EA045364B1 - SPIROBICYCLIC INHIBITORS OF MENIN-MLL INTERACTION - Google Patents

Description

The present invention relates to pharmaceutical agents useful for therapy and/or prophylaxis in a mammal, and in particular to spirobicyclic compounds, a pharmaceutical composition containing such compounds, and their use as inhibitors of menin-MLL protein-protein interactions useful for the treatment of diseases , such as cancer, myelodysplastic syndrome (MDS) and diabetes.

BACKGROUND OF THE INVENTION

Chromosomal rearrangements affecting the mixed lineage leukemia associated gene (MLL; MLL1; KMT2A) result in aggressive acute leukemias across all age groups and are still virtually incurable diseases, highlighting the urgent need for new therapeutic approaches . Types of acute leukemia involving such MLL chromosomal translocations are lymphoid, myeloid or biphenotypic disease and account for 5 to 10% of acute leukemias in adults and approximately 70% in children (Marschalek, Br J Haematol 2011. 152(2), 141-54; Tomizawa et al., Pediatr Blood Cancer 2007. 49(2), 127-32).

MLL is a histone methyltransferase that methylates histone H3 at lysine 4 (H3K4) and functions in multiprotein complexes. The use of inducible loss-of-function alleles of Mll1 has demonstrated that Mll1 plays an important role in hematopoietic stem cell (HSC) maintenance and B cell development, although its histone methyltransferase activity is not essential for hematopoiesis (Mishra et al., Cell Rep 2011. 7 (4), 1239-47).

To date, MLL fusions with over 60 different partners have been reported and have been associated with leukemia formation/progression (Meyer et al., Leukemia 2013. 27, 2165-2176). Interestingly, the SET domain (Su(var)3-9, zeste and trithorax enhancer) of MLL is not retained in the chimeric proteins but is replaced by a fusion partner (Thiel et al., Bioessays 2012. 34, 771-80). Recruitment of chromatin-modifying enzymes such as Dot1L and/or the pTEFb complex by a fusion partner results in enhanced transcription and transcriptional elongation of MLL target genes, including the HOXA genes (e.g., HOXA9) and the HOX cofactor MEIS1 as the most significant. Aberrant expression of these genes, in turn, blocks the differentiation of hematopoietic cells and enhances proliferation.

Menin, which is encoded by the multiple endocrine neoplasia type 1 associated gene (MEN1), is expressed ubiquitously and is predominantly localized to the nucleus. It has been shown to interact with numerous proteins and is therefore involved in a variety of cellular processes. The most studied function of menin is its role as an oncogenic cofactor of MLL-based fusion proteins. Menin interacts with two motifs in the N-terminal fragment of MLL, which is conserved in all fusion proteins, MBM1 (menin binding motif 1) and MBM2 (Thiel et al., Bioessays 2012. 34, 771-80). The menin-MLL interaction results in the formation of a new interaction surface for lens epithelial growth factor (LEDGF). Although MLL binds directly to LEDGF, the presence of menin is required for stable interaction between MLL and LEDGF and gene-specific recruitment of the MLL complex to chromatin via the PWWP domain of LEDGF (Cermakova et al., Cancer Res 2014. 15, 5139-51; Yokoyama & Cleary , Cancer Cell 2008. 8, 36-46). In addition, numerous genetic studies have shown that menin is clearly required for oncogenic transformation by MLL-based fusion proteins, implicating the menin-MLL interaction as an attractive therapeutic target. For example, conditional deletion of Men1 prevents leucomogenesis in bone marrow progenitor cells ectopically expressing MLL fusion products (Chen et al., Proc Natl Acad Sci 2006. 103, 1018-23). Likewise, genetic disruption of the menin-MLL fusion product interaction through loss-of-function mutations suppresses the oncogenic properties of MLL fusion proteins, blocks leukemia development in vivo, and reverses the differentiation block of MLL-transformed leukemic blast cells. These studies also showed that menin is required to maintain HOX gene expression by MLL-based fusion proteins (Yokoyama et al., Cell 2005. 123, 207-18). In addition, small molecule inhibitors of the menin-MLL interaction have been developed, demonstrating that this protein-protein interaction is useful as a drug target, and have also been demonstrated to be effective 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 of MLL1 during normal hematopoiesis (Li et al., Blood 2013. 122, 2039-2046), such data establish disruption of the menin-MLL interaction as a promising new therapeutic approach for the treatment of leukemia with MLL rearrangement and other cancers with an active HOX/MEIS1 gene expression profile. For example, internal partial tandem duplication (PTD) in the 5′ region of the MLL gene is another major aberration that is found predominantly de novo and in secondary AML and myelodysplastic syndromes. Although the molecular mechanism and biological function of MLL-PTD are not well understood, new therapeutic strategies targeting the menin interaction

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MLL may also be effective in treating MLL-PTD-associated leukemias. In addition, castration-resistant prostate cancer has been shown to be dependent on the menin-MLL interaction (Malik et al., Nat Med 2015. 21, 344-52).

Several literature sources describe inhibitors that specifically target the menin-MLL interaction: WO2011029054, J Med Chem 2016, 59, 892-913 thienopyrimidine and benzodiazepine; WO2014164543 describes thienopyrimidine and thienapyridine derivatives; in Nature Chemical Biology March 2012, 8, 277-284 and Ren, J.; et al. Bioorg Med Chem Lett (2016), http://dx.doi.Org/10.1016/j.bmcl.2016.07.074 thienopyrimidine derivatives are described; J Med Chem 2014, 57, 1543-1556 describes hydroxy- and aminomethylpiperidine derivatives; and Future Med Chem 2014, 6, 447-462 discusses small molecule and peptidomimetic compounds. WO2017112768 describes inhibitors of the menin-MLL interaction.

Description of the invention

The present invention relates to new compounds of formula (I):

and to their tautomers and stereoisomeric forms, where

R ¹ represents CF3;

R ² is selected from the group consisting of hydrogen and CH3;

L ¹ is a 7-10-membered saturated spiroheterobicyclic system containing one or two N atoms, provided that it is linked at N to a thienopyrimidinyl heterocycle;

And

-L ² -R ³ is selected from (a), (b), (c), (d), (f) or (g), where (a) L ² is selected from the group consisting of >CR ^4a R ^4b and -CHR ^4a CHR ⁵ -; Where

L ² is connected to the nitrogen atom L ¹ ; R ^4a is selected from the group consisting of hydrogen; -C(=O)NR ^7a R ^7b ;

C _1-4 alkyl, optionally substituted with a substituent selected from the group consisting of -OR ⁸ and -NR ^9a R ^9b ; and a C-linked 4-7 membered non-aromatic heterocyclyl containing at least one nitrogen atom;

R ⁵ is selected from the group consisting of hydrogen; -OR ⁶ and C _1-4 alkyl;

R ^4b is selected from the group consisting of hydrogen and methyl; or R ^4a and R ^4b together with the carbon atom to which they are attached form a C _3-5 cycloalkyl or a C 4-6 membered heterocyclyl containing an oxygen atom; wherein each of R ⁶ , R ^7a , R ^7b , R ⁸ , R ^9a and R ^9b is independently selected from the group consisting of hydrogen and C _2-4 alkyl substituted with a substituent selected from the group consisting of -OR ¹¹ and -NR ^10a R ^10b ; wherein each of R ^10a , R ^10b and R ¹¹ is independently selected from the group consisting of hydrogen and C _1-4 alkyl; And

R ³ is selected from the group consisting of Ar, Het ¹ , Het ² and a 7-10 membered saturated spirocarbobicyclic system; or (b) L ² represents >RC ^4c R ^4d where R ^4c and R ^4d represent hydrogen; and R^

R ³ represents ^R ' where R ^12a , R ^12b and R ^12c represents C _1-6alkyl ; or (c) -L2-R ³ is C _1-6 alkyl, optionally substituted with one, two or three fluorine substituents; or (d) L ² represents O and R ³ represents -CH ₂ -Ar; or (f) --L2-R ³ represents , where

R ¹⁸ is selected from the group consisting of hydrogen and C _1-4 alkyl; R ^18a is selected from the group consisting of hydrogen and fluorine;

R ^18b is selected from the group consisting of fluorine, -OC _1-4 alkyl and C _1-4 alkyl, optionally substituted with 1, 2 or 3 substituents representing a fluorine atom; or

R ^18a and R ^18b are bonded to the same carbon atom and together form a C _3-5 cycloalkyl; or

(g) --L2-R ³ represents

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Ar represents phenyl, which may be optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -OR ²⁴ and C ₁ . ₄ alkyl, optionally substituted -OR ²⁶ ;

Het ¹ is a monocyclic heteroaryl selected from the group consisting of pyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl, 4- or 5-thiazolyl, isothiazolyl and isoxazolyl; or bicyclic heteroaryl selected from the group consisting of indolyl, imidazopyridinyl; each of which may be optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -CN, -OR ²⁴ and C ₁ . ₄ alkyl, optionally substituted with a substituent selected from the group consisting of -CN, -OR ²⁶ and -NR ^27a R ^27b ; And

Het ² is a non-aromatic heterocyclyl, optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -CN and C1.4alkyl, optionally substituted -OR ²⁶ ; wherein each of R ²⁴ , R ²⁶ , R ^27a and R ^27b is independently selected from the group consisting of hydrogen; C1. ₄ alkyl and C ₂ . ₄ alkyl substituted -NR ^28a R ^28b ; Where

R ^28a and R ^28b represent hydrogen;

and pharmaceutically acceptable salts thereof;

provided that the following compounds and their pharmaceutically acceptable addition salts are excluded:

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It should be understood that the above condition applies to all embodiments of the present invention described later herein.

The present invention also relates to a pharmaceutical composition containing a therapeutically effective amount of a compound of formula (I), a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier or excipient.

Furthermore, the present invention relates to a compound of formula (I), a pharmaceutically acceptable salt or solvate thereof, for use as a drug, and a compound of formula (I), a pharmaceutically acceptable salt or solvate thereof, for use in the treatment or prevention of cancer, myelodysplastic syndrome ( MDS) and diabetes.

