interaction of menin with MLL proteins and oncogenic MLL fusion proteins themselves, or undergoing metabolism in vivo to a (more) active form (prodrug).Accordingly, the compounds according to the present invention and pharmaceutical compositions comprising such compounds may be useful in the treatment or prevention, particularly in the treatment, of diseases such as cancer, including but not limited to leukemia, myelodysplastic syndromes (MDS), and myeloproliferative neoplasms (MPN).

[0164] In particular, the compounds according to the invention and pharmaceutical compositions thereof may be useful in the treatment or prevention of cancer. According to one embodiment, cancers that may benefit from treatment with the menin / MLL inhibitors of the invention include leukemia, lymphoma, myeloma or solid tumor cancers (such as prostate cancer, lung cancer, breast cancer, pancreatic cancer, colon cancer, liver cancer, melanoma and glioblastoma). In some embodiments, leukemias include acute leukemia, chronic leukemia, myeloid leukemia, myeloid leukemia, lymphoblastic leukemia, lymphocytic leukemia, acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), T-cell prolymphocytic leukemia (T-PLL), large granular lymphocytic leukemia, hairy cell leukemia (HCL), MLL-rearranged leukemia, MLL-PTD leukemia, MLL-amplified leukemia, MLL-positive leukemia, leukemias exhibiting a HOX / MEIS1 gene expression signature, and the like.

[0165] In particular, the compounds according to the invention and pharmaceutical compositions thereof may be useful in the treatment or prevention of myelodysplastic syndromes (MDS) or myeloproliferative neoplasms (MPN).

[0166] In particular, the compounds according to the invention and pharmaceutical compositions thereof may be useful in the treatment or prevention of leukemia, in particular nucleophosmin (NPM1) mutant leukemia, such as NPM1c.

[0167] In particular, the compounds according to the present invention and pharmaceutical compositions thereof are useful in the treatment of AML, particularly nucleophosmin (NPM1)-mutated AML (i.e., NPM1 mutAbstract: The present invention may be useful in the treatment or prevention of NPM1-mutated AML, and more specifically, NPM1-mutated AML.

[0168] In particular, the compounds according to the invention and pharmaceutical compositions thereof may be useful in the treatment or prevention of MLL-rearranged leukemia, particularly MLL-rearranged AML or ALL.

[0169] In particular, the compounds according to the present invention and pharmaceutical compositions thereof may be useful in the treatment or prevention of leukemia with MLL gene alterations, particularly AML or ALL with MLL gene alterations.

[0170] In particular, the compounds according to the invention and pharmaceutical compositions thereof may be suitable for QD dosing (once a day).

[0171] In particular, the compounds according to the present invention and pharmaceutical compositions thereof may be useful for treating or preventing hematological cancers in subjects exhibiting NPM1 gene mutations and / or mixed lineage leukemia gene (MLL, MLL1, KMT2A) alterations, mixed lineage leukemia (MLL), MLL-associated leukemia, MLL-associated leukemia, MLL-positive leukemia, MLL-induced leukemia, rearranged mixed lineage leukemia, MLL, rearrangements / alterations or leukemia associated with MLL gene rearrangements / alterations, acute leukemia, chronic leukemia, myelodysplastic syndromes (MDS), myeloproliferative neoplasms (MPN), insulin resistance, pre-diabetes, diabetes or risk of diabetes, hyperglycemia, chromosomal rearrangements on chromosome 11q23, type 1 diabetes, type 2 diabetes, promoting proliferation of pancreatic cells, where the pancreatic cells are islet cells, beta cells, and where the proliferation of beta cells is evidenced by increased beta cell production or insulin production, and inhibiting menin-MLL interaction. Here, the target gene of the MLL fusion protein is HOX or MEIS1 in humans.

[0172] Thus, the present invention relates to compounds of formula (I), their tautomeric and stereoisomeric forms, and their pharma- ceutically acceptable salts and solvates, for use as medicaments.

[0173] The present invention also relates to the use of a compound of formula (I) according to the invention, a tautomeric or stereoisomeric form thereof or a pharma- ceutically acceptable salt or solvate thereof, or a pharmaceutical composition, for the manufacture of a medicament.

[0174] The present invention also relates to a compound of formula (I) according to the present invention, or a tautomeric or stereoisomeric form thereof, or a pharma- ceutically acceptable salt or solvate thereof, or a pharmaceutical composition, for use in the treatment, prevention, amelioration, control or reduction of the risk of a disorder associated with the interaction of menin with MLL proteins and oncogenic MLL fusion proteins in a mammal, including a human, whose treatment or prevention is affected or promoted by blocking the interaction of menin with MLL proteins and oncogenic MLL fusion proteins.

[0175] The present invention also relates to the use of a compound of formula (I) according to the present invention, or a tautomeric or stereoisomeric form thereof, or a pharma- ceutically acceptable salt or solvate thereof, or a pharmaceutical composition, for the manufacture of a medicament for the treatment, prevention, amelioration, control or reduction of the risk of a disorder associated with the interaction of menin with MLL proteins and oncogenic MLL fusion proteins in a mammal, including a human, whose treatment or prevention is affected or promoted by blocking the interaction of menin with MLL proteins and oncogenic MLL fusion proteins.

[0176] The present invention also relates to a compound of formula (I), a tautomeric or stereoisomeric form thereof, or a pharma- ceutically acceptable salt or solvate thereof, for use in the treatment or prevention of any one of the aforementioned diseases.

[0177] The present invention also relates to a compound of formula (I), a tautomeric or stereoisomeric form thereof, or a pharma- ceutically acceptable salt or solvate thereof, for use in treating or preventing any one of the aforementioned diseases.

[0178] The present invention also relates to the use of a compound of formula (I), a tautomeric or stereoisomeric form thereof, or a pharma- ceutically acceptable salt or solvate thereof, for the manufacture of a medicament for the treatment or prevention of any one of the aforementioned disease conditions.

[0179] The compounds of the invention can be administered to mammals, preferably humans, for the treatment or prevention of any one of the aforementioned diseases.

[0180] In view of the availability of the compounds of formula (I), their tautomeric and stereoisomeric forms, and their pharma- ceutically acceptable salts and solvates, there is provided a method of treating a warm-blooded animal, including a human, suffering from any one of the diseases mentioned hereinbefore.

[0181] The method comprises the administration, i.e. systemic or local, of a therapeutically effective amount of a compound of formula (I), its tautomeric or stereoisomeric forms, or a pharma- ceutically acceptable salt or solvate thereof, to a warm-blooded animal, including man.

[0182] Therefore, the present invention also relates to a method for treating or preventing any one of the aforementioned diseases, comprising administering to a patient in need thereof a therapeutically effective amount of a compound according to the invention.

[0183] Those skilled in the art will recognize that a therapeutically effective amount of a compound of the present invention is an amount sufficient to have therapeutic activity, which amount will vary depending, among other things, on the type of disease, the concentration of the compound in the therapeutic formulation, and the condition of the patient. An effective therapeutic daily amount can be about 0.005 mg / kg to 100 mg / kg. The amount of a compound according to the present invention, also referred to herein as the active ingredient, required to achieve a therapeutic effect can vary on an individual basis, depending, for example, on the particular compound, the route of administration, the age and condition of the recipient, and the particular disorder or disease being treated. The treatment method can also include administering the active ingredient in a regimen of 1 to 4 intakes per day. In these treatment methods, the compound according to the present invention is preferably formulated prior to administration.

[0184] The present invention also provides a composition for preventing or treating the disorders mentioned herein, said composition comprising a therapeutically effective amount of a compound of formula (I), a tautomeric or stereoisomeric form thereof, or a pharma- ceutically acceptable salt or solvate thereof, and a pharma- ceutically acceptable carrier or diluent.

[0185] While it is possible for the active ingredient (e.g., a compound of the present invention) to be administered alone, it is preferable to administer it as a pharmaceutical composition. Thus, the present invention further provides a pharmaceutical composition comprising a compound according to the present invention together with a pharma- ceutically acceptable carrier or diluent. The carrier or diluent must be "acceptable" in the sense of being compatible with the other ingredients of the composition and not deleterious to the recipient thereof.

[0186] Pharmaceutical compositions can be prepared, for example, as described in Gennaro et al., Remington's Pharmaceutical Sciences (18 th They may be prepared by any method well known in the art of pharmacy, such as those described in "Pharmaceutical preparations and their Manufacture," ed., Mack Publishing Company, 1990, especially Part 8: Pharmaceutical preparations and their Manufacture.

[0187] The compounds of the invention can be used alone or in combination with one or more additional therapeutic agents. Combination therapy includes administering a single pharmaceutical dosage formulation containing a compound according to the invention and one or more additional therapeutic agents, as well as administering a compound according to the invention and each additional therapeutic agent in its own separate pharmaceutical dosage formulation.

[0188] Thus, an embodiment of the present invention relates to a product comprising a compound according to the invention as a first active ingredient and one or more anti-cancer agents as further active ingredients as a combined preparation for simultaneous, separate or sequential use in the treatment of patients suffering from cancer.

[0189] One or more other pharmaceutical agents and the compound according to the present invention may be administered simultaneously (e.g., in separate or single compositions) or sequentially in any order. In the latter case, the two or more compounds are administered within a period and in an amount and manner sufficient to ensure that an advantageous or synergistic effect is achieved. It will be understood that the preferred method and order of administration and the respective dosages and regimens for each component of the combination will depend on the specific other pharmaceutical agents and compounds of the present invention administered, their administration routes, the specific condition, particularly the specific tumor being treated, and the specific host being treated.

[0190] The following examples further illustrate the invention. EXAMPLES

[0191] Several methods for preparing the compounds of the present invention are illustrated in the following examples. Unless otherwise specified, all starting materials were obtained from commercial suppliers and used without further purification, or alternatively, could be synthesized by one of ordinary skill in the art by using well-known methods.

[0192] [Table 1-1]

[0193] [Table 1-2]

[0194] [Table 1-3]

[0195] As will be understood by those skilled in the art, compounds synthesized using the protocols shown may exist as solvates, e.g., hydrates, and / or may contain residual solvents or small amounts of impurities. Compounds or intermediates isolated as salt forms may be of integer stoichiometry, i.e., mono- or di-salts, or intermediate stoichiometry. When intermediates or compounds in the experimental part below are shown as "HCl salts" without indicating the number of equivalents of HCl, this means that the number of equivalents of HCl was not determined. The same principle applies to all other salt forms mentioned in the experimental part, e.g.

[0196] [ka] This also applies to.

[0197] The stereochemical configuration of centers in some compounds may be designated as "R" or "S" when the mixtures are separated, and the compounds themselves have been isolated as single stereoisomers and are enantiomerically pure, but for some compounds the stereochemical configuration at the indicated centers may be designated as "R" or "S" if the absolute stereochemistry has not been determined (even if the bonds are drawn stereospecifically). * R" (if the column conditions for separation are described in the synthesis protocol and if only one stereocenter is present or indicated, it will be eluted first from the column) or " * S" (second to elute from the column if the column conditions for separation are described in the synthetic protocol and if only one stereocenter is present or indicated).

[0198] For example, compound 11

[0199] [ka] It should be clear that:

[0200] [ka]

[0201] The stereochemical configuration of the two stereocenters is * (for example, * R or * For compounds represented by (S), the compound itself has been isolated as a single stereoisomer and is enantiomerically pure, but the absolute stereochemistry of the stereocenter has not been determined (even if the bonds are drawn stereospecifically). In this case, the configuration of the first stereocenter is independent of the configuration of the second stereocenter in the same compound. * R" or " * S” is randomly assigned to such molecules.

[0202] For example, in the case of compound 24,

[0203] [ka] This means that the compound is:

[0204] [ka]

[0205] Those skilled in the art will appreciate that the above paragraphs regarding stereochemical configuration also apply to intermediates.

[0206] Those skilled in the art will understand that, even if not explicitly mentioned in the experimental protocols below, typically after column chromatography purification, the desired fractions were collected and the solvent was evaporated.