In a specific embodiment, the present invention relates to a compound of formula (I), a pharmaceutically acceptable salt or solvate thereof, for use in the treatment or prevention of cancer.

In a specific embodiment, said cancer is selected from leukemia, myeloma, or solid tumor cancer (e.g., prostate cancer, lung cancer, breast cancer, pancreatic cancer, colon cancer, liver cancer, melanoma and glioblastoma, etc.). d.). In some embodiments, the leukemia types include acute leukemia types, chronic leukemia types, myeloid leukemia types, myelogenous leukemia types, lymphoblastic leukemia types, lymphocytic leukemia types, acute myelogenous leukemia (AML) types, chronic myelogenous leukemia (CML) types, acute lymphoblastic leukemia types leukemia (ALL), types of chronic lymphocytic leukemia (CLL), types of T-cell prolymphocytic leukemia (T-PLL), leukemia of large granular lymphocytes, hairy cell leukemia (HCL), types of leukemia with MLL rearrangement, types of leukemia associated with MLL- PTD, types of leukemia with MLL amplification, types of MLL-positive leukemia, types of leukemia characterized by HOX/MEIS1 gene expression profiles, etc.

The present invention also relates to the use of a compound of formula (I), a pharmaceutically acceptable salt or solvate thereof, in combination with an additional pharmaceutical agent for use in the treatment or prevention of cancer, myelodysplastic syndrome (MDS) and diabetes.

Additionally, the present invention relates to a method for preparing a pharmaceutical composition in accordance with the present invention, which is characterized in that a pharmaceutically acceptable carrier is thoroughly mixed with a therapeutically effective amount of a compound of formula (I), a pharmaceutically acceptable salt or solvate thereof.

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The present invention also provides a product containing a compound of formula (I), a pharmaceutically acceptable salt or solvate thereof and an additional pharmaceutical agent, in the form of a combined preparation for simultaneous, separate or sequential use in the treatment or prevention of cancer, myelodysplastic syndrome (MDS) and diabetes.

In addition, the present invention provides a method of treating or preventing a cell proliferation disease in a warm-blooded animal, which comprises administering to said animal an effective amount of a compound of formula (I), a pharmaceutically acceptable salt or solvate thereof as defined herein, or a pharmaceutical composition or combinations defined herein.

Detailed Description of the Invention

The terms halogen or halogen as used herein mean fluorine, chlorine, bromine and iodine.

The prefix C _xy (where x and y are integers) as used herein refers to the number of carbon atoms in a given group. Thus, a C _1-6 alkyl group contains from 1 to 6 carbon atoms, a C _3-6 cycloalkyl group contains from 3 to 6 carbon atoms, and the like. The term C1 ₄ alkyl, as used herein as a group or part of a group, means a straight or branched chain saturated hydrocarbon radical containing from 1 to 4 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, etc.

The term _C2-4 alkyl, as used herein as a group or part of a group, means a straight or branched chain saturated hydrocarbon radical containing from 2 to 4 carbon atoms, such as ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, etc.

The term C _1-6 alkyl, as used herein as a group or part of a group, means a straight or branched chain saturated hydrocarbon radical containing from 1 to 6 carbon atoms, such as those defined for C _1-4 alkyl, and n -pentyl, n-hexyl, 2-methylbutyl, etc.

The term C _3-6 alkyl, as used herein as a group or part of a group, means a straight or branched chain saturated hydrocarbon radical containing from 3 to 6 carbon atoms, such as n-propyl, isopropyl, n-butyl, sec- butyl, tert-butyl, n-pentyl, n-hexyl, 2methylbutyl, etc.

The term C _3-5 cycloalkyl, as used herein as a group or part of a group, means a saturated cyclic hydrocarbon radical containing from 3 to 5 carbon atoms, such as cyclopropyl, cyclobutyl and cyclopentyl.

The term spirobicyclic, as used herein as a group or part of a group, means cyclic systems where two rings are connected at one atom. Examples of these systems are 7-10 membered saturated spiroheterobicyclic systems containing one or two N atoms, where one of the nitrogen atoms is always associated with the thienopyrimidinyl heterocycle in the compounds of formula (I) defined herein. Such spirocyclic systems include, but are not limited to, those formed by combining, for example, piperidine, pyrrolidine, azetidine and cyclobutane rings. Examples of such systems include, without limitation, (a), (b), (c), (d), (e), (f) and (g) below, and the like.

where a denotes the binding position with the thienopyrimidinyl heterocycle. One skilled in the art will appreciate that in these specific examples, the variants --L ² -R ³ defined herein when L ² is bonded to the nitrogen atom of L ¹ apply to examples (a)-(f); wherein the --L ² -R ³ variants defined herein, when L ² is bonded to the carbon atom L ¹ , apply to Example ⁽ g).

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Examples of 7-10 membered saturated spirocarbobicyclic systems include, but are not limited to

and so on.

In general, in all applications in the present invention, the term substituted is intended to mean, unless otherwise stated or clearly evident from the context, one or more hydrogen atoms, particularly 1 to 4 hydrogen atoms, more particularly 1 to 3 hydrogen atoms, preferably 1 or 2 hydrogen atoms, more preferably 1 hydrogen atom, on the atom or radical designated by the expression using substituted, are replaced by one selected from the group indicated, provided that the normal valence is not exceeded and that the substitution results in a chemically stable connection, i.e. a compound that is stable enough to withstand isolation to a suitable degree of purity from the reaction mixture.

Combinations of substituents and/or variables are permissible only if such combinations result in chemically stable compounds. A stable compound is meant to mean a compound that is sufficiently stable to withstand isolation to a suitable degree of purity from the reaction mixture.

One skilled in the art will understand that when an atom or radical is substituted with a substituent, it is understood that said atom or radical is substituted with one substituent selected from said group.

One skilled in the art will appreciate that the term optionally substituted means that the atom or radical designated using the expression optionally substituted may or may not be substituted (meaning substituted or unsubstituted, respectively).

If two or more substituents are present on a fragment, they may, if possible and unless otherwise indicated or clearly evident from the context, replace hydrogen atoms at the same atom, or they may replace hydrogen atoms at different atoms in the fragment.

One skilled in the art will appreciate that, unless otherwise stated or clearly apparent from the context, a substituent on a heterocyclyl group can replace any hydrogen atom on a ring carbon atom or on a ring heteroatom.

In the context of the present invention, saturated means fully saturated unless otherwise indicated.

The non-aromatic group includes unsaturated ring systems of non-aromatic nature, partially saturated and fully saturated carbocyclic and heterocyclic ring systems. The term partially saturated refers to rings where the ring structure(s) contains at least one multiple bond, for example a C=C, N=C bond. The term fully saturated refers to rings where there are no multiple bonds between the ring atoms. Thus, a non-aromatic heterocyclyl is a non-aromatic monocyclic or bicyclic system, unless otherwise specified, containing, for example, 3-12 ring members, more typically 5-10 ring members. Examples of monocyclic groups are those containing 4-7 ring members, more typically 5 or 6 ring members. Examples of bicyclic groups are those containing 8-12, more typically 9 or 10 ring members.

Non-limiting examples of monocyclic heterocyclyl systems containing at least one heteroatom selected from nitrogen, oxygen or sulfur (N, O, S) include, but are not limited to, 4-7 membered heterocyclyl systems such as azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl , piperazinyl, pyranyl, dihydropyranyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl and tetrahydro-2H-thiopyranyl-1,1-dioxide, in particular azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, pyranyl, dihydropyranyl, tetrahydropyranyl, morpho linyl and thiomorpholinyl. Non-limiting examples of bicyclic heterocyclyl systems containing at least one heteroatom selected from nitrogen, oxygen or sulfur (N, O, S),

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include, but are not limited to, octahydro-1H-indolyl, indolinyl, ' Unless otherwise stated, each may be linked to the remainder of the molecule of formula (I) via any available ring carbon atom (C-linked) or nitrogen atom (N-linked), and optionally they may be substituted, if possible, at carbon and/or nitrogen atoms in accordance with embodiments.

Examples of a C-linked 4-7 membered non-aromatic heterocyclyl containing at least one nitrogen atom include, but are not limited to, azetidinyl, pyrrolidinyl and piperidinyl linked to the remainder of the molecule via an accessible carbon atom.

The term C-linked 4-6 membered heterocyclyl containing an oxygen atom, as used herein alone or as part of another group, means a saturated cyclic hydrocarbon radical containing an oxygen atom that contains from 4 to 6 ring members, such as oxetanyl, tetrahydrofuranyl and tetrahydropyranyl.

In all cases where substituents are represented by a chemical structure, - means a bond for attachment to the remainder of the molecule of formula (I).

Lines (such as -) drawn to ring systems indicate that the bond can be attached to any of the suitable ring atoms.

Het ¹ and Het ² may be attached to the remainder of the molecule of formula (I) via any available ring carbon or nitrogen atom, as appropriate, unless otherwise indicated.

It will be appreciated that a saturated cyclic moiety may, if possible, contain substituents on both the carbon and N atoms unless otherwise indicated or clearly implied by the context.

It will be obvious that if L ² is >SO ₂ then this is equivalent to L ² being -SO2-. It will be obvious that if L ² is >CR ^4a R ^4b then this is equivalent to ^r4 \ s

L represents ·

For example, in compound 1, L ² is >CR ^4a R ^4b where both R ^4a and R ^4b are hydrogen.

Likewise, it will be obvious that if L ² represents >CR ^4c R ^4d , then this is equivalent to R ⁴ ^ _z R ^4d that L represents .

If any variable occurs more than once in any component, then each is defined independently.

If any variable appears more than once in any formula (for example, formula (I)), then each is defined independently.

The term subject as used herein refers to an animal, preferably a mammal (eg, a cat, dog, primate or human), more preferably a human, that is or has been the subject of treatment, observation or experiment.

The term therapeutically effective amount as used herein means that amount of an active compound or pharmaceutical agent that produces a biological or medical response in a tissue system in an animal or human that a researcher, veterinarian, physician, or other clinician seeks to produce, which includes relief or relief symptoms of the disease or disorder being treated.

The term composition is intended to cover a product containing certain ingredients in certain amounts, as well as any product that is obtained, directly or indirectly, from combinations of certain ingredients in certain amounts.

The term treatment as used herein is intended to include all modalities that may involve slowing, disrupting, suppressing or stopping the progression of a disease, but does not necessarily mean complete elimination of all symptoms.

The terms compound(s) of the present invention or compound(s) of the invention as used herein are intended to include the compounds of formula (I) and their pharmaceutically acceptable salts and solvates.

Any chemical formula used herein in which the bonds are shown only as solid lines rather than as solid wedges or dotted wedges, or otherwise shown as having a specific configuration (e.g., R, S) near one or more atoms , provides each possible stereoisomer or a mixture of two or more stereoisomers.

As used herein and below, it is intended that the term compound(s) of the formula

- 10 045364 (I) includes its tautomers and its stereoisomeric forms.

The terms stereoisomers, stereoisomeric forms, or stereochemically isomeric forms above or below are used interchangeably herein.

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.

Enantiomers are stereoisomers that are non-matching mirror images of each other when superimposed. A 1:1 mixture of enantiomer pairs is a racemate or racemic mixture.

Atropisomers (or atropisomers) are stereoisomers that have a specific spatial configuration formed as a result of limited rotation around a single bond due to significant steric hindrance. All atropisomeric forms of the compounds of formula (I) are intended to be included within the scope of the present invention.

Diastereomers (or diastereoisomers) are stereoisomers that are not enantiomers, i.e. they are not related as mirror images. If the compound contains a double bond, the substituents may be in the E- or Z-configuration.

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

Thus, the present invention includes enantiomers, atropisomers, diastereomers, racemates, E-isomers, Z-isomers, cis-isomers, trans-isomers and mixtures thereof, where chemically possible.

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

The absolute configuration is determined in accordance with the Cahn-Ingold-Prelog system. The configuration at an asymmetric atom is defined as either R or S. Isolated stereoisomers, the absolute configuration of which is unknown, can be designated as (+) or (-) depending on the direction in which they rotate the plane of polarization of light. For example, isolated enantiomers whose absolute configuration is unknown may be designated as (+) or (-) depending on the direction in which they rotate the plane of polarization of light.

If a particular stereoisomer is identified, this means that the specified stereoisomer contains essentially no other stereoisomers, i.e. is associated with less than 50%, preferably less than 20%, more preferably less than 10%, even more preferably less than 5%, in particular less than 2% and most preferably less than 1% of other stereoisomers. Thus, if a compound of formula (I), for example, is indicated as (R), this means that the compound contains substantially no (S)-isomer; if a compound of formula (I), for example, is indicated as E, this means that the compound contains substantially no Z-isomer; if a compound of formula (I), for example, is indicated as cis, this means that the compound contains substantially no trans isomer.

Some compounds according to formula (I) may also exist in their tautomeric form. It is intended that such forms, since they may exist although not explicitly shown in formula (I) above, are included within the scope of the present invention. It follows that one compound can exist in both stereoisomeric and tautomeric forms.

Pharmaceutically acceptable salts include acid addition salts and base addition salts. Such salts can be prepared by conventional methods, for example, by reacting the free acid or free base form with one or more equivalents of the corresponding base or acid, optionally in a solvent or medium in which the salt is insoluble, followed by removal of said solvent or specified medium using standard techniques (for example, in vacuo, freeze-drying or filtration). Salts can also be prepared by exchanging the counterion of a compound of the present invention in salt form with another counterion, for example using a suitable ion exchange resin.

The pharmaceutically acceptable salts mentioned above or below herein are intended to include the therapeutically active, non-toxic acid and base salt forms that the compounds of formula (I) and solvates thereof may form.

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

The compounds of formula (I) and their acid proton-containing solvates can also be converted into their non-toxic metal or amine salt forms by treatment with appropriate organic compounds.

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

The term solvate includes solvent addition forms, as well as their salts, which can form compounds of formula (I). Examples of such solvent addition forms are, for example, hydrates, alcoholates, and the like.

The compounds of the present invention obtained in the methods described below can be synthesized as mixtures of enantiomers, in particular racemic mixtures of enantiomers, which can be separated from each other following separation procedures known in the art. A method for separating the enantiomeric forms of compounds of formula (I) and their pharmaceutically acceptable salts and solvates involves liquid chromatography using a chiral stationary phase. Said pure stereochemically isomeric forms can also be prepared from the corresponding pure stereochemically isomeric forms of the corresponding starting materials, provided that the reaction proceeds stereospecifically. If a specific stereoisomer is desired, it is preferred that said compound be synthesized using stereospecific preparation methods. These methods will advantageously employ enantiomerically pure starting materials.

The present invention also covers isotopically labeled compounds of the present invention that are identical to those provided herein, except that one or more atoms are replaced by an atom with an atomic mass or mass number different from the atomic mass or mass number typically found in nature (or the most common found in nature).

All isotopes and isotopic mixtures of any particular atom or element, as defined herein, are considered within the scope of the compounds of the present invention, both naturally occurring and synthetically produced, both in naturally occurring isotopic composition and in isotopically enriched form. Exemplary isotopes that may be included in the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine and iodine, such as ² H, ³ H, ⁿ 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 ⁸² Br . Preferably, the radioactive isotope is selected from the group consisting of ² H, ³ H, 11 C and ¹⁸ F. More preferably, the radioactive isotope is ² H. In particular, deuterated compounds are intended to be included within the scope of the present invention.

Certain isotopically labeled compounds of the present invention (eg, ³ H and ¹⁴ C labeled) may be useful, for example, in substrate tissue distribution assays. The isotopes tritium ( ^3H ) and carbon-14 ( ^14C ) are useful because they are easy to obtain and can be detected. Additionally, substitution with heavier isotopes such as deuterium (i.e., H) may provide certain therapeutic benefits due to greater metabolic stability (e.g., increased in vivo half-life or reduced dose requirement) and may therefore be preferred in in some cases. Thus, in a particular embodiment of the present invention, R ² is selected from hydrogen or deuterium, in particular deuterium. In another embodiment, L ² may be >C( ² H) ₂ . Positron-active isotopes such as ¹⁵ O, ¹³ N, 11 C and ¹⁸ F are suitable for positron emission tomography (PET) studies. PET imaging in cancer finds utility in helping to localize and identify tumors, stage the disease, and determine appropriate treatment. Human cancer cells overexpress a variety of receptors or proteins that are potential molecular targets for a particular disease. Radiolabeled tags 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). Additionally, target-specific PET radiolabels can be used as biomarkers to study and evaluate pathology, for example by measuring target expression and response to treatment (Austin R. et al. Cancer Letters (2016), doi: 10.1016/j.canlet .2016.05.008).

In particular, the present invention relates to the compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the embodiments, which also excludes the following compound and its pharmaceutically acceptable addition salts:

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In particular, the present invention relates to the compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the embodiments, where the intermediates and compounds described in WO2017/112768 are excluded to the extent in which they are covered by the present invention.

In particular, the present invention relates to the compounds of formula (I) as defined herein, and to their tautomers and stereoisomeric forms, wherein:

(a) L ² is selected from the group consisting of >CR ^4a R ^4b and -CHR ^4a CHR ⁵ -; Where

L ² is bonded to the nitrogen atom of L ¹ , R ^4a is selected from the group consisting of hydrogen; -C(=O)NR ^7a R ^7b ; C _{b 4} alkyl, optionally substituted with a substituent selected from the group consisting of -OR ⁸ and -NR ^9a R ^9b ; and a C-linked 4-7 membered non-aromatic heterocyclyl containing at least one nitrogen atom;

R ⁵ is selected from the group consisting of hydrogen; -OR ⁶ ; -NR ^7a R ^7b and C _1-4 alkyl;

R ^4b is selected from the group consisting of hydrogen and methyl; or R ^4a and R ^4b together with the carbon atom to which they are attached form a C _3-5 cycloalkyl or a C 4-6 membered heterocyclyl containing an oxygen atom; wherein each of R ⁶ , R ^7a , R ^7b , R ⁸ , R ^9a and R ^9b is independently selected from the group consisting of hydrogen and C ₂₄ alkyl substituted with a substituent selected from the group consisting of -OR ¹¹ and -NR ^10a R ^10b ; wherein each of R ^10a , R ^10b and R ¹¹ is independently selected from the group consisting of hydrogen and C _1-4 alkyl; And

R ³ is selected from the group consisting of Ar, Het ¹ , Het ² and a 7-10 membered saturated spirocarbobicyclic system; or (b) L ² represents >CR ^4c R ^4d where each of R ^4c and R ^4d represents hydrogen; And

R ^12a .Ge^ _R i2b '' \ 12c

R ³ represents ' where each of R ^12a , R ^12b and R ^12c represents C _1-6 alkyl; or (c) --L ² -R ³ is C _1-6 alkyl, optionally substituted with one, two or three fluorine substituents; or (d) L ² represents O and R ³ represents -CH ₂ -Ar; or

(e) --L ² -R ³ represents ^R ' where

R ¹⁸ represents hydrogen; or

(f) --L ² -R ³ represents ' and where

Ar is phenyl, optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen and C _1-4 alkyl, optionally substituted with -OR ²⁶ ;

Het ¹ is a monocyclic heteroaryl selected from the group consisting of pyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl, 4- or 5-thiazolyl, isothiazolyl and isoxazolyl; or bicyclic heteroaryl selected from imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl; each of which may be optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -CN and C _1-4 alkyl, optionally substituted by a substituent selected from the group consisting of -CN, -OR ²⁶ and -NR ^27a R ^27b ; And

Het ² is a non-aromatic heterocyclyl selected from azetidinyl, pyrrolidinyl and piperidinyl; wherein each of R ²⁶ , R ^27a and R ^27b is independently selected from the group consisting of hydrogen and C _1-4 alkyl; and pharmaceutically acceptable salts thereof.