[0207] When stereochemistry is not indicated, this is meant to be a mixture of stereoisomers unless otherwise indicated or clear from the context.

[0208] When a stereocenter is designated "RS," this means that a racemic mixture was obtained at the indicated center, unless otherwise indicated.

[0209] Preparation of intermediates For intermediates that were used in the next reaction step as crude intermediates or as partially purified intermediates, in some cases either no molar amount is mentioned for such intermediate in the next reaction step or an estimated or theoretical molar amount is indicated for such intermediate in the next reaction step in the reaction protocols described below.

[0210] Preparation of intermediate 1:

[0211] [ka]

[0212] To a mixture of 5-fluoro-2-methoxybenzoic acid (8.00 g, 47.0 mmol) and N-ethylpropan-2-amine (8.19 g, 94.0 mmol) in dry DCM (150 mL) cooled at 0 °C, HATU (21.5 g, 56.5 mmol) and DIEA (9.10 g, 70.4 mmol) were slowly added in portions. The resulting mixture was slowly warmed to room temperature and stirred for 8 h. The organic layer was washed with water (20 mL × 3) and dried over anhydrous Na2SO4. After filtration, the solvent was removed under reduced pressure and the crude product was purified by FCC (EtOAc / PE = 0%-20%) to give the title intermediate 1 (12.0 g, 96% yield) as a white solid.

[0213] The following intermediates were synthesized by methods similar to those described above for Intermediate 1.

[0214] [Table 2]

[0215] Preparation of intermediate 3:

[0216] [ka]

[0217] To a solution of intermediate 1 (12.0 g, 50.1 mmol) in dry DCM (100 mL) cooled to -78 °C, BBr3 (14.4 mL, 152 mmol) was added slowly, and the resulting mixture was slowly warmed to room temperature and stirred for 8 h. The mixture was cooled to -78 °C again, and MeOH (5 mL) was added dropwise to quench the reaction. The resulting mixture was slowly warmed to room temperature, and the pH value was adjusted to about 8 by adding saturated aqueous NaHCO3 solution. The aqueous layer was extracted with DCM (50 mL x 3), and the combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the crude product, which was purified by FCC (EtOAc / PE = 0%-20%) to give the title intermediate 2 (9.0 g, 78% yield) as a white solid.

[0218] The following intermediates were synthesized by methods similar to those described above for intermediate 3.

[0219] [Table 3]

[0220] Preparation of intermediate 5:

[0221] [ka]

[0222] To a solution of 3,5,6-trichloro-1,2,4-triazine (10.0 g, 54.2 mmol) and TEA (15.2 mL, 109 mmol) in DCM (100 mL) cooled at 0 °C, tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate (9.21 g, 43.4 mmol) was added, and the mixture was warmed to room temperature and stirred for 1 h. The mixture was diluted with water (20 mL) and extracted with DCM (30 mL × 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to give the crude product, which was purified by FCC on silica gel (PE / EtOAc = 1:0 to 3:1) to give the title intermediate 3 (12.0 g, 58% yield) as a yellow solid.

[0223] Preparation of intermediate 6:

[0224] [ka]

[0225] A mixture of intermediate 5 (12.0 g, 33.3 mmol), intermediate 2 (7.5 g, 33.3 mmol) and DBU (6.1 g, 40.1 mmol) in THF (120 mL) was stirred at 25 °C for 8 h. The mixture was diluted with water (30 mL) and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was purified by FCC (PE / EtOAc = 1:0 to 3:1) to give the title intermediate 4 (14.0 g, 73% yield) as a green solid.

[0226] The following intermediates were synthesized by methods similar to those described above for intermediate 6.

[0227] [Table 4]

[0228] Preparation of intermediate 8:

[0229] [ka]

[0230] To a mixture of intermediate 6 (20 g, 36.4 mmol), NaBH4 (2.48 g, 65.7 mmol) and TMEDA (8.54 g, 73.5 mmol) in THF (500 mL) was added Pd(dppf)Cl2·DCM (1.70 g, 2.08 mmol) under N2 atmosphere. After addition, the reaction mixture was stirred at 25 °C for 14 h. The reaction mixture was filtered, the filtrate was concentrated and the residue was purified by FCC on silica gel (EtOAc) to give the title intermediate 5 (15 g, 93% purity, 74% yield) as a brown solid.

[0231] The following intermediates were synthesized by methods similar to those described above for Intermediate 8.

[0232] [Table 5]

[0233] Preparation of intermediate 10:

[0234] [ka]

[0235] To a solution of intermediate 8 (300 mg, 0.583 mmol) in DCM (5 mL) was added TFA (0.5 mL, 6.4 mmol), and the resulting mixture was stirred at room temperature for 3 h. Then 10% NaOH (5 mL) solution was added slowly to the mixture to adjust the pH value to about 12, and the resulting mixture was extracted with DCM (10 mL x 3). The combined organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give the title intermediate 6 (220 mg, 90% yield) as a white solid.

[0236] The following intermediates were synthesized by methods similar to those described above for intermediate 10.

[0237] [Table 6]

[0238] Preparation of intermediate 12:

[0239] [ka]

[0240] To a solution of Mg (7.15 g, 294 mmol) and I2 (0.105 g, 0.414 mmol) in THF (176 ml) at 50° C. under N2. The reaction mixture was added dropwise to a solution (10 ml) of 2-(2-bromoethyl)-1,3-dioxolane (25.0 g, 138 mmol) in THF (100 ml). The color of the mixture changed from brown to colorless and the color returned to room temperature. The reaction mixture was then added dropwise over 1 h to a solution (90 ml) of 2-(2-bromoethyl)-1,3-dioxolane (25.0 g, 138 mmol) in THF (100 ml) at 25° C. Finally, the crude product was added dropwise to a solution of N-methoxy-N-methylisobutyramide (11.9 g, 90.7 mmol) in THF (100 ml) at 25 °C under N2 for 16 h. TLC (PE / EA = 4 / 1, Rf = 0.3) showed that a new spot was found. Then 300 mL of saturated aqueous NH4Cl was added to the mixture to quench the reaction. The mixture was warmed to room temperature and filtered. The filtrate was extracted with EA (300 mL x 3), dried over Na2SO4, and filtered. The filtrate was evaporated to dryness, which was purified by FCC (PE / EA = 4 / 1) to give intermediate 13 (22.0 g, 80% purity) as a colorless oil.

[0241] The following intermediates were synthesized by methods similar to those described above for Intermediate 12.

[0242] [Table 7]

[0243] Preparation of intermediates 14, 14a and 14b:

[0244] [ka]

[0245] A stir bar, intermediate 10 (1.5 g, 3.62 mmol), intermediate 12 (2.7 g, 15.7 mmol), acetic acid (500 mg, 8.33 mmol) and methanol (20 mL) were added to a 100 mL round bottom flask and the mixture was heated and stirred at 45° C. for 1 h, after which sodium cyanoborohydride (500 mg, 7.96 mmol) was added to the mixture and the mixture was heated and stirred at 45° C. for 8 h. The reaction mixture was cooled to room temperature and diluted with dichloromethane (20 mL). Subsequently, a saturated solution of sodium bicarbonate (40 mL) was added and the mixture was extracted with dichloromethane (20 mL×3). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue, which was purified by FCC (eluent: dichloromethane:methanol=1:0 to 10:1, dichloromethane:methanol=10:1, Rf=0.4) to give intermediate 14 (1.5 g, purity 96.81%, yield 70.31%) as a colorless oil.

[0246] Intermediate 14 was separated by SFC (separation conditions: DAICEL CHIRALPAK IG (250 mm * 50 mm, 10 um), mobile phase: A: supercritical CO2, B: 0.1% NH3H2O ​​MEOH, A:B=55:45, 200 mL / min, column temperature: 38, nozzle pressure: 100 bar, nozzle temperature: 60, evaporator temperature: 20, trimmer temperature: 25, wavelength: 220 nm). Pure fractions were collected and the solvent was evaporated under vacuum to give two residues. The first residue was partitioned between acetonitrile (2 mL) and water (10 mL). The solution was lyophilized to dryness to give intermediate 14a (2.0 g, 99.33% purity, 39.73% yield) as a white solid. The second residue was partitioned between acetonitrile (2 mL) and water (10 mL). The mixture was lyophilized to dryness to give intermediate 14b (2.0 g, 99.79% purity, 39.92% yield) as a white solid.

[0247] The following intermediates were synthesized by methods similar to those described above for intermediate 14.

[0248] [Table 8]

[0249] Preparation of intermediate 16:

[0250] [ka]

[0251] After adding a stir bar, intermediate 14a, hydrochloric acid (1M, 4.4 mL) and acetonitrile (20 mL) to a 100 mL round bottom flask, the mixture was heated and stirred at 50° C. for 1 h. The mixture was cooled to room temperature, suspended in dichloromethane (40 mL) and adjusted to pH=12 with a 10% solution of sodium hydroxide (10 mL). The aqueous layer was extracted with dichloromethane (10 ml×3). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give intermediate 16 (460 mg, crude) as a pale yellow solid.

[0252] The following intermediates were synthesized by methods similar to those described above for intermediate 16.

[0253] [Table 9]

[0254] Preparation of intermediate 18:

[0255] [ka]

[0256] To a mixture of intermediate 16 (200 mg, 0.380 mmol) in MeOH (3 mL) was added tert-butyl (2-(2-aminoethoxy)ethyl)carbamate (78 mg, 0.382 mmol) and acetic acid (50 mg, 0.833 mmol). The mixture was stirred at room temperature for 30 min. Then NaBH3CN (50 mg, 0.796 mmol) was added to the mixture and the resulting mixture was stirred at room temperature overnight. The reaction mixture was evaporated to remove the solvent. The residue was diluted with saturated aqueous NaHCO3 (10 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give the crude, which was purified by FCC (100% dichloromethane to DCM / MeOH=10 / 1, TLC: dichloromethane:MeOH=10:1, Rf=0.3) to give intermediate 18 (120 mg, purity 65.86%, yield 29.11%) as a yellow solid.

[0257] Preparation of intermediate 19:

[0258] [ka]

[0259] To a mixture of intermediate 18 (110 mg, 0.154 mmol) in MeOH (3 mL) was added formaldehyde (253 mg, 37% solution in H2O, 3.12 mmol) and acetic acid (20 mg, 0.333 mmol). The mixture was stirred at room temperature for 30 min. Then, NaBH3CN (20 mg, 0.318 mmol) was added to the mixture and the resulting mixture was stirred at room temperature overnight. The reaction mixture was evaporated to remove the solvent. The residue was diluted with saturated aqueous NaHCO3 (10 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (column: Xtimate C18 100 * 30mm *The mixture was purified using a 3 μm column, mobile phase A: water (0.225% FA), mobile phase B: acetonitrile, flow rate 25 mL / min, gradient conditions 10% B to 40% B. Pure fractions were collected and the volatile solvents were evaporated under vacuum to give a residue, which was lyophilized to give intermediate 19 (45 mg, purity 97.89%, yield 39.28%) as a yellow oil.

[0260] Preparation of intermediate 20:

[0261] [ka]

[0262] NaH (423 mg, 10.6 mmol) and 10 mL of THF were added to a 100 mL three-neck flask, then tert-butyl (4-hydroxybutyl)carbamate (1 g, 5.28 mmol) in 5 mL of THF was added under argon atmosphere. The mixture was stirred at 0-5 °C for 30 min under argon atmosphere. Subsequently, ethyl 2-bromoacetate (1.32 g, 7.93 mmol) in 5 mL of THF was added to the above solution. The mixture was stirred at room temperature for 16 h. The reaction mixture was poured into 40 mL of water and extracted with EtOAc (40 mL x 2), the combined extracts were washed with brine (50 mL), dried over Na2SO4, filtered, concentrated in vacuo, and the residue was purified by column chromatography (PE / EtOAc = 10-30%) to give intermediate 20 (500 mg, 31% yield) as a colorless oil.