- 13 045364

In particular, the present invention relates to the compounds of formula (I) as defined herein, and to their tautomers and stereoisomeric forms, wherein

R ¹ represents CF3;

(a) L ² is >CR ^4a R ^4b ; Where

R ^4a is selected from the group consisting of hydrogen; -C(=O)NR ^7a R ^7b ; C ₁ . ₄ alkyl and C-linked 4-7 membered non-aromatic heterocyclyl containing at least one nitrogen atom; And

R ^4b is selected from the group consisting of hydrogen and methyl; wherein each of R ^7a and R ^7b is independently selected from the group consisting of hydrogen; C ₁ . ₄ alkyl and C ₂ . ₄ alkyl substituted with a substituent selected from the group consisting of -OR ¹¹ and -NR ^10a R ^10b ; wherein each of R ^10a , R ^10b and R ¹¹ is independently selected from the group consisting of hydrogen and C ₁ . ₄ alkyl; And

R ³ is selected from the group consisting of Ar, Het ¹ , Het ² and a 7-10 membered saturated spirocarbobicyclic system; or (b) L ² represents >CR ^4c R ^4d where R ^4c and R ^4d represent hydrogen; And

R ^12a ,Ge^. _R i2b\12c

R ³ represents ^R j where R ^12a , R ^12b and R ^12c represents C ₁ . ₆ alkyl; or (c) --L ² -R ³ is C ₁ . ₆ alkyl, optionally substituted with one, two or three substituents representing a fluorine atom; or (d) L ² represents O and R ³ represents -CH ₂ -Ar; or

(f) --L ² -R ³ is ^R R ¹⁸ is hydrogen; or

N— (g) --L2-R ³ represents

Ar represents phenyl, optionally substituted with a substituent representing a halogen atom;

Het ¹ is a monocyclic heteroaryl selected from the group consisting of pyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl and 4- or 5-thiazolyl; or bicyclic heteroaryl selected from imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl; each of which may be optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen and C _1-4 αlkylα, optionally substituted by a substituent selected from the group consisting of -CN, -OR ²⁶ and - NR ^27a R ^27b ; And

Het ² is a non-aromatic heterocyclyl selected from azetidinyl, pyrrolidinyl and piperidinyl; wherein each of R ²⁶ , R ^27a and R ^27b is independently selected from the group consisting of hydrogen and C ₁ . ₄ alkyl;

and pharmaceutically acceptable salts thereof.

In particular, the present invention relates to the compounds of formula (I) as defined herein, and to their tautomers and stereoisomeric forms, wherein

R ¹ represents CF ₃ ;

L ¹ is an N-linked 7-10 membered saturated spiroheterobicyclic system containing one or two N atoms selected from the group consisting of (a), (b), (c), (d), (e), ( f) and ⁽ g ^):

where a represents the position of binding to the thienopyrimidinyl heterocycle;

(a) L ² is >CH ₂ ; and R ³ is selected from the group consisting of Ar, Het ¹ and 7-10 membered on

- 14 045364 saturated spirocarbobicyclic system; or (b) --L2-R ³ is C ₁ . ₆ alkyl, optionally substituted with one, two or three substituents representing a fluorine atom; and where

Ar represents phenyl, optionally substituted with a substituent representing a halogen atom; And

Het ¹ is a monocyclic heteroaryl selected from the group consisting of 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl and 4- or 5-thiazolyl; or bicyclic heteroaryl selected from imidazopyridinyl, in particular imidazo[1,2a]pyridinyl; each of which may be optionally substituted with one or two substituents, each of which is independently selected from the group consisting of halogen and Aralkyl;

and pharmaceutically acceptable salts thereof.

In particular, the present invention relates to the compounds of formula (I) as defined herein, and to their tautomers and stereoisomeric forms, wherein

R ¹ represents CF ₃ ;

R ² represents hydrogen;

L ¹ is an N-linked 7-10 membered saturated spiroheterobicyclic system containing one or two N atoms selected from the group consisting of (a), (b), (c), (d), (e), ( f) and (g):

where a represents the binding position to the thienopyrimidinyl heterocycle;

(a) L ² is >CH ₂ ; and R ³ is selected from the group consisting of Ar, Het ¹ and a 7-10 membered saturated spirocarbobicyclic system; or (b) --L ² -R ³ is C ₁ . ₆ alkyl, optionally substituted with one, two or three substituents representing a fluorine atom; and where

Ar represents phenyl, optionally substituted with a substituent representing a halogen atom; And

Het ¹ is a monocyclic heteroaryl selected from the group consisting of 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl and imidazolyl; or bicyclic heteroaryl selected from imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl; each of which may be optionally substituted with one or two substituents, each of which is independently selected from the group consisting of halogen and C ₁ . ₄ alkyl;

and pharmaceutically acceptable salts thereof.

Another embodiment of the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, where one or more of the following limitations apply:

(a) R ¹ is CF3;

(b) R ² represents hydrogen;

(c) L ¹ is an N-linked 7-10 membered saturated spiroheterobicyclic system containing one or two N atoms selected from the group consisting of (a), (b), (c), (d), (e ), (f) and (g) defined herein;

(d) L ¹ is an N-linked 7-10 membered saturated spiroheterobicyclic system containing one or two N atoms selected from the group consisting of (a), (b), (c), (d), (e ) and (f) as defined herein;

(e) L ¹ is an N-linked 7-10 membered saturated spiroheterobicyclic system containing one or two N atoms selected from the group consisting of (c) and (e);

(f) L ² is >CH ₂ ;

(g) L ² is >CH ₂ ; and R ³ is selected from the group consisting of Ar, Het ¹ and a 7-10 membered saturated spirocarbobicyclic system;

(h) --L ² -R ³ selected from the group consisting of

- 15 045364

(i) Ar is phenyl, optionally substituted with one or two independently selected substituents representing a halogen atom;

(j) Ar represents phenyl, optionally substituted with one substituent representing a halogen atom;

(k) Ar is phenyl;

(l) Het ¹ is a monocyclic heteroaryl selected from the group consisting of pyrazolyl, imidazolyl, pyrrolyl, 4- or 5-thiazolyl, pyridyl, pyridazinyl, 4-, 5- or 6-pyrimidinyl and pyrazinyl; or is a bicyclic heteroaryl selected from imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl; each of which may be optionally substituted with one or two substituents, each independently selected from the group consisting of halogen and C1-4αalkylα, optionally substituted by a substituent selected from the group consisting of -CN, -OR ²⁵ and -NR ^26a R ^26b ; wherein each of R ²⁵ , R ^26a and R ^26b is independently selected from the group consisting of hydrogen and C1. ₄ alkyl;

(m) Het ¹ is a monocyclic heteroaryl selected from the group consisting of pyrazolyl, imidazolyl, pyrrolyl, 4- or 5-thiazolyl, pyridyl, pyridazinyl, 4-, 5- or 6-pyrimidinyl and pyrazinyl; or is a bicyclic heteroaryl selected from imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl; each of which may be optionally substituted with one or two substituents, each independently selected from the group consisting of halogen and C1-4αalkylα, optionally substituted by a substituent selected from the group consisting of -CN, -OR ²⁵ and -NR ^26a R ^26b ; wherein each of R ²⁵ , R ^26a and R ^26b is independently selected from the group consisting of hydrogen and C1. ₄ alkyl;

(n) Het ¹ is a monocyclic heteroaryl selected from the group consisting of pyrazolyl, imidazolyl, pyrrolyl, pyridazinyl, 4-, 5- or 6-pyrimidinyl and pyrazinyl; or is a bicyclic heteroaryl selected from imidazopyridinyl, in particular imidazo[1,2a]pyridinyl; each of which may be optionally substituted with one or two substituents, each of which is independently selected from the group consisting of halogen and C ₁ . ₄ alkyl;

(o) Het ¹ is a monocyclic heteroaryl selected from the group consisting of pyridazinyl, 4-, 5- or 6-pyrimidinyl and pyrazinyl, each of which may be optionally substituted with a substituent representing a halogen atom;

(p) Het ¹ is a bicyclic heteroaryl selected from imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl-6-yl or imidazo[1,2-a]pyridinyl-2-yl;

(q) The 7-10 membered saturated spirocarbobicyclic system is, in particular,

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein R ¹ represents CF3.

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof or any subgroup thereof specified in any of the other embodiments, wherein R ¹ is CF3 and wherein R ² is hydrogen.

- 16 045364

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof or any subgroup thereof specified in any of the other embodiments, wherein Ar is phenyl, optionally substituted according to any of the other embodiments.

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein L ² is bonded to the carbon atom L ¹ .

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L2-R ³ corresponds to (a).

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ corresponds to (b).

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ corresponds to (c).

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ corresponds to (d).

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ corresponds to (e).

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ corresponds to (f).

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ corresponds to (g).

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ corresponds to (a); (b), (d), (e) or (f); and R ^4a is other than hydrogen. In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ corresponds to (a) or ( f); and R ^4a is other than hydrogen. In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ corresponds to (a); and R ^4a is other than hydrogen.

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ corresponds to (a); and if L ² is bonded to the nitrogen atom of L ¹ , then R ^4a is other than hydrogen.

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ corresponds to (a) or ( f); and if L ² is bonded to the nitrogen atom of L ¹ , then R ^4a is other than hydrogen.

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ corresponds to (a), ( b), (d), (e) or (f); and if L ² is bonded to the nitrogen atom of L ¹ , then R ^4a is other than hydrogen.

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof or any subgroup thereof specified in any of the other embodiments, wherein L ¹ is

N I

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ corresponds to (a); R ³ represents Het ¹ or Het ² .

- 17 045364

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L2-R ³ corresponds to (a); R ³ represents Het ¹ .

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ corresponds to (a); R ³ represents Het ¹ ; and Het ¹ is azetidinyl, optionally substituted as defined in any other embodiment.

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ corresponds to (a); R ³ represents Het ¹ or Het ² ; Het ¹ is a monocyclic heteroaryl selected from the group consisting of pyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl and imidazolyl; each of which may be optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -CN, -OR ²⁴ and Yu^alkyl, optionally substituted by a substituent selected from the group consisting of -CN, -OR ²⁶ and -NR ^27a R ^27b ; And

Het ² is a non-aromatic heterocyclyl selected from the group consisting of azetidinyl, pyrrolidinyl and piperidinyl; each of which may be optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -CN, -OR ²⁴ and C ₁ . ₄ alkyl, optionally substituted with -OR ²⁶ .