[0263] Preparation of intermediate 21:

[0264] [ka]

[0265] To a solution of intermediate 20 (300 mg, 0.926 mmol) in 10 mL of toluene was added DIBAL-H (1.7 mL, 1M) dropwise under N2 atmosphere at below -68 °C. After the addition, the reaction mixture was stirred at -70 °C for 1 h. 2 mL of MeOH was added to the reaction mixture below -68 °C, then the reaction mixture was added to 20 mL of 0.1 N HCl, extracted with EtOAc (20 mL x 2), the combined extracts were washed with brine (20 mL), dried over Na2SO4, filtered and concentrated in vacuo to give intermediate 21 (170 mg, crude) as a light brown oil, which was used directly in the next step.

[0266] Preparation of intermediate 22:

[0267] [ka]

[0268] To a solution of 4-(methylamino)butanoic acid (10.0 g, 65.1 mmol) and TEA (26.0 mL, 196 mmol) in MeOH (120 mL) was added Boc2O (16.0 g, 73.3 mmol) dropwise. The mixture was stirred at room temperature for 2 days. The mixture was concentrated under reduced pressure to give a residue. The residue was diluted with EtOAc (150 mL), washed with cold 0.1 N HCl (70 mL x 2), then with H2O (50 mL x 2) and brine (50 mL), dried over Na2SO4, filtered and concentrated to give the desired product (8.90 g, crude) as a colorless oil. The crude product was used in the next step without further purification.

[0269] Preparation of intermediate 23:

[0270] [ka]

[0271] A stir bar, intermediate 22 (8.90 g, 41.0 mmol), N,O-dimethylhydroxylamine hydrochloride (5.00 g, 51.3 mmol), 1H-benzo[d][1,2,3]triazol-1-ol (5.50 g, 40.7 mmol), 4-methylmorpholine (25.0 g, 247 mmol) and CHCl3 (300 mL) were added to a 1 L round bottom flask under N2 atmosphere. The reaction mixture was cooled to 0 °C and N1-((ethylimino)methylene)-N3,N3-dimethylpropane-1,3-diamine hydrochloride (11.0 g, 57.4 mmol) was added. The reaction mixture was then stirred at 27 °C for 16 h. The reaction mixture was washed with water (200 mL), then with 0.1 N HCl (150 mL×2), saturated NaHCO3 (200 mL×3), brine (200 mL), dried over Na2SO4, filtered, concentrated in vacuo, and the residue was purified by column chromatography (PE / EtOAc=20%-40%) to give intermediate 23 (6.30 g, 59% yield) as a colorless oil.

[0272] Preparation of intermediate 24:

[0273] [ka]

[0274] Isopropyllithium (64.8 mL, 45.4 mmol, 0.7 M) was added dropwise to a solution of intermediate 23 (4.00 g, 15.4 mmol) in THF (50 mL) at -70 °C under N2. The solution was stirred at 70 °C for 2 h. The mixture was quenched with saturated NH4Cl solution (80 mL) and extracted with EtOAc (50 mL x 3). The combined organic phase was washed with brine (50 mL), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by FCC (PE: EtOAc = 10: 1) to give intermediate 24 (2.20 g, 55% yield) as a colorless oil.

[0275] Preparation of intermediates 25, 25a and 25b:

[0276] [ka]

[0277] To a solution of intermediate 10 (1.2 g, 2.90 mmol) and intermediate 24 (705 mg, 2.90 mmol) in MeOH (50 mL) was added ZnCl2 (1.60 g, 11.7 mmol). The mixture was stirred at 80 °C for 2 h. Then sodium cyanotrihydroborate (1.1 g, 17.5 mmol) was added. The reaction mixture was stirred at 80 °C overnight. The mixture was concentrated under reduced pressure to give a residue. The residue was diluted with DCM (100 mL) and quenched with saturated NH4Cl (50 mL). The aqueous was extracted with DCM (50 mL x 3). The combined organic phase was washed with brine (50 mL), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by FCC (DCM:MeOH=10:1) to give intermediate 25 (1.0 g, 48% yield) as a white solid, which was resolved by SFC to give intermediate 25a (400 mg, 40% yield) (peak 1, Rt=1.326 min) and intermediate 25b (400 mg, 40% yield) (peak 2, Rt=1.420 min), all as white solids. SFC method: Column: DAICEL CHIRALPAK AD (250 mm * 50mm, 10um), Mobile phase: A: Supercritical CO2, B: 0.1% NH3H2O ​​IPA, A:B=75:25, 200mL / min, Column temperature: 38℃, Nozzle pressure: 100 bar, Nozzle temperature: 60℃, Evaporator temperature=20℃, Trimmer temperature: 25℃, Wavelength: 220nm.

[0278] Preparation of intermediate 26:

[0279] [ka]

[0280] To a solution of intermediate 25a (100 mg, 0.156 mmol) in 2 mL of dioxane was added HCl / dioxane (4 mL, 16 mmol). After the addition, the reaction mixture was stirred at room temperature for 1 h. The reaction mixture was concentrated in vacuo to give intermediate 26 (92 mg, crude) as a colorless oil, which was used in the next step without purification.

[0281] Preparation of intermediate 27:

[0282] [ka]

[0283] To a solution of intermediate 26 (220 mg, 0.358 mmol) and intermediate 21 (170 mg, 0.735 mmol) in 15 mL of MeOH was added NaOAc (100 mg, 1.22 mmol). After stirring for 15 min, NaBH3CN (50 mg, 0.796 mmol) was added to the mixture. After the addition, the reaction mixture was stirred and heated at 35° C. for 16 h. The reaction mixture was concentrated in vacuo and the residue was diluted with 30 mL of water and basified to pH=12 by 5% NaOH, then extracted with DCM (30 ml×3). The combined extracts were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered, concentrated in vacuo, and the residue was purified by column chromatography (eluent: 100% DCM to DCM / MeOH=20:1 (containing 0.25% NH3.H2O)) to give intermediate 27 (210 mg, 72% yield) as a light brown sticky oil.

[0284] Preparation of intermediate 28:

[0285] [ka]

[0286] To a solution of tert-butyl (2-aminoethyl)carbamate (5.00 g, 31.2 mmol) and propan-2-one (3.62 g, 62.3 mmol) in MeOH (50 mL) was added acetic acid (3.81 g, 62.4 mmol). The mixture was stirred at room temperature for 0.5 h, then sodium cyanotrihydroborate (3.93 g, 62.5 mmol) was added. The mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure, and the mixture was diluted with DCM (300 mL). The mixture was washed with NaHCO3 (100 mL x 2) and brine (100 mL x 2), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel (eluent: 100% dichloromethane to dichloromethane:methanol=10:1) to give intermediate 28 (5.0 g, 79% yield) as a yellow oil.

[0287] Preparation of intermediate 29:

[0288] [ka]

[0289] To a solution of 5-fluoro-2-methoxybenzoic acid (2.50 g, 14.1 mmol) and intermediate 28 (2.86 g, 14.1 mmol) in DCM (50 mL) was added T3P (18.0 g, 28.3 mmol) and TEA (6.25 mL, 45.0 mmol). The mixture was stirred at room temperature for 1 h. The mixture was diluted with DCM (100 mL) and the mixture was washed with NaHCO3 (50 mL x 2) and brine (50 mL x 2), dried over Na2SO4, filtered and concentrated under reduced pressure to give intermediate 29 (5.0 g, crude) as a yellow oil, which was used in the next step without further purification.

[0290] Preparation of intermediate 30:

[0291] [ka]

[0292] To a solution of intermediate 29 (5.00 g, 14.1 mmol) in DCM (50 mL) was added BBr3 (4.00 mL, 42.3 mmol) at -78 °C under N2 atmosphere. The mixture was stirred at -78 °C for 1 h and at room temperature for 12 h. The mixture was quenched with H2O (20 mL) at 0 °C. The mixture was extracted with DCM (100 mL), the organic phase was discarded and the aqueous phase was basified with NaOH (2 M) to pH = 11 to give a solution (3.40 g in solution, theoretical amount) which was used in the next step without further purification.

[0293] Preparation of intermediate 31:

[0294] [ka]

[0295] To a solution of intermediate 30 (3.40 g, 14.2 mmol, pH=11 in HO (50 mL)) was added BocO (3.09 g, 14.2 mmol). The mixture was stirred at room temperature for 1 h. The mixture was diluted with water (50 mL), extracted with DCM (30 mL×3), dried over NaSO, filtered and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel (eluent: 100% petroleum ether to petroleum ether:ethyl acetate=1:1) to give intermediate 31 (900 mg, purity 90.75%, yield 17.0%) as a white solid.

[0296] Preparation of intermediate 32:

[0297] [ka]

[0298] To a solution of intermediate 31 (900 mg, 2.64 mmol) in DCM (20 mL) was added ethyl 6-oxo-1,6-dihydro-1,2,4-triazine-5-carboxylate (894 mg, 5.29 mmol), TEA (1.30 mL, 13.1 mmol) and PyBrop (2.47 g, 5.30 mmol). The mixture was stirred at 30 °C for 12 h. The mixture was diluted with DCM (30 mL), washed with water (20 mL x 2), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel (eluent: A: petroleum ether, B: ethyl acetate, 0% B to 50% B in A) to give intermediate 32 (650 mg, 94.50% purity, 47% yield) as a brown solid.

[0299] Preparation of intermediate 33:

[0300] [ka]

[0301] To a solution of intermediate 21 (340 mg, 0.692 mmol) in THF (6 mL) and HO (1 mL) was added LiOH·HO (44.0 mg, 1.05 mmol) at 0 °C. After the addition, the reaction mixture was stirred at 20 °C for 0.5 h. The mixture was adjusted to pH = 5 with 1N HCl and evaporated under reduced pressure to remove the solvent. The residue was lyophilized to give intermediate 33 (320 mg, crude) as a yellow solid, which was used in the next step without further purification.

[0302] Preparation of intermediate 34:

[0303] [ka]

[0304] 4A molecular sieves (1.0 g) was added to a solution of intermediate 33 (320 mg, 0.690 mmol) in 2,2,2-trifluoroethanol (5 mL). The mixture was stirred at 70 °C for 1 h. Dibromoisocyanuric acid (396 mg, 1.38 mmol) was then added. The mixture was stirred at 70 °C for 1 h. The resultant was filtered. The filtrate was concentrated and purified by FCC (eluent: A: petroleum ether, B: ethyl acetate, 0% B to 50% B in A) to give intermediate 34 (170 mg, 45% yield) as a yellow oil.

[0305] Preparation of intermediate 35:

[0306] [ka]

[0307] Intermediate 34 (170 mg, 0.329 mmol), ( * SN-(2-Methoxyethyl)-N,5-dimethyl-4-(2,6-diazaspiro[3.4]octan-2-yl)hexan-1-amine hydrochloride (132 mg, 0.395 mmol), DBU (0.15 mL, 1.00 mmol), and CH3CN (5 mL) were added to a 100 mL round-bottom flask. The mixture was stirred at 25 °C for 1 h. The mixture was concentrated under reduced pressure to remove the solvent, diluted with DCM (10 mL), and washed with H2O (10 mL) and brine (10 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (column: Phenomenex Gemini-NX 80 * 40mm * The mixture was purified using 3 um, mobile phase A: water (0.05% NH3H2O ​​+ 10 mM NH4HCO3), mobile phase B: acetonitrile, flow rate: 30 mL / min, gradient conditions 44%B to 74%). Pure fractions were collected and volatile solvents were evaporated under vacuum. The resulting aqueous mixture was extracted with DCM (20 mL x 2). The combined organic extracts were dried over Na2SO4, filtered and concentrated to dryness under reduced pressure to give intermediate 35 (170 mg, 72% yield) as a yellow oil.

[0308] Preparation of intermediate 37:

[0309] [ka]

[0310] To a solution of intermediate 36 (400 mg, 0.871 mmol) in xylene (2 mL), 1-(ethylamino)propan-2-ol (449 mg, 4.353 mmol) was added and stirred at 135° C. for 20 h. The mixture was then concentrated under reduced pressure to give a residue which was purified by column chromatography on silica gel (10% MeOH in CH2Cl2) to give intermediate 37 as an oil (462 mg, 90% purity, 36% yield).

[0311] Preparation of intermediate 38:

[0312] [ka]

[0313] To a solution of intermediate 37 (200 mg, 0.377 mmol) in methylene chloride (2.1 mL, 1.326 g / mL, 32.786 mmol) was added TFA (0.7 mL, 1.49 g / mL, 9.147 mmol) dropwise at room temperature and then stirred at room temperature for 1 h. The mixture was then concentrated under reduced pressure to give intermediate 38, which was used directly in the next step.

[0314] Preparation of intermediate 39:

[0315] [ka]

[0316] To a solution of benzyl 2,6-diazaspiro[3.4]octane-6-carboxylate (5.0 g, 20.3 mmol) in 100 mL of MeOH was added HCl / dioxane (5.1 mL, 20.4 mmol, 4 M). The mixture was stirred at 30° C. for 30 min. Then AcONa (5.0 g, 61.0 mmol) and 6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-one (6.40 g, 31.8 mmol) were added. The reaction mixture was stirred at 30° C. for 15 min and NaBH3CN (2.0 g, 31.8 mmol) was added. After the addition, the reaction mixture was stirred at 30° C. for 15 h. The reaction mixture was concentrated and the residue was diluted with DCM (100 mL) and washed with 2N HCl (200 mL×2). The combined aqueous was washed with DCM (100 mL), then basified to pH=12 with 10% NaOH and extracted with DCM (200 mL×3). The combined extracts were washed with brine (200 mL), dried over Na2SO4, filtered and concentrated to give the crude compound. The crude product was purified by FCC (DCM:MeOH=10:1) to give intermediate 39 (2.60 g, 30% yield) as a colorless liquid.

[0317] Preparation of intermediate 40:

[0318] [ka]

[0319] Intermediate 39 (2.0 g, 4.63 mmol) was purified by supercritical fluid chromatography (column: DAICEL CHIRALPAK AY-H (250 mm * The mixture was purified using a 30 mm column (30 mm, 5 um), mobile phase A: 0.1% NH3H2O, mobile phase B: EtOH, flow rate: 100 mL / min, gradient conditions 20% B to 20%). The first fraction was collected and evaporated to give intermediate 40 (Rt = 2.309 min, 590 mg, 30% yield) as a colorless oil.

[0320] Preparation of intermediate 41:

[0321] [ka]

[0322] After adding a stir bar, intermediate 40 (690 mg, 1.45 mmol), 1,1,2-trichloroethane (964 mg, 7.22 mmol) and anhydrous methanol (20 mL) to a 100 mL hydrogenation bottle, the mixture was purged with argon three times, and then wet Pd / C (120 mg, 10% purity) was added to the mixture. The resulting mixture was purged with argon and hydrogen three times, and heated and stirred under hydrogen atmosphere (50 psi) at 40° C. for 4 hours. The mixture was cooled to room temperature, filtered through a pad of Celite®, and the filter cake was washed with methanol (10 mL×3). The combined organic layers were concentrated under reduced pressure to give intermediate 41 (120 mg, crude) as a colorless oil.

[0323] Preparation of intermediate 42:

[0324] [ka]

[0325] A stir bar, 3,5,6-trichloro-1,2,4-triazine (1.00 g, 5.42 mmol), intermediate 41 (1.61 g, 5.41 mmol) and anhydrous dichloromethane (20 mL) were added to a 40 mL glass bottle, and then triethylamine (1.10 g, 10.9 mmol) was added dropwise to the mixture. The resulting mixture was stirred at 25 °C for 8 h. The mixture was concentrated under reduced pressure to give a residue, which was suspended in water (40 mL) and extracted with dichloromethane (20 mL x 3). The combined organic layer was dried over anhydrous Na2SO4. It was purified by FCC (100% dichloromethane to dichloromethane:methanol = 25:1) to give intermediate 42 (1.20 g, purity 96.13%, yield 47.8%) as a yellow solid.

[0326] Preparation of intermediate 43:

[0327] [ka]

[0328] A stir bar, 5-fluoro-2-methoxybenzoic acid (10 g, 58.8 mmol), 2-(isopropylamino)ethanol (12.1 g, 117 mmol), TEA (17.8 g, 176 mmol) and dry dichloromethane (200 mL) were added to a 500 mL round bottom flask, then stirred at 0 °C, after which T3P (56.1 g, 88.2 mmol) was added to the mixture. The resulting mixture was stirred at 25 °C for 12 h. The reaction mixture was poured into dichloromethane (300 mL) and then washed with water (200 mL x 3 times). The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure to give intermediate 43 (11.7 g, 90% purity, 70.2% yield) as a yellow oil.

[0329] Preparation of intermediate 44:

[0330] [ka]

[0331] After adding a stir bar, intermediate 43 (11.7 g, 45.8 mmol) and anhydrous dichloromethane (35 mL) to a 100 mL three-necked round-bottom flask, the mixture was cooled to -78 °C under a dry ice-ethanol bath, and then tribromoborane (9.54 mL, 101 mmol, 2.65 g / mL) was added dropwise to the mixture over 1 h. The resulting mixture was gradually warmed to room temperature and stirred at 25 °C for 1 h. The mixture was cooled to -78 °C under a dry ice-ethanol bath and quenched by dropwise addition of methanol (30 mL). The resulting mixture was slowly added to a saturated solution of sodium bicarbonate (200 mL). The aqueous layer was extracted with dichloromethane (100 mL x 3). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give the crude material which was triturated with petroleum ether / ethyl acetate (50 / 1, 200 mL) to give intermediate 44 (6.56 g, 90% purity, 53.4% ​​yield) as a pale yellow solid.

[0332] Preparation of intermediate 45:

[0333] [ka]

[0334] To a solution of intermediate 42 (524 mg, 1.18 mmol) and intermediate 44 (340 mg, 1.41 mmol) in THF (20 mL) was added DBU (340 mg, 2.23 mmol). The reaction mixture was stirred at 25° C. for 12 h. The reaction mixture was poured into water (30 mL) and extracted with dichloromethane (20 mL×3). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the crude product, which was triturated with petroleum ether / ethyl acetate=6 mL / 0.5 mL, the mixture was filtered, and the filter cake was evaporated under vacuum to give intermediate 45 (371 mg, purity 78.12%, yield 37.89%) as a yellow solid.

[0335] Preparation of intermediates 46, 46a and 46b:

[0336] [ka]

[0337] Intermediate 10 (15 g, 36 mmol) was added to a solution of 1-hydroxy-4-methylpentan-3-one (12.6 g, crude), AcOH (6.2 mL, 108 mmol) and MeOH (200 mL). The mixture was stirred at room temperature for 0.5 h. NaBH3CN (6.8 g, 108 mmol) was added to the mixture. The mixture was then stirred at room temperature for 2 h. The reaction mixture was adjusted to pH=8 with NH3 (7M in MeOH), poured into brine (100 mL) and extracted with ethyl acetate (300 mL×3). The combined organic layers were dried over Na2SO4, filtered and concentrated to dryness under reduced pressure to give the crude product, which was purified by FCC (eluent: petroleum ether:ethyl acetate=1:0-0:1, then ethyl acetate:(dichloromethane:methanol=1:1)=1:0-1:1) to give crude intermediate 46 (17 g) as a white solid.

[0338] Intermediate 46 (1.6 g) was purified by SFC on a DAICEL CHIRALPAK IG 250 mm × 30 mm, 10 μm (isocratic elution: MeOH (containing 0.1% of 25% aqueous NH3):supercritical CO2, 30%:70% to 30%:70% (v / v)). Pure fractions were collected and volatiles removed under reduced pressure. The product was suspended in water (10 mL) and the mixture was frozen using dry ice / acetone, then lyophilized to dryness to give intermediate 46a (1st fraction, 700 mg, 41%) as a white solid and intermediate 46b (2nd fraction, 660 mg, 36%) as a white solid.

[0339] Preparation of intermediate 47:

[0340] [ka]

[0341] MsCl (0.169 mL, 1.48 g / mL, 2.182 mmol) was added dropwise to a solution of intermediate 46a (500 mg, 0.972 mmol), Et3N (0.27 mL, 1.9 mmol) and dichloromethane (10 mL) at 0 °C (ice / water) under N2 atmosphere. The resulting mixture was stirred at 0 °C (ice / water) under N2 for 45 min. It was quenched with water (5 mL) and then extracted with dichloromethane (10 mL x 3). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give intermediate 47 (400 mg, 62.514% yield) as a yellow solid, which was used in the next step without further purification.

[0342] Preparation of intermediate 48:

[0343] [ka]

[0344] Intermediate 47 (200 mg, 0.304 mmol) and 2-aminoethane-1-sulfonamide (264 mg, 2.126 mmol) were added to THF (25 mL, 0.886 g / mL, 307.182 mmol). The mixture was stirred at 60 °C for 16 h. The solvent was removed. The residue was purified by flash column (C18, CHCN:HO 5:95 to 30:70, HCOOH as buffer) to give intermediate 48 (40 mg, 19% yield).

[0345] Preparation of intermediate 49:

[0346] [ka]

[0347] To a solution of N-ethyl-5-fluoro-2-hydroxy-N-isopropylbenzamide (10.0 g, 44.4 mmol) in H2SO4 (20 mL) and trifluoroacetic acid (40 mL) was added NBS (8.7 g, 48.9 mmol). The reaction mixture was stirred at 25°C for 12 h. The reaction mixture was carefully poured onto 200 g of crushed ice. The mixture was extracted with ethyl acetate (100 mL x 3). The combined organic layers were dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by preparative HPLC (column: YMC-Triart Prep C18 250 * 50mm * The mixture was purified using a 10 μm column, mobile phase A: water (0.225% FA), mobile phase B: acetonitrile, flow rate: 100 mL / min, gradient conditions 20% B to 60% B. Pure fractions were collected and the solvent was evaporated to give intermediate 49 (10.0 g, purity 99.48%, yield 73.7%) as a yellow solid.

[0348] Preparation of intermediate 50:

[0349] [ka]

[0350] To a solution of ethyl 6-chloro-1,2,4-triazine-5-carboxylate (650 mg, 3.47 mmol), intermediate 49 (896 mg, 2.95 mmol) in dry DMF (20 mL) was added potassium carbonate (1.44 g, 10.4 mmol). The reaction mixture was stirred at 25° C. for 12 h. The mixture was suspended in water (80 mL) and extracted with ethyl acetate (50 mL×3). The combined organic layers were washed with a saturated solution of lithium chloride (50 mL×3). The aqueous layer was lyophilized to dryness to give intermediate 50 (400 mg, crude) as a yellow solid, which was used in the next step without further purification.

[0351] Preparation of intermediate 51:

[0352] [ka]

[0353] A stir bar, intermediate 50 (300 mg, 0.702 mmol), 4A molecular sieves (1.0 g) and dry 2,2,2-trifluoroethanol (20 mL) were added to a 100 mL round bottom flask. The reaction mixture was heated and stirred at 65° C. for 2 hours under an argon atmosphere. 1,3-dibromo-1,3,5-triazinane-2,4,6-trione (404 mg, 1.41 mmol) was then added to the mixture in one portion. The reaction mixture was cooled to room temperature and stirred at 25° C. for an additional 12 hours. The reaction mixture was filtered and the filter cake was washed with ethyl acetate (50 mL). The filtrate was concentrated under reduced pressure to give a residue, which was purified by flash column chromatography on silica gel (100% petroleum ether to petroleum ether:ethyl acetate=3:1, TLC: petroleum ether:ethyl acetate=3:1, Rf=0.2) to give intermediate 51 (200 mg, purity 94.47%, yield 55.9%) as a red oil.