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof or any subgroup thereof specified in any of the other embodiments, wherein Ar represents phenyl, which may be optionally substituted with one or two or three substituents, each independently selected from the group consisting of halogen, -CN, -OR ²⁴ and C ₁ . ₄ alkyl, optionally substituted -OR ²⁶ ;

Het ¹ is a monocyclic heteroaryl selected from the group consisting of pyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, 4- or 5-thiazolyl, isothiazolyl, thiadiazolyl and isoxazolyl; each of which may be optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -CN, -OR ²⁴ and C ₁ . ₄ alkyl, optionally substituted with a substituent selected from the group consisting of -CN, -OR ²⁶ and -NR ^27a R ^27b ; And

Het ² is a monocyclic non-aromatic heterocyclyl, optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -CN, -OR ²⁴ and C ₁ . ₄ alkyl, optionally substituted with -OR ²⁶ .

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof, or any subgroup thereof, as specified in any of the other embodiments, wherein

--L ² -R ³ matches (a), where

L ² is selected from the group consisting of >CR ^4a R ^4b and -CHR ^4a CHR ⁵ -; Where

L ² is connected to the nitrogen atom L ¹ ;

R ^4a is selected from the group consisting of -C(=O)NR ^7a R ^7b and a C-linked 4-7 membered non-aromatic heterocyclyl containing at least one nitrogen, oxygen or sulfur atom;

R ⁵ is selected from the group consisting of hydrogen; -OR ⁶ ;

-NR ^7a R ^7b ;

-C(=O)NR ^7a R ^7b ; C ₁ . ₄ alkyl, optionally substituted with a substituent selected from the group consisting of fluorine, -CN, -OR ⁸ and -NR ^9a R ^9b ;

R ^4b is selected from the group consisting of hydrogen and methyl; And

R ³ is selected from the group consisting of Ar, Het ¹ , Het ² and a 7-10 membered saturated spirocarbobicyclic system.

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof, or any subgroup thereof, as specified in any of the other embodiments, wherein

--L ² -R ³ matches (a), where

L ² represents >CR ^4a R ^4b ; Where

L ² is connected to the nitrogen atom L ¹ ;

R ^4a is selected from the group consisting of -C(=O)NR ^7a R ^7b and a C-linked 4-7 membered non-aromatic heterocyclyl containing at least one nitrogen, oxygen or sulfur atom;

R ^4b represents hydrogen; And

R ³ is selected from the group consisting of Ar, Het ¹ , Het ² and a 7-10 membered saturated spirocarbobicyclic system.

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof or any subgroup thereof specified in any of

- 18 045364 other embodiments, where

--L2-R ³ matches (a), where

L ² represents >CR ^4a R ^4b ; Where

L ² is connected to the nitrogen atom L ¹ ;

R ^4a is selected from the group consisting of -C(=O)NR ^7a R ^7b and a C-linked 4-7 membered non-aromatic heterocyclyl containing at least one nitrogen, oxygen or sulfur atom;

R ^4b represents hydrogen; And

R ³ is selected from the group consisting of Ar and Het ² .

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof, or any subgroup thereof, as specified in any of the other embodiments, wherein

- -L ² -R ³ corresponds to (a), where

L ² is selected from the group consisting of >CR ^4a R ^4b and -CHR ^4a CHR ⁵ -; Where

L ² is connected to the nitrogen atom L ¹ ;

R ^4a is -C(=O)NR ^7a R ^7b ;

R ⁵ is selected from the group consisting of hydrogen; -OR ⁶ ; -NR ^7a R ^7b ;

- C(=O)NR ^7a R ^7b ; C ₁ . ₄ alkyl, optionally substituted with a substituent selected from the group consisting of fluorine, -CN, -OR ⁸ and

- NR ^9a R ^9b ;

R ^4b is selected from the group consisting of hydrogen and methyl; And

R ³ is selected from the group consisting of Ar, Het ¹ , Her ² and a 7-10 membered saturated spirocarbobicyclic system.

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof, or any subgroup thereof, as specified in any of the other embodiments, wherein

- -L ² -R ³ corresponds to (a), where

L ² represents >CR ^4a R ^4b ; Where

L ² is connected to the nitrogen atom L ¹ ;

R ^4a is -C(=O)NR ^7a R ^7b ;

R ^4b represents hydrogen; And

R ³ is selected from the group consisting of Ar, Het ¹ , Het ² and a 7-10 membered saturated spirocarbobicyclic system.

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof, or any subgroup thereof, as specified in any of the other embodiments, wherein

- -L ² -R ³ corresponds to (a), where

L ² represents >CR ^4a R ^4b ; Where

L ² is connected to the nitrogen atom L ¹ ;

R ^4a is -C(=O)NR ^7a R ^7b ;

R ^4b represents hydrogen; And

R ³ is selected from the group consisting of Ar and Het ² .

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof, or any subgroup thereof, as specified in any of the other embodiments, wherein

- -L ² -R ³ corresponds to (a), where

L ² is selected from the group consisting of >CR ^4a R ^4b and -CHR ^4a CHR ⁵ -; Where

L ² is connected to the nitrogen atom L ¹ ;

R ^4a is a C-linked 4-7 membered non-aromatic heterocyclyl containing at least one nitrogen, oxygen or sulfur atom;

R ⁵ is selected from the group consisting of hydrogen; -OR ⁶ ; -NR ^7a R ^7b ;

- C(=O)NR ^7a R ^7b ; C ₁ . ₄ alkyl, optionally substituted with a substituent selected from the group consisting of fluorine, -CN, -OR ⁸ and

- NR ^9a R ^9b ;

R ^4b is selected from the group consisting of hydrogen and methyl; And

R ³ is selected from the group consisting of Ar, Het ¹ , Het ² and a 7-10 membered saturated spirocarbobicyclic system.

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof, or any subgroup thereof, as specified in any of the other embodiments, wherein

- -L ² -R ³ corresponds to (a), where

L ² represents >CR ^4a R ^4b ; Where

L ² is connected to the nitrogen atom L ¹ ;

- 19 045364

R ^4a is a C-linked 4-7 membered non-aromatic heterocyclyl containing at least one nitrogen, oxygen or sulfur atom;

R ^4b represents hydrogen; And

R ³ is selected from the group consisting of Ar, Het ¹ , Het ² and a 7-10 membered saturated spirocarbobicyclic system.

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof, or any subgroup thereof, as specified in any of the other embodiments, wherein

--L ² -R ³ matches (a), where

L ² represents >CR ^4a R ^4b ; Where

L ² is connected to the nitrogen atom L ¹ ;

R ^4a is a C-linked 4-7 membered non-aromatic heterocyclyl containing at least one nitrogen, oxygen or sulfur atom;

R ^4b represents hydrogen; And

R ³ is selected from the group consisting of Ar and Het ² .

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein L ² is bonded to the nitrogen atom L ¹ .

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ is selected from the group consisting of

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof or any subgroup thereof specified in any of the other embodiments, wherein --L ² -R ³ is selected from the group consisting of

where R ¹⁸ represents hydrogen or methyl.

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof, or any subgroup thereof, as specified in any of the other embodiments, wherein

L ¹ is an N-linked 7-10 membered saturated spiroheterobicyclic system containing one or two N atoms selected from the group consisting of (a), (b), (c), (d), (e), ( f), (g), (h) and (i):

- 20 045364

where a denotes the binding position with the thienopyrimidinyl heterocycle.

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof or any subgroup thereof specified in any of the other embodiments, wherein Het ² is a monocyclic heterocyclyl, optionally substituted with one, two or three substituents described in other embodiments.

In one embodiment, the present invention relates to such compounds of formula (I) and their pharmaceutically acceptable salts or any subgroup thereof specified in any of the other embodiments, wherein Het ² is a non-aromatic heterocyclyl selected from azetidinyl, oxetanyl, tetrahydrofuranyl, piperidinyl, tetrahydropyranyl, tetrahydro2H-thiopyranyl-1,1-dioxide, α-,Ά / ^no /-AND', each of which is optionally substituted with one, two or three substituents described in other embodiments.

In one embodiment, the present invention relates to such compounds of formula (I) and pharmaceutically acceptable salts thereof or any subgroup thereof specified in any of the other embodiments, wherein Het ² is a non-aromatic heterocyclyl selected from

each of which is optionally substituted with one, two or three substituents described in other embodiments.

Specific compounds of formula (I) are compounds 82, 84, 273 and 274, including their stereoisomeric forms, pharmaceutically acceptable salts, in particular their hydrochloride salts.

Specific compounds of formula (I) are compounds 82, 84, 273 and 274.

In one embodiment, the compound of formula (I) is selected from the group consisting of any of the exemplary compounds and their free bases, pharmaceutically acceptable addition salts.

It is intended that all possible combinations of the above embodiments fall within the scope of the present invention.

- 21 045364

Methods for preparing compounds of formula (I).

In this section, as in all other sections, unless the context indicates otherwise, references to claim (I) also include all other subclauses and examples thereof, as defined herein.

General methods for preparing some representative examples of compounds of formula (I) are described herein and in the specific examples, and are generally prepared from starting materials either commercially available or prepared by standard synthetic methods commonly used by those skilled in the art. technology. It is intended that the following diagrams only represent examples of the present invention and are not intended to limit the present invention in any way.

Alternatively, the compounds of the present invention can also be prepared using reaction protocols similar to those described in the general schemes below, in combination with standard synthetic methods commonly used by those skilled in the art of organic chemistry.

One skilled in the art will appreciate that in the reactions described in the schemes, although not always explicitly shown, protection of reactive functional groups (eg hydroxy, amino or carboxy groups) may be necessary if they are needed in the final product in order to to avoid their unwanted participation in reactions. For example, in Scheme 1, the NH fragment on L ¹ of a N-linked 7-10 membered saturated spiroheterobicyclic system containing one or two N atoms can be protected using a tert-butoxycarbonyl protecting group. In general, traditional protecting groups can be used in accordance with standard practice. The protecting groups can be removed at a suitable subsequent stage using methods known in the art. This is illustrated with specific examples.