[0354] Preparation of intermediate 52:

[0355] [ka]

[0356] To a solution of 3,3-dimethylbutan-2-one (5.78 g, 57.7 mmol) in anhydrous tetrahydrofuran (80 mL) was added LDA (28.9 mL, 57.8 mmol, 2M in THF) at −78 °C under N2 atmosphere. After addition, the reaction mixture was stirred at −78 °C for 1 h. Then, tert-butyl methyl (2-oxoethyl) carbamate (5.0 g, 28.9 mmol) in anhydrous tetrahydrofuran (20 mL) was added dropwise to the reaction mixture, and the mixture was stirred at −78 °C for 2 h. The reaction mixture was quenched with aqueous NH4Cl solution (20 mL) at −78 °C, water (100 mL) was added, and then warmed to 25 °C. The mixture was extracted with ethyl acetate (150 mL × 3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (column: Xtimate C18 150 * 40mm * Purification was performed using 5 μm, mobile phase A: water (0.05% NH3H2O), mobile phase B: acetonitrile, flow rate: 60 mL / min, gradient conditions 35% B to 65%). Pure fractions were collected and the solvent was evaporated under vacuum. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The mixture was lyophilized to give intermediate 52 (2.20 g, 100% purity, 27.9% yield) as a colorless oil.

[0357] Preparation of intermediates 53a and 53b:

[0358] [ka]

[0359] ZnCl2 (449 mg, 3.29 mmol) was added to intermediate 11 (704 mg, 1.64 mmol) and intermediate 52 (900 mg, 3.29 mmol) in dry methanol (20 mL). The reaction mixture was heated and stirred at 65° C. for 1 h, then sodium cyanotrihydroborate (207 mg, 3.29 mmol) was added. The reaction mixture was stirred at 65° C. for another 4 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a residue, after which a saturated solution of sodium bicarbonate (80 mL) was added. The mixture was extracted with dichloromethane (50 mL×3). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (column: Xtimate C18 150 * 40mm * The mixture was purified using a 500 mL column at 5 um, mobile phase A: water (0.05% NH3H2O), mobile phase B: acetonitrile, flow rate: 60 mL / min, gradient conditions 50% B to 80%). Pure fractions were collected and the solvent was evaporated under vacuum. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The mixture was lyophilized to give the first fraction (160 mg, purity 96.85%, yield 13.8%) as a colorless sticky oil and the second fraction (150 mg, purity 95.86%, yield 12.8%) as a colorless sticky oil.

[0360] The first fraction (160 mg, 0.233 mmol) was subjected to supercritical fluid chromatography (separation conditions: DAICEL CHIRALPAK IG (250 mm * The separation was carried out using a 100% hexanes water column (30 mm, 10 um), mobile phase: A: supercritical CO2, B: 0.1% NH3H2O ​​ETOH, A:B=65:35, 70 mL / min, column temperature: 38°C, nozzle pressure: 100 bar, nozzle temperature: 60°C, evaporator temperature=20°C, trimmer temperature: 25°C, wavelength: 220 nm). Pure fractions were collected and the solvent was evaporated under vacuum. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The solution was lyophilized to give intermediate 53a (70.0 mg, 100% purity, 43.8% yield) as a colorless sticky oil and intermediate 53b (70.0 mg, 97.24% purity, 42.5% yield) as a colorless sticky oil.

[0361] Preparation of intermediates 54a and 54b:

[0362] [ka]

[0363] To a solution of intermediate 53a (70.0 mg, 0.102 mmol) in anhydrous dichloromethane (2 mL) was added trifluoroacetic acid (2 mL). The reaction mixture was stirred at 25° C. for 30 minutes. The reaction mixture was concentrated under reduced pressure to give intermediate 54a (70.0 mg, crude) as a yellow oil.

[0364] To a solution of intermediate 53b (70.0 mg, 0.102 mmol) in anhydrous dichloromethane (2 mL) was added trifluoroacetic acid (2 mL). The reaction mixture was stirred at 25° C. for 30 minutes. The reaction mixture was concentrated under reduced pressure to give intermediate 54b (70.0 mg, crude) as a yellow oil.

[0365] Preparation of compounds Preparation of Compound 1:

[0366] [ka]

[0367] A stir bar, intermediate 16 (120 mg, 0.228 mmol), N-methyl-2-(methylthio)ethanamine hydrochloride (133 mg, 0.939 mmol) and methanol (3 mL) were added to a 40 mL glass bottle, and the mixture was heated and stirred at 45° C. for 2 h, then sodium cyanoborohydride (70 mg, 1.11 mmol) was added to the mixture, and the mixture was heated and stirred at 45° C. for 8 h. The mixture was cooled to room temperature, and then the mixture was quenched with water (20 mL) and extracted with dichloromethane (30 mL×3), and the organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (column: Boston Prime C18 150 * 30mm *The mixture was purified using 5 μm, mobile phase A: water (0.05% NH3H2O ​​+ 10 mM NH4HCO3), mobile phase B: acetonitrile, flow rate: 25 mL / min, gradient conditions 70% B to 100% B). Pure fractions were collected and the solvent was evaporated under vacuum. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The mixture was lyophilized to dryness to give the product (85 mg, 100% purity, 60.57% yield, free base) as a white solid, which was mixed with fumaric acid (32.2 mg, 0.277 mmol), acetonitrile (12 mL) and water (4 mL) in a 50 mL round bottom flask. The mixture was concentrated under reduced pressure to give a residue. The residue was partitioned between acetonitrile (3 mL) and water (10 mL). The solution was lyophilized to dryness to give compound 1 (81.33 mg, 96.99% purity, 67.40% yield) as a white solid.

[0368] Preparation of Compound 2:

[0369] [ka]

[0370] A stir bar, compound 4 (60 mg, 0.069 mmol, free base form), formaldehyde (55.8 mg, 0.688 mmol), sodium cyanoborohydride (17.2 mg, 0.274 mmol) and methanol (2 mL) were added to an 8 mL glass bottle and the mixture was stirred at 25° C. for 8 h. The mixture was quenched with water (20 mL) and diluted with dichloromethane (20 mL). * 3), and the organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (column: Welch Xtimate C18 150 * 25mm * The pure fractions were purified using a 5 μm column, mobile phase A: water (0.2% FA), mobile phase B: ACN, flow rate: 25 mL / min, gradient conditions 15% B to 45%). The pure fractions were adjusted to pH=8 by adding a 10% solution of sodium hydroxide and diluted with dichloromethane (20 mL *3) and the organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The solution was lyophilized to dryness to give the product (10 mg, 100% purity, 22.23% yield) as a colorless oil, which was mixed with fumaric acid (3.6 mg, 0.031 mmol), acetonitrile (12 mL) and water (4 mL) in a 50 mL round bottom flask. The mixture was concentrated under reduced pressure to give a residue. The residue was partitioned between acetonitrile (3 mL) and water (10 mL). The solution was lyophilized to dryness to give compound 2 (8.8 mg, 94.46% purity, 61.34% yield) as a yellow solid.

[0371] Preparation of compound 3:

[0372] [ka]

[0373] A stir bar, Intermediate 16 (120 mg, 0.228 mmol), 2-(methylthio)ethanamine (80 mg, 0.877 mmol), sodium acetate (110 mg, 1.34 mmol) and methanol (3 mL) were added to an 8 mL glass bottle and the mixture was stirred and heated at 60° C. for 2 hours, after which sodium cyanoborohydride (30 mg, 0.477 mmol) was added to the mixture and the mixture was heated and stirred at 60° C. for 8 hours. The mixture was cooled to room temperature, then water (30 mL) was poured into the mixture and dichloromethane (20 mL) was added. * 3), and the organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give a residue, which was purified by preparative HPLC (column: Boston Prime C18 150 * 30mm *The mixture was purified using 50 mL of 5 um, mobile phase A: water (0.05% NH3H2O ​​+ 10 mM NH4HCO3), mobile phase B: acetonitrile, flow rate: 25 mL / min, gradient conditions 53%B to 83%). Pure fractions were collected and the solvent was evaporated under vacuum. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The mixture was lyophilized to dryness to give the product (36 mg, 99.01% purity, 25.99% yield) as a colorless oil, which was mixed with fumaric acid (14.0 mg, 0.121 mmol), acetonitrile (12 mL) and water (4 mL) in a 50 mL round bottom flask. The mixture was then concentrated under reduced pressure to give a residue. The residue was partitioned between acetonitrile (3 mL) and water (10 mL). The solution was lyophilized to dryness to give compound 3 (36.17 mg, 91.83% purity, 66.58% yield) as a white solid.

[0374] Preparation of compound 4:

[0375] [ka]

[0376] After adding a stir bar, intermediate 16 (200 mg, 0.380 mmol), 2-(trifluoromethoxy)ethanamine hydrochloride (94.3 mg, 0.570 mmol), triethylamine (115 mg, 1.14 mmol) and anhydrous dichloromethane (10 mL) to a 40 mL glass bottle, the resulting mixture was stirred at 25 °C for 1 h, and then sodium triacetoxyborohydride (161 mg, 0.760 mmol) was added to the mixture. The resulting mixture was stirred at 25 °C for another 1 h. The mixture was diluted in dichloromethane (20 mL) and adjusted to pH = 8 by a saturated solution of sodium bicarbonate (10 mL). The aqueous layer was extracted with dichloromethane (10 mL x 2). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (column: Boston Green ODS 150 * 30mm *The mixture was purified with 5 um, mobile phase A: water (0.2% FA), mobile phase B: acetonitrile, flow rate: 35 mL / min, gradient condition 5%B to 35%). Pure fractions were collected and adjusted to pH=12 by a solution of sodium hydroxide (3M, 8 mL). The aqueous layer was extracted with dichloromethane (10 mLx2). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The solution was lyophilized to dryness to give the product (80.0 mg, purity 97.84%, yield 32.2%) as a colorless solid, which was mixed with fumaric acid (29.0 mg, 0.250 mmol) and acetonitrile (5 mL) in a 50 mL round bottom flask, and the resulting mixture was stirred at 25°C for 1 h. The resulting mixture was concentrated under reduced pressure to give a residue. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The solution was lyophilized to dryness to give compound 4 (91.20 mg, 89.68% purity, 75.0% yield) as a white powder.

[0377] Preparation of compound 5:

[0378] [ka]

[0379] To a mixture of intermediate 19 (45 mg, 0.062 mmol) in DCM (3 mL) was added TFA (1 mL) at 0° C. The reaction mixture was stirred at room temperature for 1 h. The reaction mixture was evaporated under reduced pressure to remove the solvent. The residue was diluted with saturated aqueous NaHCO3 (5 mL) and CHCl2 (5 mL) and basified with NaOH (2N) to pH=12. The resulting mixture was stirred at room temperature for 30 min. The mixture was separated and the aqueous layer was extracted with dichloromethane (5 mL×2). The combined organic layers were washed with brine (5 mL), dried over anhydrous NaSO4, filtered and concentrated under reduced pressure to give the product (36 mg, 92.74% yield) as a yellow oil, which was mixed with fumaric acid (20 mg, 0.172 mmol), acetonitrile (12 mL) and methanol (4 mL) in a 50 mL round bottom flask. The mixture was then concentrated under reduced pressure to give a residue. The residue was partitioned between acetonitrile (3 mL) and water (10 mL). The solution was lyophilized to dryness to give compound 5 (31.06 mg, 91.37% purity, 50.74% yield) as a yellow powder.

[0380] Preparation of compound 6:

[0381] [ka]

[0382] To a solution of intermediate 27 (210 mg, 0.277 mmol) in 3 mL of MeOH was added 5 mL of HCl / dioxane (4 M). After addition, the reaction mixture was stirred at 10° C. for 30 min. The reaction mixture was concentrated in vacuo and the residue (201 mg) was purified by preparative HPLC (column Venusil ASB Phenyl 150 * 30mm * The mixture was purified using a 5 μm column, mobile phase A: water (0.05% HCl), mobile phase B: acetonitrile, flow rate: 30 mL / min, gradient conditions: 5% B to 35%). Pure fractions were collected and lyophilized to give compound 6 (130 mg, 60% yield) as a white solid.