One skilled in the art will appreciate that in the reactions described in the schemes, it may be advisable or necessary to conduct the reaction under an inert atmosphere, such as, for example, an atmosphere of N2 gas.

It will be apparent to one skilled in the art that it may be necessary to cool the reaction mixture before processing the reaction product (referring to a number of procedures necessary to isolate and purify the chemical reaction product(s), such as, for example, quenching, column chromatography, extraction) .

One skilled in the art will appreciate that heating the reaction mixture while stirring can increase the yield of the reaction. In some reactions, microwave heating can be used instead of traditional heating to reduce the overall reaction time.

One skilled in the art will appreciate that another sequence of chemical reactions shown in the diagrams below may also result in the desired compound of formula (I).

One skilled in the art will appreciate that the intermediates and final compounds shown in the diagrams below can be further functionalized in accordance with methods well known to one skilled in the art. The intermediates and compounds described herein can be isolated in free form or as a salt.

Scheme 1.

In general, compounds of formula (I), wherein all variables are defined within the scope of the present invention, can be prepared in accordance with the following reaction scheme 1. In scheme 1, X—-bb means L ¹ as a 7-10 membered saturated spiroheterobicyclic system containing two N atoms, and which is N-linked to a thienopyrimidinyl heterocycle, each of LG ¹ and LG ² represents a suitable leaving group, such as, for example, halogen or methanesulfonyl; PG ¹ represents a suitable protecting group such as, for example, tert-butyloxycarbonyl; R ^3a -PG ² is R ³ as defined in formula (I) with an appropriate protecting group, such as, for example, tert-butyloxycarbonyl, if the R ³ substituent contains an amino group. X in formula (XI) represents CH or N (in formula (XI), L2 may be bonded to a carbon or an N atom). All other variables in Scheme 1 are defined within the scope of the present invention.

- 22 045364

In Scheme 1, the following reaction conditions are applied:

1: at a suitable temperature, such as in the range from b.t. up to 90°C, in the presence of a suitable base, such as, for example, diisopropylethylamine, in a suitable solvent, such as, for example, acetonitrile, or isopropanol, or ethanol;

2: at a suitable temperature ranging from, for example, 0°C to room temperature, in the presence of suitable digestion conditions, such as, for example, an acid such as HCl or trifluoroacetic acid, in a suitable solvent such as acetonitrile or dichloromethane if PG ¹ is tert-butyloxycarbonyl.

Alternatively, at a suitable temperature, such as, for example, room temperature, in a suitable solvent such as acetic acid.

3: at a suitable temperature, such as, for example, room temperature or reflux temperature, in the presence of a suitable base, such as, for example, potassium carbonate or 1,8diazabicyclo[5.4.0]undec-7-ene, in a suitable solvent such such as acetonitrile or DMSO;

4: at a suitable temperature, such as, for example, room temperature or 90°C, in the presence of a suitable base, such as, for example, potassium carbonate or 1,8-diazabicyclo[5.4.0]undec-7ene, in a suitable solvent such such as acetonitrile or DMSO;

5: at a suitable temperature ranging from, for example, 0°C to room temperature, in the presence of suitable digestion conditions, such as, for example, an acid such as HCl or trifluoroacetic acid, in a suitable solvent such as acetonitrile or dichloromethane if PG ² is tert-butyloxycarbonyl;

6: at a suitable temperature, such as, for example, room temperature, then in the presence of a suitable base, such as, for example, trimethylamine, or a suitable acid, such as, for example, acetic acid, in a suitable solvent, such as, for example, anhydrous dichloromethane, dichloroethane or tetrahydropyran;

7: at a suitable temperature, for example room temperature, in the presence of a suitable reducing agent, such as, for example, NaBH(OAc) ₃ , in a suitable solvent such as dichloromethane, dichloroethane or tetrahydropyran; to obtain a compound of formula (I), wherein L ¹ is an N-linked 7-10 membered saturated spiroheterobicyclic system containing two N atoms, and L ² is CH ₂ .

Steps 6 and 7 can be easily carried out through a single reactor procedure.

Alternatively, steps 6 and 7 can be carried out in the presence of a suitable acid, such as, for example, acetic acid, a suitable catalyst, such as platinum oxide, in a suitable solvent, such as, for example, ethanol, at a suitable temperature, such as, for example, , 60°C;

8: at a suitable temperature, such as, for example, 90°C, in the presence of a suitable base, such as, for example, diisopropylethylamine, in a suitable solvent, such as, for example, acetonitrile or isopropanol. In step 8, in the reagent of formula (XI), X is CH or N, and L ² and R ³ are defined within the scope of the present invention. The reagents of formula (XI) are either commercially available or can be prepared by methods known to one skilled in the art from commercially available starting materials, for example, by appropriate deprotection/deprotection and interconversion steps.

- 23 045364 national groups, from starting materials such as 2-azaspiro[3.3]heptan-6-ol (CAS[1256352-97-2]).

Scheme 2.

Intermediates of formula (II) wherein R ² is methyl can be prepared according to the following reaction scheme 2, where LG ¹ is a suitable leaving group such as, for example, halogen or methanesulfonyl. All other variables in Scheme 2 are defined within the scope of the present invention.

In Scheme 2, the following reaction conditions are applied:

1: at a suitable temperature, such as, for example, reflux temperature, in the presence of acetic anhydride and a suitable base, such as, for example, trimethylamine, in a suitable solvent, such as, for example, toluene;

2: at a suitable temperature, such as, for example, reflux temperature, in the presence of a suitable base, such as potassium hydroxide, in a suitable solvent, such as, for example, ethanol;

3: under suitable reaction conditions to form a leaving group such as chlorine, for example by reacting with phosphorus trichloride at a suitable temperature such as 110°C.

Scheme 3.

In general, compounds of formula (Ia), where the variables are defined within the scope of the present invention, but where L ² is limited by L ^2a (variants that can be prepared using this scheme), can be prepared in accordance with the following reaction scheme 3. All others the variables in Scheme 3 are defined within the scope of the present invention or as defined above.

In Scheme 3 the following reaction conditions are applied:

- 24 045364

1: at a suitable temperature, such as, for example, room temperature or 45°C, in the presence of titanium(IV) ethoxide or titanium(IV) isopropoxide, in a suitable solvent such as, for example, tetrahydropyran, dichloroethane or a mixture of dichloroethane and methanol ;

Alternatively, at a suitable temperature, such as, for example, room temperature, with or without a suitable acid, such as, for example, trifluoroacetic acid, in a suitable solvent, such as, for example, tetrahydropyran;

2: at a suitable temperature, such as, for example, room temperature, in the presence of a suitable reducing agent, such as, for example, sodium borohydride, sodium triacetoxyborohydride or sodium cyanoborohydride, in a suitable solvent, such as, for example, tetrahydropyran, dichloroethane or a mixture of dichloroethane and methanol ;

Stages 1 and 2 can be carried out through a procedure carried out in one reactor.

Scheme 4.

In general, compounds of formula (Ib) wherein R ^4a is limited to R ^4a1 being a C _1-4 alkyl or C 4-7 membered non-aromatic heterocyclyl containing at least one nitrogen, oxygen or sulfur atom can be prepared in according to the following reaction scheme 4. In scheme 4, halogen means bromine, chlorine or iodine. All other variables in Scheme 4 are defined within the scope of the present invention or as defined above.

In Scheme 4 the following reaction conditions are applied:

1: at a suitable temperature, such as, for example, room temperature or 45°C, in the presence of titanium(IV) ethoxide or titanium(IV) isopropoxide, in a suitable solvent, such as, for example, tetrahydropyran;

2: at a suitable temperature ranging from 0°C to room temperature, in a suitable solvent such as, for example, tetrahydrofuran.

Stages 1 and 2 can be carried out through a procedure carried out in one reactor.

Scheme 5.

In general, compounds of formula (Ic) wherein R ³ is limited to R ^3c being

can be prepared according to the following Reaction Scheme 5. All other variables in Scheme 5 are defined within the scope of the present invention or as defined above. In Scheme 5, L ² is bonded to the NL ¹ atom.

In Scheme 5 the following reaction conditions are applied:

1: at a suitable temperature, such as, for example, room temperature, in the presence of a suitable acid coupling agent, such as, for example, 1-[bis-(dimethylamino)methylene]-1Hbenzotriazolium hexafluorophosphate (1-)3-oxide (HBTU) or 1-[bis-(dimethylamino)methylene]-1H-1,2,3triazolo[4,5-b]pyridinium-3-oxide hexafluorophosphate (HATU), in the presence of a suitable base,

- 25 045364 such as, for example, N-ethyl-N-(1-methylethyl)-2-propanamine (DIPEA), in a suitable solvent such as N,N-dimethylformamide (DMF);

Scheme 6.

In general, compounds of formula (Id) wherein L ² is limited to SO2 can be prepared according to the following reaction scheme 6. All other variables in scheme 6 are defined within the scope of the present invention or as defined above. In Scheme 6, L ² (>SO2 in Scheme 6) is bonded to the NL ¹ atom.

In Scheme 6, the following reaction conditions are applied:

1: at a suitable temperature, for example room temperature, in the presence of a suitable base, such as, for example, potassium carbonate, in a suitable solvent, such as, for example, acetonitrile.

Scheme 7.

In general, compounds of formula (Ie) and (If) can be prepared according to the following reaction scheme 7. In both (Ie) and (If), part of the L ² molecule is bonded to the nitrogen atom L ¹ . All other variables are defined within the scope of the present invention or as defined above.

In Scheme 7 the following reaction conditions are applied:

1: at a suitable temperature, such as, for example, 60°C, in a suitable solvent, such as, for example, ethanol.

It should be understood that if appropriate functional groups are present, the compounds of various formulas or any intermediates used to prepare them can be further derivatized by one or more standard synthetic methods using condensation, substitution, oxidation, reduction or cleavage reactions. Specific substitution approaches include traditional alkylation, arylation, heteroarylation, acylation, sulfonylation, halogenation, nitration, formylation and coupling procedures.