[0383] Preparation of compound 7:

[0384] [ka]

[0385] A stir bar, Intermediate 16 (200 mg, 0.380 mmol), 2-(difluoromethoxy)ethanamine hydrochloride (112 mg, 0.759 mmol), triethylamine (384 mg, 3.80 mmol) and dichloromethane (2 mL) were added to an 8 mL round bottom flask and the mixture was stirred at 25° C. for 2 hours, after which sodium triacetoxyborohydride (241 mg, 1.14 mmol) was added to the mixture and the mixture was stirred at 25° C. for 8 hours. The mixture was quenched with water (20 mL) and diluted with dichloromethane (30 mL). * 3), and the organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (column: Welch Xtimate C18 150 * 25mm * The mixture was purified using a 5 μm column, mobile phase A: water (0.2% FA), mobile phase B: acetonitrile, flow rate: 25 mL / min, gradient conditions 5% B to 35%). Pure fractions were collected and the solvent was evaporated under vacuum. The residue was adjusted to pH = 8 with a solution of 10% sodium hydroxide (20 mL) and diluted with dichloromethane (30 mL). * 3), the organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue, which was partitioned between acetonitrile (2 mL) and water (10 mL). The mixture was lyophilized to dryness to give the product (90 mg, 98.90% purity, 37.70% yield) as a colorless oil, which was mixed with fumaric acid (8.6 mg, 0.074 mmol), acetonitrile (12 mL) and water (4 mL) in a 50 mL round-bottom flask. The mixture was concentrated under reduced pressure to give a residue. The residue was partitioned between acetonitrile (3 mL) and water (10 mL). The solution was lyophilized to dryness to give compound 7 (21.45 mg, 92.07% purity, 62.52% yield) as a white solid.

[0386] Preparation of compound 8:

[0387] [ka]

[0388] A stir bar, compound 7 (60 mg, 0.097 mmol, free base form), formaldehyde (78.0 mg, 37% solution in HO, 0.961 mmol) and methanol (2 mL) were added to a 50 mL round bottom flask and the mixture was stirred at 25° C. for 2 h, after which sodium cyanoborohydride (24.2 mg, 0.385 mmol) was added to the mixture and the mixture was stirred at 25° C. for 8 h. The mixture was quenched with water (20 mL) and dichloromethane (20 mL). * 3), and the organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (column: Welch Xtimate C18 150 * 25mm * The mixture was purified using a 500 mL / min column with a 5% elution buffer, mobile phase A: water (0.225% FA), mobile phase B: ACN, flow rate: 25 mL / min, gradient conditions 1% B to 30%). Pure fractions were collected and the solvent was evaporated under vacuum to give a residue. Pure fractions were collected and the solvent was evaporated under vacuum to give a residue. The residue was adjusted to pH=8 with a solution of 10% sodium hydroxide (20 mL) and diluted with dichloromethane (30 mL). * 3), the organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue, which was partitioned between acetonitrile (2 mL) and water (10 mL). The mixture was lyophilized to dryness to give the product (40 mg, 95.02% purity, 61.95% yield) as a colorless oil, which was mixed with fumaric acid (14.6 mg, 0.126 mmol), acetonitrile (12 mL) and water (4 mL) in a 50 mL round-bottom flask. The mixture was concentrated under reduced pressure to give a residue. The residue was partitioned between acetonitrile (3 mL) and water (10 mL). The solution was lyophilized to dryness to give compound 8 (27.02 mg, 99.99% purity, 49.48% yield) as a white solid.

[0389] Preparation of compound 9:

[0390] [ka]

[0391] HCl / dioxane (1 mL, 4 M) was added to a solution of intermediate 35 (150 mg, 0.210 mmol) in DCM (3 mL) and the mixture was stirred at 25° C. for 0.5 h. The solution was concentrated to give the crude product, which was partitioned between acetonitrile (3 mL) and water (10 mL). The solution was lyophilized to dryness to give compound 9 (99.54 mg, 98.85% purity, 65% yield) as a yellow solid.

[0392] The following compounds were synthesized by methods similar to those described above for compound 9.

[0393] [Table 10]

[0394] Preparation of compound 11:

[0395] [ka]

[0396] Intermediate 16 (100 mg, 0.190 mmol) was added to a solution of N-methylpropargylamine (26.2 mg, 0.380 mmol), MeOH (5 mL), and HOAc (34.2 mg, 0.570 mmol). The mixture was stirred at room temperature for 15 h. NaBH3CN (10.7 mg, 0.171 mmol) was then added to the solution and stirred for an additional 45 min. The mixture was chromatographed using Boston Prime C18 150 * 30mm * Purification by preparative HPLC using 5um (eluent: 55%-85% MeCN and water (0.05% NH3H2O+10mM NH4HCO3) afforded the pure product. The product was suspended in water (10mL) and frozen using dry ice / EtOH, then lyophilized to dryness to afford the title compound (18.52mg, 16% yield) as a white solid.

[0397] Preparation of compound 12:

[0398] [ka]

[0399] Intermediate 16 (100 mg, 0.190 mmol) was added to a solution of N-allylmethylamine (27 mg, 0.38 mmol), MeOH (1 mL), and HOAc (34.2 mg, 0.570 mmol). The mixture was then stirred at room temperature for 15 h. NaBH3CN (10.7 mg, 0.171 mmol) was then added to the solution and stirred for an additional 45 min. The mixture was then cooled to 100° C. using Boston Prime C18 150 * 30mm * Purification by preparative HPLC using 5um (eluent: 50%-80% MeCN and water (0.05% NH3H2O) gave the pure product. The product was suspended in water (10mL), frozen using dry ice / EtOH, then lyophilized to dryness to give the product (55.8mg, 51% yield) as a brown oil, which was mixed with fumaric acid (22.3mg, 0.192mmol) in MeCN (2mL) and HO (2mL). The mixture was then concentrated under reduced pressure to give a residue. The residue was partitioned between acetonitrile (1mL) and water (3mL). The solution was lyophilized to dryness to give compound 12 (28.19mg, 35% yield) as a white solid.

[0400] Preparation of compounds 13a and 13b:

[0401] [ka]

[0402] After adding a stir bar, intermediate 17 (190 mg, 0.351 mmol), 2-methoxy-N-methylethanamine (157 mg, 1.76 mmol), and anhydrous dichloromethane (10 mL) to a 40 mL glass bottle, the resulting mixture was stirred at 25° C. for 1 h, and then sodium triacetoxyborohydride (150 mg, 0.708 mmol) was added to the mixture. The reaction mixture was stirred at 25° C. for another 8 h. The mixture was quenched with a saturated solution of sodium bicarbonate (60 mL) and extracted with dichloromethane (30 mL×2). The combined organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (column: Boston Prime C18 150 * 30mm * Purification was performed using a 5 μm column, mobile phase A: water (0.05% NH3H2O), mobile phase B: acetonitrile, flow rate: 30 mL / min, gradient conditions 55% B to 85%). Pure fractions were collected and the solvent was evaporated under vacuum to give a residue. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The solution was lyophilized to dryness to give the product (150 mg, purity 97.102%, yield 67.5%) as a pale yellow oil.

[0403] Compound 13a (150 mg, 0.244 mmol) was separated by SFC (DAICEL CHIRAL TECHNOLOGIES (CHINA) CO., LTD.) (separation conditions: CHIRALPAK IG-3 (IG30CD-WE016) (0.46 cm ID x 15 cmL), mobile phase: ACN / DEA = 100 / 0.1 (V / V), 1.0 mL / min, column temperature: 35, nozzle pressure: 100 bar, nozzle temperature: 60, evaporator temperature: 20, trimmer temperature: 25, wavelength: UV 254 nm). The fraction of the first peak was collected and the solvent was evaporated under vacuum to obtain a residue. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The solution was lyophilized to dryness to give a pale yellow oil (60.0 mg, purity 84.967%, yield 34.0%), which was purified by preparative HPLC (column: Boston Prime C18 150 * 30mm *Further purification was performed with 5 um, mobile phase A: water (0.05% NH3H2O ​​+ 10 mM NH4HCO3), mobile phase B: acetonitrile, flow rate: 25 mL / min, gradient conditions 65%B to 95%). Pure fractions were collected and the solvent was evaporated under vacuum to give a residue. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The solution was lyophilized to dryness to give compound 1a (42.2 mg, purity 99.047%, yield 69.7%) as a yellow oil, which was mixed with fumaric acid (16.0 mg, 0.138 mmol) and acetonitrile (5 mL) in a 50 mL round bottom flask, and the resulting mixture was stirred at 25 °C for 1 h. The resulting mixture was concentrated under reduced pressure to give a residue. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The solution was lyophilized to dryness to give compound 13a (46.78 mg, 97.40% purity, 78.3% yield) as a white solid.

[0404] The fractions of the second peak were collected and the solvent was evaporated under vacuum to give a residue. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The solution was lyophilized to dryness to give the crude material (60.0 mg, 94.954% purity, 38.0% yield) as a pale yellow oil, which was purified by preparative HPLC (column: Boston Prime C18 150 * 30mm *The compound was purified by HPLC using a 500 sq. m. column with 5 um, mobile phase A: water (0.05% NH3H2O ​​+ 10 mM NH4HCO3), mobile phase B: acetonitrile, flow rate: 25 mL / min, gradient conditions 65%B to 95%). Pure fractions were collected and the solvent was evaporated under vacuum to give a residue. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The solution was lyophilized to dryness to give compound 1b (47.5 mg, 100% purity, 79.2% yield) as a yellow oil, which was mixed with fumaric acid (18.0 mg, 0.155 mmol) and acetonitrile (5 mL) in a 50 mL round bottom flask, and the resulting mixture was stirred at 25°C for 1 h. The resulting mixture was concentrated under reduced pressure to give a residue. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The solution was lyophilized to dryness to give compound 13b (43.36 mg, purity 97.35%, yield 64.5%) as a white powder.

[0405] Preparation of compound 14:

[0406] [ka]

[0407] To a solution of intermediate 38 (crude from previous step, 200 mg, 0.465 mmol) in MeOH (2 mL, 0.791 g / mL, 49.372 mmol), 6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-one (140.291 mg, 0.697 mmol) was added followed by sodium acetate (114.3 mg, 1.394 mmol) and stirred at room temperature for 10 min, then sodium cyanoborohydride (87.6 mg, 1.394 mmol) was added and stirred at room temperature for 16 h. The reaction mixture was then purified by HPLC (Preparation method: waters X-bridge C18 (5 μm 19 * 150 mm), mobile phase A: water (0.1% ammonium bicarbonate), mobile phase B: acetonitrile, UV: 214 nm, flow rate: 15 mL / min, gradient: 20-70% (% B)) to give compound 14 (48 mg, purity 98%, yield 16.4%).

[0408] Preparation of compounds 16, 17 and 18:

[0409] [ka]

[0410] A mixture of compound 14 (45 mg, 0.073 mmol) was purified by SFC (separation conditions: DAICEL CHIRALPAK IG (250 mm * 30mm, 10um), mobile phase: A: supercritical CO2, B: 0.1%NH 3. The mixture was separated by elution with HO IPA, A:B=55:45, 70 mL / min. Three fractions were obtained. The first fraction was collected and the solvent was evaporated under vacuum. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The solution was lyophilized to dryness to give a colorless oil (20 mg, 94% pure by LCMS), which was purified by SFC (separation conditions: DAICEL CHIRALPAK IG (250 mm * Further separation was performed using a 30 mm, 10 um mobile phase: A: supercritical CO2, B: 0.1% NH3H2O ​​EtOH A:B=75:25, 60 mL / min). The pure fractions of the first peak were collected and the solvent was evaporated under vacuum. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The solution was lyophilized to dryness to give compound 18 (3.51 mg, 87.38% purity, 15.34% yield) as a colorless oil.

[0411] The second fraction was collected and the solvent was evaporated under vacuum. The residue was suspended in water (10 mL) and the mixture was frozen using dry ice / ethanol, then lyophilized to dryness to give compound 16 (2.0 mg, purity 81.66%, yield 3.63%) as a colorless oil.

[0412] The third fraction was collected and the solvent was evaporated under vacuum.The residue was suspended in water (10 mL) and the mixture was frozen using dry ice / ethanol, then lyophilized to dryness to give compound 17 (2.79 mg, purity 79.48%, yield 4.93%) as a colorless oil.

[0413] Preparation of compound 19:

[0414] [ka]

[0415] A stir bar, intermediate 45 (270 mg, 0.42 mmol), wet palladium on activated carbon (100 mg, w / w%=10%, containing 50% water) and anhydrous methanol (20 mL) were added to a hydrogenation bottle, and then triethylamine (126 mg, 1.25 mmol) was added to the mixture. The suspension was degassed under vacuum, purged with N2 atmosphere three times, and then purged with hydrogen three times. The resulting mixture was stirred at 25° C. under hydrogen (15 psi) for 12 hours. The mixture was filtered through a pad of Celite® and washed with filter cake (20 mL×3). The combined filtrate was concentrated under reduced pressure to give the crude product, which was dissolved in dichloromethane (50 mL). The organic layer was washed with 10% aqueous NaOH (20 mL), water (20 mL) and brine (20 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the product (99.1 mg, 95.60% purity, 37.1% yield) as a yellow solid, which was mixed with fumaric acid (37.4 mg, 0.32 mmol) and MeCN (2 mL) in a 50 mL round-bottom flask. The reaction mixture was stirred at 25° C. for 30 min. The reaction mixture was concentrated under reduced pressure to give a residue, which was partitioned between acetonitrile (1 mL) and water (3 mL). The mixture was lyophilized to dryness to give compound 19 (79.37 mg, 88.30% purity, 51.4% yield) as a yellow solid.

[0416] Preparation of compound 20:

[0417] [ka]

[0418] Intermediate 48 (60 mg, 0.0967 mmol) and paraformaldehyde (2.902 mg, 0.0967 mmol) were added in MeOH (4 mL, 0.791 g / mL, 98.745 mmol). The mixture was stirred at room temperature for 0.5 h. NaBH3CN (6.074 mg, 0.0967 mmol) was added. The mixture was stirred for 16 h. The mixture was purified by flash column (C18, CH3CN:H2O 5:95 to 30:70, HCOOH as buffer) to give compound 20 (4 mg, 5.2% yield).

[0419] Preparation of compound 21:

[0420] [ka]

[0421] To a solution of intermediate 51 (200 mg, 0.416 mmol) and intermediate 41 (185 mg, 0.622 mmol) in dry acetonitrile (10 mL) was added DBU (190 mg, 1.25 mmol). The reaction mixture was stirred at 25° C. for 12 h. The reaction mixture was poured into water (50 mL) and extracted with dichloromethane (30 mL×3 times). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue, which was purified by preparative HPLC (column: Boston Green ODS 150 * 30mm * The mixture was purified using a 5 μm column, mobile phase A: water (0.225% FA), mobile phase B: acetonitrile, flow rate: 35 mL / min, gradient conditions 10% B to 40%). Pure fractions were collected and the solvent was evaporated under vacuum. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The mixture was lyophilized to dryness to give the desired compound (150 mg, purity 98.35%, yield 52.3%).

[0422] The product (89.2 mg, 0.131 mmol), fumaric acid (30.51 mg, 0.263 mmol) and MeCN (2 mL) were added to a 50 mL round bottom flask. The reaction mixture was stirred at 25° C. for 30 min. The reaction mixture was concentrated under reduced pressure to give a residue which was partitioned between acetonitrile (1 mL) and water (3 mL). The mixture was lyophilized to dryness to give compound 21 (78.99 mg, 98.86% purity, 65.2% yield) as a yellow solid.

[0423] Preparation of compound 22:

[0424] [ka]

[0425] Triethylamine (50.6 mg, 0.500 mmol) was added to a solution of intermediate 54a (70.0 mg, 0.100 mmol) in dry dichloromethane (5 mL). Then, aqueous formaldehyde solution (40.6 mg, 0.500 mmol) was added. The reaction mixture was stirred at 25° C. for 30 min, followed by the addition of sodium triacetoxyborohydride (42.4 mg, 0.200 mmol). The reaction mixture was stirred at 25° C. for another 12 h. The reaction mixture was diluted with dichloromethane (50 mL), a saturated solution of sodium bicarbonate (50 mL) was added, and the mixture was extracted with dichloromethane (30 mL×3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give a residue, which was purified by preparative HPLC (column: Boston Prime C18 150 * 25mm * The mixture was purified using a 5 μm column, mobile phase A: water (0.05% NH3H2O), mobile phase B: acetonitrile, flow rate: 35 mL / min, gradient conditions 51% B to 81%). Pure fractions were collected and the solvent was evaporated under vacuum. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The mixture was lyophilized to dryness to give compound 22 (30.79 mg, purity 98.83%, yield 50.7%) as a white powder.

[0426] Triethylamine (50.6 mg, 0.500 mmol) was added to a solution of intermediate 54b (70.0 mg, 0.100 mmol) in dry dichloromethane (5 mL). Then, aqueous formaldehyde (40.6 mg, 0.500 mmol) was added. The reaction mixture was stirred at 25° C. for 30 min, followed by the addition of sodium triacetoxyborohydride (42.4 mg, 0.200 mmol). The reaction mixture was stirred at 25° C. for another 12 h. The reaction mixture was diluted with dichloromethane (50 mL), a saturated solution of sodium bicarbonate (50 mL) was added, and the mixture was extracted with dichloromethane (30 mL×3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give a residue, which was purified by preparative HPLC (column: Boston Prime C18 150 * 30mm * The mixture was purified using a 5 μm column, mobile phase A: water (0.05% NH3H2O), mobile phase B: acetonitrile, flow rate: 35 mL / min, gradient conditions 50% B to 80%). Pure fractions were collected and the solvent was evaporated under vacuum. The residue was partitioned between acetonitrile (2 mL) and water (10 mL). The mixture was lyophilized to dryness to give compound 23 (29.02 mg, purity 99.54%, yield 48.1%) as a white powder.

[0427] The following compounds were synthesized by methods similar to those described above for compounds 22 and 23.

[0428] [Table 11]

[0429] LCMS (Liquid chromatography / Mass spectrometry) General Procedure High performance liquid chromatography (HPLC) measurements were performed using the LC pump, diode-array (DAD) or UV detector and column specified in each method. Additional detectors were included if necessary (see methods table below).

[0430] The flow from the column was delivered to a mass spectrometer (MS) configured with an atmospheric pressure ion source. It is within the knowledge of one skilled in the art to set tuning parameters (e.g., scan range, dwell time, etc.) to obtain ions that allow identification of the nominal monoisotopic molecular weight (MW) of the compound. Data collection was performed with appropriate software.

[0431] Compounds were identified based on their experimental retention times (R t ) and ions. Unless otherwise specified in the tables of data, the molecular ions reported are [M+H] + (protonated molecule) and / or [MH] - (deprotonated molecule). If the compound is not directly ionizable, the type of adduct is specified (i.e., [M+NH4] + , [M+HCOO] - , etc.). For molecules with multiple isotopic patterns (Br, Cl), the values ​​reported are those obtained for the lowest isotopic mass. All results were obtained with experimental uncertainties typically associated with the methods used.

[0432] In the following, "SQD" means single quadrupole detector, "RT" means room temperature, "BEH" means bridged ethylsiloxane / silica hybrid, "HSS" means high strength silica, and "DAD" means diode array detector.

[0433] [Table 12]

[0434] [Table 13]

[0435] Pharmacological part 1) Menin / MLL homogenous time-resolved fluorescence (HTRF) assay To a non-treated white 384-well microtiter plate, 40 nL of 200× test compound in DMSO and 4 μL of 2× terbium chelate-labeled menin (preparation see below) in assay buffer (40 mM Tris·HCl, pH 7.5, 50 mM NaCl, 1 mM DTT (dithiothreitol) and 0.05% Pluronic F-127) were added. After incubation of the test compound and terbium chelate-labeled menin for 30 min at ambient temperature, 4 μL of 2× FITC-MBM1 peptide (FITC-β-alanine-SARWRFPARPGT-NH2) (where "FITC" stands for fluorescein isothiocyanate) in assay buffer was added, the microtiter plate was centrifuged at 1000 rpm for 1 min, and the assay mixture was incubated at ambient temperature for 15 min. The relative amount of menin·FITC-MBM1 complex present in the assay mixture is determined by measuring the homogeneous time-resolved fluorescence (HTRF) of the terbium / FITC donor / acceptor fluorophore pair using an EnVision microplate reader (excitation 337 nm / terbium emission 490 nm / FITC emission 520 nm) at ambient temperature. The extent of fluorescence resonance energy transfer (HTRF value) is calculated as the ratio of the fluorescence emission intensities of the FITC and terbium fluorophores (F em 520nm / F em The binding assay is expressed as a 490 nm (490 nm) window. The final concentrations of reagents in the binding assay are 200 pM terbium chelate-labeled menin, 75 nM FITC-MBM1 peptide and 0.5% DMSO in assay buffer. Dose-response titrations of test compounds are typically performed using an 11-point 4-fold serial dilution scheme starting at 10 μM.

[0436] Compound potency was determined by first calculating the % inhibition at each compound concentration according to Equation 1. Inhibition% = ((HC-LC)-(HTRF 化合物 -LC)) / (HC-LC)) * 100 (formula 1) where LC and HC are the HTRF values ​​of the assay in the presence or absence of a saturating concentration of a compound that competes with FITC-MBM1 for binding to menin, and HTRF 化合物 is the HTRF value measured in the presence of test compound. HC and LC HTRF values ​​represent the average of at least 10 replicates per plate. For each test compound, the % inhibition values ​​were plotted against the logarithm of the test compound concentration, and these data were fitted to equation 2 to obtain the IC 50 We derived the value: Inhibition% = bottom + (top - bottom) / (1 + 10^((logIC 50 -log[cmpd]) * h))(Formula 2) where bottom and top are the lower and upper asymptote of the dose-response curve, respectively, and IC 50 is the concentration of compound that produces 50% inhibition of the signal, and h is the Hill coefficient. IC<0.1 nM in the HTRF assay 50 Values ​​are reported in the table below as 0.1 nM (limit of detection).

[0437] Preparation of terbium cryptate labeling of menin: Menin (aa 1-610-6xhis tag, 2.3 mg / mL in 20 mM Hepes (2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid), 80 mM NaCl, 5 mM DTT (dithiothreitol), pH 7.5) was labeled with terbium cryptate as follows: 200 μg of menin was buffer exchanged into 1x Hepes buffer. 6.67 μM of menin was incubated with an 8-fold molar excess of NHS (N-hydroxysuccinimide)-terbium cryptate for 40 min at room temperature. Half of the labeled protein was purified from the free label by running the reaction on a NAP5 column with elution buffer (0.1 M Hepes, pH 7 + 0.1% BSA (bovine serum albumin)). The remaining half was eluted with 0.1 M phosphate buffered saline (PBS), pH 7. 400 μl of each eluate was collected, aliquoted, and frozen at −80° C. The final concentrations of terbium-labeled menin protein were 115 μg / mL in Hepes buffer and 85 μg / mL in PBS buffer, respectively.

[0438] Menin protein sequence (SEQ ID NO:1) MGLKAAQKTLFPLRSIDDVVRLFAAELGREEPDLVLLSLVLGFVEHFLAVNRVIPTNVPELTFQPSPAPDPPGGLTYFPVADLSIIAALYARFTAQIRGAVDLSLYPREGGVSSRELVKKVSDVIWNSLSRSYFKDRAHIQSLFSFITGTKLDS SGVAFAVVGACQALGLRDVHLALSEDHAWVVFGPNGEQTAEVTWHGKGNEDRRGQTVNAGVAERSWLYLKGSYMRCDRKMEVAFMVCAINPSIDLHTDSLELLQLQQKLLWLLYDLGHLERYPMALGNLADLEELEPTPGRPDPLTLYHKGIAS AKTYYRDEHIYPYMYLAGYHCRNRNVREALQAWADTATVIQDYNYCREDEEIYKEFFEVANDVIPNLLKEAASLLEAGEERPGEQSQGTQSQGSALQDPECFAHLLRFYDGICKWEEGSPTPVLHVGWATFLVQSLGRFEGQVRQKVRIVSREA EAAEAEEPWGEEAREGRRRGPRRESKPEEPPPPKKPALDKGLGTGQGAVSGPPRKPPGTVAGTARGPEGGSTAQVPAPAASPPPEGPVLTFQSEKMKGMKELLVATKINSSAIKLQLTAQSQVQMKKQKVSTPSDYTLSFLKRQRKGLHHHHHH

[0439] 2) Proliferation assay The anti-proliferative effect of menin / MLL protein / protein interaction inhibitor test compound was evaluated in human leukemia cell line. Cell line MOLM14 has MLL translocation and expresses MLL fusion protein MLL-AF9, as well as wild-type protein from the second allele. MLL-rearranged cell line (e.g., MOLM14) shows stem cell-like HOXA / MEIS1 gene expression signature. To exclude compounds that show general cytotoxic effects, KO-52 was used as a control cell line that contains two MLL (KMT2A) wild-type alleles.