The compounds of formula (I) can be synthesized as racemic mixtures of enantiomers, which can be separated from each other following separation procedures known in the art. Racemic compounds of formula (I) containing a basic nitrogen atom can be converted into the corresponding diastereomeric salt forms by reaction with a suitable chiral acid. These diastereomeric salt forms are then separated, for example by selective or fractional crystallization, and the enantiomers are isolated from there using alkali. An alternative method for separating the enantiomeric forms of compounds of formula (I) involves liquid chromatography using a chiral stationary phase. Said pure stereochemically isomeric forms can also be prepared from the corresponding pure stereochemically isomeric forms of the corresponding starting materials, provided that the reaction proceeds stereospecifically.

When preparing the compounds of the present invention, it may be necessary to protect the remote functionality (eg, primary or secondary amine) of the intermediates. The need to provide such protection will vary depending on the nature of the individual functional group and the conditions of the preparation methods. Suitable amino protecting groups (NHPg) include acetyl, trifluoroacetyl, tert-butoxycarbonyl (Boc), benzyloxycarbonyl (CBz) and 9fluorenylmethyleneoxycarbonyl (Fmoc). The need for such protection will be readily determined by one skilled in the art. For a general description of protecting groups and their applications, see T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 4th ed., Wiley, Hoboken, New Jersey, 2007.

- 26 045364

Pharmacology.

The compounds of the present invention have been found to block the interaction of menin with MLL proteins and oncogenic MLL-based fusion proteins. Therefore, the compounds of the present invention and pharmaceutical compositions containing these compounds may be useful for the treatment or prevention, in particular the treatment, of diseases such as cancer, myelodysplastic syndrome (MDS) and diabetes.

In particular, the compounds of the present invention and pharmaceutical compositions based thereon may be useful in the treatment or prevention of cancer. In one embodiment, the types of cancer for which treatment with the menin-MLL inhibitors of the present invention may be appropriate include types of leukemia, myeloma, or cancer that is a solid tumor (eg, prostate cancer, lung cancer, breast cancer , pancreatic cancer, colon cancer, liver cancer, melanoma and glioblastoma, etc.). In some embodiments, the leukemia types include acute leukemia types, chronic leukemia types, myeloid leukemia types, myelogenous leukemia types, lymphoblastic leukemia types, lymphocytic leukemia types, acute myelogenous leukemia (AML) types, chronic myelogenous leukemia (CML) types, acute lymphoblastic leukemia types leukemia (ALL), types of chronic lymphocytic leukemia (CLL), types of T-cell prolymphocytic leukemia (T-PLL), leukemia of large granular lymphocytes, hairy cell leukemia (HCL), types of leukemia with MLL rearrangement, types of leukemia associated with MLL- PTD, types of leukemia with MLL amplification, types of MLL-positive leukemia, types of leukemia characterized by HOX/MEIS1 gene expression profiles, etc.

Therefore, the present invention relates to the compounds of formula (I), their tautomers and stereoisomeric forms, as well as their pharmaceutically acceptable salts and solvates for use as a drug.

The present invention also relates to the use of a compound of formula (I), a tautomer or stereoisomeric form thereof, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to the present invention for the manufacture of a medicinal product.

The present invention also relates to a compound of formula (I), a tautomer or stereoisomeric form thereof, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition in accordance with the present invention for use in the treatment, prevention, mitigation, control or reduction of the risk of developing disorders associated with the interaction of menin with MLL proteins and oncogenic MLL-based fusion proteins in a mammal, including humans, the treatment or prevention of which is dependent on or facilitated by blocking the interaction of menin with MLL proteins and oncogenic MLL-based fusion proteins.

Also, the present invention relates to the use of a compound of formula (I), a tautomer or stereoisomeric form thereof, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition in accordance with the present invention for the manufacture of a medicament for the treatment, prevention, amelioration, control or reduction of the risk of developing disorders associated with the interaction of menin with MLL proteins and oncogenic MLL-based fusion proteins in a mammal, including humans, the treatment or prevention of which is dependent on or facilitated by blocking the interaction of menin with MLL proteins and oncogenic MLL-based fusion proteins.

The present invention also provides a compound of formula (I), a tautomer or stereoisomeric form thereof, or a pharmaceutically acceptable salt or solvate thereof for use in the treatment or prevention of any of the diseases mentioned herein above.

The present invention also provides a compound of formula (I), a tautomer or stereoisomeric form thereof, or a pharmaceutically acceptable salt or solvate thereof for use in the treatment or prevention of any of the diseases mentioned herein above.

The present invention also relates to the use of a compound of formula (I), a tautomer or stereoisomeric form thereof, or a pharmaceutically acceptable salt or solvate thereof for the manufacture of a medicament for the treatment or prevention of any of the disease states mentioned herein above.

The compounds of the present invention can be administered to mammals, preferably humans, for the treatment or prevention of any of the diseases mentioned above herein.

In view of the usefulness of the compounds of formula (I), their tautomers and stereoisomeric forms, and their pharmaceutically acceptable salts and solvates, a method is provided for the treatment of warm-blooded animals, including humans, suffering from any of the diseases mentioned above herein.

Said method includes introducing, i.e. systemic or local administration, preferably oral administration, of a therapeutically effective amount of a compound of formula (I), a tautomer or stereoisomeric form thereof, or a pharmaceutically acceptable salt or solvate thereof, to warm-blooded animals, including humans.

Therefore, the present invention also relates to a method of treating or preventing any of the diseases mentioned herein above, comprising administering a therapeutic

- 27045364 a commercially effective amount of a compound according to the present invention to a patient in need thereof.

One skilled in the art will appreciate that a therapeutically effective amount of the compounds of the present invention is an amount sufficient to exhibit therapeutic activity, and that this amount varies inter alias depending on the type of disease, the concentration of the compound in the therapeutic composition, and the condition of the patient. In general, the amount of a compound of the present invention to be administered as a therapeutic agent for the treatment of the disorders described herein will be determined on a case-by-case basis by the attending physician.

Those skilled in the treatment of such diseases will be able to determine the effective therapeutic daily amount from the test results presented below in this document. An effective daily therapeutic amount will be from about 0.005 mg/kg to 100 mg/kg, in particular from 0.005 mg/kg to 50 mg/kg, in particular from 0.01 mg/kg to 50 mg/kg body weight, more specifically from 0.01 mg/kg to 25 mg/kg body weight, preferably from about 0.01 mg/kg to about 15 mg/kg, more preferably from about 0.01 mg/kg to about 10 mg/kg, even more preferably from about 0.01 mg/kg to about 1 mg/kg, most preferably from about 0.05 mg/kg to about 1 mg/kg body weight. The specific effective daily therapeutic amount may be 1 mg/kg body weight, 2 mg/kg body weight, 4 mg/kg body weight, or 8 mg/kg body weight. The amount of a compound of the present invention, also referred to herein as the active ingredient, that is necessary to achieve a therapeutic effect may vary on a case-by-case basis, for example, for a particular compound, route of administration, age and condition of the recipient, and a particular disorder or condition. disease to be treated. The method of treatment may also include administering the active ingredient according to a schedule of one to four administrations per day. In these treatments, the compounds of the present invention are preferably formulated prior to administration. As described herein below, suitable pharmaceutical compositions are prepared by known procedures using well known and publicly available ingredients.

The present invention also provides compositions for the prevention or treatment of the disorders described herein. Said compositions contain a therapeutically effective amount of a compound of formula (I), a tautomer or stereoisomeric form thereof, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier or diluent.

Although the active ingredient can be administered separately, it is preferable if it is presented in the form of a pharmaceutical composition. Therefore, the present invention further provides a pharmaceutical composition containing a compound of the present invention together with a pharmaceutically acceptable carrier or diluent. The carrier or diluent must be acceptable in terms of compatibility with the other ingredients of the composition and must not be harmful to patients receiving it.

The pharmaceutical compositions of the present invention can be prepared by any methods well known in the art of pharmacy, for example by methods such as those described in Gennaro et al. Remington's Pharmaceutical Sciences ( ^18th ed., Mack Publishing Company, 1990, see especially Part 8: Pharmaceutical preparations and their Manufacture). A therapeutically effective amount of a particular compound, in base or salt form, as the active ingredient is combined in a uniform mixture with a pharmaceutically acceptable carrier, which may take a number of forms depending on the form of preparation required for administration. It is desirable that these pharmaceutical compositions be in unit dosage form, preferably suitable for systemic administration, such as oral, transdermal or parenteral administration; or for topical administration, such as by inhalation, nasal spray, eye drops, or by cream, gel, shampoo, or the like. For example, when preparing the compositions in the form of a dosage form for oral administration, any conventional pharmaceutical vehicles can be used, such as, for example, water, glycols, oils, alcohols, etc., in the case of liquid preparations for oral administration, such as suspensions, syrups , tinctures and solutions; or solid carriers such as starches, sugars, kaolin, lubricants, binders, disintegrants and the like, in the case of powders, pills, capsules and tablets. Due to their ease of administration, tablets and capsules are the most advantageous unit dosage forms for oral administration, in which case solid pharmaceutical carriers are of course used. For compositions for parenteral administration, the carrier will generally comprise sterile water, at least in large part, although other ingredients may be included, for example, to aid solubility. For example, injectable solutions can be prepared in which the carrier contains saline solution, glucose solution or a mixture of saline solution and glucose solution. Injectable suspensions can also be prepared, in which case suitable liquid carriers, suspending agents and the like can be used. In compositions suitable for transdermal administration, the carrier optionally contains a penetration enhancing agent and/or a suitable wetting agent, optionally

- 28 045364 combinations with suitable additives of any nature in minimal proportions, while the additives do not have any significant harmful effects on the skin. These additives may facilitate administration into the skin and/or may be useful in preparing the desired compositions. These compositions can be administered in various ways, for example, through a transdermal patch, by spot application, or as an ointment.