[0440] MOLM14 cells were cultured in RPMI-1640 (Sigma Aldrich) supplemented with 10% heat-inactivated fetal bovine serum (HyClone), 2 mM L-glutamine (Sigma Aldrich), and 50 μg / ml gentamicin (Gibco). The KO-52 cell line was grown in α-MEM (Sigma Aldrich) supplemented with 20% heat-inactivated fetal bovine serum (HyClone), 2 mM L-glutamine (Sigma Aldrich), and 50 μg / ml gentamicin (Gibco). During culture, cells were maintained at 300,000 to 2.5 million cells / ml and did not exceed 20 passages.

[0441] To evaluate antiproliferative effects, 200 MOLM14 cells or 300 KO-52 cells were seeded in 200 μl of medium per well of a 96-well round-bottom ultra-low attachment plate (Costar, Cat. No. 7007). The cell seeding number was selected based on the growth curve to ensure linear growth throughout the experiment. Test compounds were added at different concentrations and DMSO content was normalized to 0.3%. Cells were incubated at 37°C and 5% CO2 for 8 days. Spheroid-like growth was measured in real time by live cell imaging (IncuCyteZOOM, Essenbio, 4x objective) with image acquisition on day 8. Confluence (%) as a measure of spheroid size was determined using the integrated analysis tool.

[0442] To determine the effect of test compounds over time, the confluence in each well was calculated as a measure of spheroid size. The confluence of the highest dose of reference compound was used as the baseline for LC (Low control), and the confluence of DMSO-treated cells was used as 0% cytotoxicity (High Control, HC).

[0443] Absolute IC 50 Values ​​were calculated as percentage change in confluence as follows: LC = low control: cells treated with, for example, 1 μM of the cytotoxic agent staurosporine, or cells treated with, for example, a high concentration of a surrogate reference compound HC = High control: average confluence (%) (DMSO-treated cells) %Effect=100-(100 * (Sample-LC) / (HC-LC)) IC was calculated using GraphPad Prism (version 7.00). 50 For plots of % effect versus Log10 compound concentration, a dose-response equation was used with variable slope, max fixed at 100% and min fixed at 0%.

[0444] [Table 14]

Claims

1. Formula (I): 【Chemical 1】 [wherein, R １ａ is -C(=O)-NR ｘａ R ｘｂ , Het, or 【Chemical Formula 2】 represents, Het represents a 5- or 6-membered monocyclic aromatic ring containing 1, 2 or 3 nitrogen atoms and optionally a carbonyl moiety, The 5- or 6-membered monocyclic aromatic ring is optionally substituted with 1, 2 or 3 substituents selected from the group consisting of C ３～６ cycloalkyl and C １～４ alkyl R ｘａ and R ｘｂ are each independently hydrogen; １～４ Alkyl; C ３～６ Cycloalkyl; C substituted with 1, 2 or 3 halo atoms １～４ Alkyl; and one -OH, -OC １～４ Alkyl or NR １１ｃ R １１ｄ C substituted with １～４ is selected from the group consisting of alkyl, R １ｂ represents F or Cl, R １ｃ represents H or a halo, Y １ is -CR ５ａ R ５ｂ -, -O- or -NR ５ｃ -, and represents R ２ is selected from the group consisting of hydrogen, halo, C １～４ alkyl, -O-C １～４ alkyl, and -NR ７ａ R ７ｂ and is selected from the group consisting of U represents N or CH, n1, n2, n3 and n4 are each independently selected from 1 and 2, X １ represents CH, and X ２ represents N, R ４ is C １～５ alkyl, [Chemical Formula 3] represents, R ５ａ 、 R ５ｂ 、 R ５ｃ 、 R ７ａ 、 and R ７ｂ are each independently selected from the group consisting of hydrogen, C １～４ alkyl, and C ３～６ cycloalkyl, R ３ is -C １～６ alkyl-NR ８ａ R ８ｂ , -C １～６ alkyl-C(=O)-NR ９ａ R ９ｂ , -C １～６ alkyl-OH, or, -C １～６ alkyl-NR １１ -C(=O)-O-C １～４ alkyl-O-C(=O)-C １～４ represents alkyl, R ３ C in the definition of １～４ alkyl or C １～６ each of the alkyl moieties is independently substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH, and -O-C １～４ alkyl and may be substituted R ８ａ and R ８ｂ are each independently hydrogen; C １～６ alkyl; -C(=O)-C １～４ alkyl; -C(=O)-O-C １～４ alkyl; -C(=O)-NR １２ａ R １２ｂ ; and -OH, cyano, halo, -C≡C, -CH=CH, -S-C １～４ alkyl, -S(=O) ２ -C １～４ alkyl, -S(=O) ２ -NR １１ａ R １１ｂ ; -O-C １～４ alkyl, -C(=O)-NR １０ａ R １０ｂ ; -NR １０ｃ -C(=O)-C １～４ alkyl, one NR １１ａ R １１ｂ substituted -O-C １～４ alkyl, and -O-C １～４ alkyl substituted with 1, 2 or 3 halo atoms, each independently selected from the group consisting of 1, 2 or 3 substituents; C １～６ alkyl; selected from the group consisting of, R ９ａ 、 R ９ｂ 、 R １０ａ 、 R １０ｂ 、 R １０ｃ 、 R １１ 、 R １１ａ 、 R １１ｂ 、 R １２ａ 、 and R １２ｂ are each independently selected from the group consisting of hydrogen and C １～６ alkyl, R １１ｃ and R １１ｄ are each independently selected from the group consisting of hydrogen, C １～６ alkyl and -C(=O)-C １～４ alkyl] a compound of, or a tautomer or stereoisomeric form thereof, or a pharmaceutically acceptable salt or solvate thereof, provided that: a) R １ａ represents Het, and the 5- or 6-membered monocyclic aromatic ring is substituted with three substituents selected from the group consisting of C ３～６ cycloalkyl and C １～４ alkyl; b) R １ａ is -C(=O)-NR ｘａ R ｘｂ wherein R ｘａ is selected from the group consisting of one -OH, -OC １～４ alkyl or NR １１ｃ R １１ｄ substituted C １～４ alkyl, and C １～４ alkyl substituted with one, two or three halo atoms c) R １ｃ represents a halo, d) R ４ wherein R is other than isopropyl, e) R ３ is -C １～６ alkyl-NR ８ａ R ８ｂ where R ８ａ is -C≡C, -CH=CH, -S-C １～４ alkyl, -S(=O) ２ -NR １１ａ R １１ｂ , one NR １１ａ R １１ｂ substituted -O-C １～４ alkyl, and -O-C １～４ alkyl substituted with one, two or three substituents each independently selected from the group consisting of one, two or three substituents selected from -O-C １～６ alkyl substituted with one, two or three halo atoms is at least one of the following conditions: is satisfied, a compound of formula (I), or a tautomer or stereoisomeric form thereof, or a pharmaceutically acceptable salt or solvate thereof.

2. R １ａ is -C(=O)-NR ｘａ R ｘｂ and represents R ｘａ and R ｘｂ are each independently C １～４ alkyl; and C substituted with one —OH or NR １１ｃ R １１ｄ alkyl; and are selected from the group consisting of: １～４ ​ R １ｂ represents F, and Y １ represents -O-, R ２ represents hydrogen, and R ４ represents C １～５ alkyl, and R ３ is -C １～６ alkyl -NR ８ａ R ８ｂ represents, R ３ C in the definition of １～６ the alkyl moiety may be substituted with 1, 2 or 3 -OH substituents, R ８ａ and R ８ｂ are each independently hydrogen; C １～６ alkyl; and -C≡C, -CH=CH, -S-C １～４ alkyl, -S(=O) ２ -NR １１ａ R １１ｂ -O-C １～４ alkyl, one NR １１ａ R １１ｂ substituted -O-C １～４ alkyl, and -O-C １～４ alkyl substituted with 1, 2 or 3 halo atoms; C １～６ alkyl; selected from the group consisting of, R １１ａ and R １１ｂ represent hydrogen, and R １１ｃ and R １１ｄ are each independently selected from the group consisting of hydrogen and -C(=O)-C １～４ alkyl, provided that: a) R １ａ is -C(=O)-NR ｘａ R ｘｂ represents, and R ｘａ is selected from the group consisting of C １１ｃ alkyl substituted with one -OH or NR １１ｄ R １～４ and b) R １ｃ represents a halo, c) R ４ is tert-butyl, d) R ３ is -C １～６ alkyl-NR ８ａ R ８ｂ wherein R ８ａ is -C≡C, -CH=CH, -S-C １～４ alkyl, -S(=O) ２ -NR １１ａ R １１ｂ , one NR １１ａ R １１ｂ substituted -O-C １～４ alkyl, and -O-C １～４ alkyl substituted with one, two or three substituents each independently selected from the group consisting of alkyl and -O-C １～６ alkyl substituted with one, two or three substituents each independently selected from the group consisting of alkyl and -O-C at least one of the following conditions: is satisfied, the compound according to claim 1. R １ａ wherein R is -C(=O)-NR ｘａ R ｘｂ The compound according to claim 1.

3. R １ａ is -C(=O)-NR ｘａ R ｘｂ and represents Y １ represents -O-, and U represents N, n1 is 1, n2 is 2, n3 is 1, and n4 is 1. R １ｂ represents F, and R １ｃ represents H, and R ２ represents hydrogen, and R ４ represents C １～５ alkyl, and R ３ is -C １～６ alkyl - NR ８ａ R ８ｂ The compound according to claim 1, wherein

5. The compound according to claim 1, wherein U represents N.

6. Y １ The compound according to claim 1, wherein Y represents -O-.

7. R １ｂ The compound according to claim 1, wherein R represents F.

8. A pharmaceutical composition comprising the compound according to any one of claims 1 to 7 and a pharmaceutically acceptable carrier or diluent.

9. A process for preparing the pharmaceutical composition according to claim 8, comprising mixing a pharmaceutically acceptable carrier with a therapeutically effective amount of the compound according to any one of claims 1 to 7.

10. A pharmaceutical composition comprising the compound according to any one of claims 1 to 7 for use as a medicament.

11. A pharmaceutical composition comprising the compound according to any one of claims 1 to 7 for use in the prevention or treatment of cancer, myelodysplastic syndrome (MDS) and diabetes.

12. The pharmaceutical composition according to claim 11, wherein the cancer is selected from leukemia, myeloma or solid tumor cancer, such as prostate cancer, lung cancer, breast cancer, pancreatic cancer, colon cancer, liver cancer, melanoma and glioblastoma.

13. The pharmaceutical composition according to claim 12, wherein the leukemia is selected from acute leukemia, chronic leukemia, myeloblastic leukemia, myeloid leukemia, lymphoblastic leukemia, lymphocytic leukemia, acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), T-cell prolymphocytic leukemia (T-PLL), large granular lymphocytic leukemia, hairy cell leukemia (HCL), MLL-rearranged leukemia, MLL-PTD leukemia, MLL amplified leukemia, MLL positive leukemia, and leukemia showing HOX / MEIS1 gene expression signature.

14. The pharmaceutical composition according to claim 11 for use in the prevention or treatment of cancer.