It is particularly advantageous to formulate the above-mentioned pharmaceutical compositions in unit dosage form for ease of administration and uniformity of dosage. Unit dosage forms, as used herein and in the claims, refer to physically discrete units suitable as dosage units, each unit containing a predetermined amount of the active ingredient calculated to produce the desired therapeutic effect, together with the required pharmaceutical carrier. Examples of such unit dosage forms are tablets (including divisible or film-coated tablets), capsules, pills, powder packets, wafers, injectable solutions or suspensions, heaped teaspoons, heaped tablespoons, and the like. ., as well as their individual multiples.

The compounds of the present invention can be used for systemic administration, such as oral, transdermal or parenteral administration; or for topical administration, such as by inhalation, nasal spray, eye drops, or by cream, gel, shampoo or the like. The compounds are preferably administered orally. The exact dosage and frequency of administration will depend on the particular compound of formula (I) used, the particular condition being treated, the severity of the condition being treated, the age, weight, sex, severity of the disorder and the general physical condition of the particular patient, as well as other drug treatment, which an individual may receive, as is well known to those skilled in the art. Moreover, it will be appreciated that said effective daily amount may be reduced or increased depending on the response of the subject being treated and/or depending on the judgment of the physician prescribing the compounds of the present invention.

The compounds of the present invention can be administered alone or in combination with one or more additional therapeutic agents. Combination therapy includes administration of a single dosage pharmaceutical formulation that contains a compound of the present invention and one or more additional therapeutic agents, as well as administration of a compound of the present invention and each additional therapeutic agent in its own separate dosage pharmaceutical composition. For example, a compound of the present invention and a therapeutic agent may be administered to a patient together in a single oral dosage composition, such as a tablet or capsule, or each agent may be administered separately in oral dosage formulations.

Therefore, an embodiment of the present invention relates to a product containing, as a first active ingredient, a compound according to the present invention, and as a further active ingredient, one or more anticancer agents, in the form of a combination preparation for simultaneous, separate or sequential use in the treatment of patients suffering from cancer.

One or more other medicinal agents and a compound of the present invention may be administered simultaneously (eg, in separate or single compositions) or sequentially in a random order. In the latter case, two or more compounds will be administered over a period and in an amount and manner that are sufficient to ensure that an advantageous or synergistic effect is achieved. It should be understood that the preferred route and order of administration and the corresponding dosage levels and schedules for each component of the combination will depend on the particular other medicinal agent and compound of the present invention to be administered, their route of administration, the particular condition, in particular the tumor, to be treated, and specific host to be treated. The optimal route and order of administration, as well as dosage levels and schedules, can be readily determined by those skilled in the art using standard techniques and taking into account the information set forth herein.

The weight ratio of the compound of the present invention and one anticancer agent or several other anticancer agents administered in combination can be determined by one skilled in the art. The ratio, precise dosage, and frequency of administration depend on the particular compound of the present invention and other anticancer agent(s) used, the specific condition being treated, the severity of the condition being treated, age, body weight, sex, diet, the time of administration and the general physical condition of the particular patient, the route of administration, as well as other drug treatments that the individual may be receiving, as is well known to those skilled in the art. Moreover, it is obvious that the effective daily amount can be reduced or

- 29 045364 increase depending on the response of the subject being treated and/or depending on the assessment of the attending physician prescribing the compounds of the present invention. The specific weight ratio for a given compound of formula (I) and the other anticancer agent may be in the range of 1/10 to 10/1, more particularly 1/5 to 5/1, even more particularly 1/3 to 3/ 1.

The following examples further illustrate the present invention.

Examples

Some methods for preparing the compounds of the present invention are illustrated in the following examples. Unless otherwise stated, all starting materials were obtained from commercial suppliers and used without further purification.

As used herein, the terms: ACN or MeCN means acetonitrile, DCM means dichloromethane, DIPEA means N,N-diisopropylethylamine, DIPE or DiPE means diisopropyl ether, h means hour(s), min means minute(s), DMF means dimethylformamide, DSC means differential scanning calorimetry, TEA or Et ₃ N means triethylamine, Et2O means diethyl ether, EtOAc or EA means ethyl acetate, EtOH means ethanol, HPLC means high performance liquid chromatography, iPrOH means isopropyl alcohol, LC/MS means liquid chromatography/mass spectrometry , MeOH means methanol, NMR means nuclear magnetic resonance, r.t. or RT means room temperature, SFC means supercritical fluid chromatography, OR means optical rotation, sat. aq. means saturated aqueous. AcCl means acetyl chloride, AcOH or HOAc means acetic acid, BOC or Boc means tert-butyloxycarbonyl, Celite® means diatomaceous earth, CH3COONH4 means ammonium acetate, COMU® means (1-cyano-2-ethoxy-2oxoethylideneaminooxy)dimethylaminomorpholinocarbenium hexafluorophosphate, CO2 means dioxide carbon, DCE means dichloroethane, DMAP means dimethylaminopyridine, DMSO means dimethyl sulfoxide, DBU means 1,8-diazabicyclo[5.4.0]undecene-7, EDCI.HCl means 1-(3dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, ee means enantiomeric excess, eq. or equivalent means equivalent(s), EtMgBr means bromoethylmagnesium, EtOAc means diethyl ether, EtOAc means ethyl acetate, Et ₃ N or TEA means triethylamine, EtOH means ethanol, h means hour(s), HATU means O-(7-azabenzotriazole-1- yl)-N,N,N,Ntetramethyluronium hexafluorophosphate, HCl means hydrochloric acid, HOBT means Nhydroxybenzotrizole monohydrate, H ₂ O means water, iPrMgCl means isopropyl magnesium chloride, iPrNH ₂ means isopropylamine, K2CO3 means potassium carbonate, Me-THF means 2methyltetrahydrofuran, MeMgBr or CH ₃ MgBr means methyl magnesium bromide, MeOH means methanol, MgSO ₄ means magnesium sulfate, min means minute(s), NaBH(OAc) ₃ means sodium triacetoxyborohydride, NaBH ₃ CN means sodium cyanoborohydride, Na ₂ CO ₃ means sodium carbonate, NaH means sodium hydride, NaHCO ₃ means sodium bicarbonate, NaOH means potassium hydroxide, Na ₂ SO ₄ means sodium sulfate, NH ₄ Cl means ammonium chloride, NH4HCO3 means ammonium bicarbonate, NH4OH means 30% aqueous ammonia solution, qty. or quantities. means quantitative, R _t means retention time, SFC means supercritical fluid chromatography, T means temperature, TBAF means tetrabutylammonium fluoride, TBDMS or SMDBT means tert-butyldimethylsilyl, TFA or CF3COOH means trifluoroacetic acid, THF means tetrahydrofuran, Ti(OEt) ₄ means ethoxide titanium, Ti(OiPr) ₄ means titanium isopropoxide, vol. means volume, F3C or CF3 means trifluoromethyl, HBTU means 1-[bis(dimethylamino)methylene]-1 H-benzotriazolium hexafluorophosphate(1-)3-oxide.

One skilled in the art will appreciate that compounds synthesized using these protocols may exist as a solvate, such as a hydrate, and/or may contain residual solvent or minor impurities. Compounds isolated in salt form may have integer stoichiometry, i.e. mono- or disols, or intermediate stoichiometry.

The stereochemical configuration for centers in some compounds may be designated R or S if the mixture(s) are separated; For some compounds, the stereochemical configuration at the specified centers was designated as *R (eluted first from the column in the case of the column conditions described in the synthesis protocol and if only one stereocenter is present) or *S (eluted second from the column in the case of the conditions described in the synthesis protocol for the column). columns and if only one stereocenter is present) if the absolute stereochemistry is not determined (even if the bonds are drawn stereospecifically), although the compound itself has been isolated as a single stereoisomer and is enantiomerically pure.

- 30 045364

For example, it will be appreciated that compound 179 is

Compounds characterized by two stereocenters and in which only one stereochemical configuration of one stereocenter is indicated by * (e.g., *R or *S) (see, e.g., compound 186 or 281) follow a similar rule to the one above. This means that the absolute stereochemical configuration of the stereocenter indicated by * is not determined (even if the bonds are drawn stereospecifically), although the compound is enantiomerically pure at the center indicated.

For compounds such as 188, 189, 190, 191, 235, 236, 237, and 238, where the stereochemical configuration of the two stereocenters is indicated by * (e.g., *R or *S), the absolute stereochemistry of the stereocenters is not determined (even if the bonds are drawn stereospecific), although the compound itself has been isolated as a single stereoisomer and is enantiomerically pure. In this case, the configuration of the first stereocenter is independent of the configuration of the second stereocenter in the same compound.

For example, for connection 188

this means that the connection is

The paragraphs above about stereochemical configurations also apply to intermediates.

The term enantiomerically pure as used herein means that the product contains at least 80% by weight of one enantiomer and 20% by weight or less of the other enantiomer. Preferably, the product contains at least 90% by weight of one enantiomer and 10% by weight or less 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 1% or less of the other enantiomer.

- 31 045364

If an intermediate or compound in the experimental section below is listed as HCl salt, HCOOH salt, or TFA salt without specifying the number of HCl or TFA equivalents, it means that the number of HCl or TFA equivalents was not determined.

One skilled in the art will appreciate that, even if not explicitly stated in the experimental protocols below, typically, after purification by column chromatography, the required fractions are collected and the solvent is evaporated.

Where stereochemistry is not stated for a spirocycle designated L1, it means a mixture of stereoisomers unless otherwise stated or clearly implied by the context.

If a stereocenter is designated RS, it means that a racemic mixture was prepared at the indicated center unless otherwise indicated.

A. Preparation of intermediate compounds.

Preparation of intermediate 1

A mixture of 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine (525 mg, 2.08 mmol), prepared as described in Journal of Medicinal Chemistry (2016), 59(3 ), 892-913, tert-butyl-2,7-diazaspiro[4.5]decane-2carboxylate (550 mg, 2.29 mmol) and DIPEA (1.43 mL, 8.3 mmol) in ACN (12 mL) were heated at 80°C overnight. The solution was cooled and the mixture was poured into chilled water, the product was extracted with EtOAc, the organic layer was dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (stationary phase: irregular particles, 15-40 μm, 50 g, mobile phase: DCM/MeOH: gradient 100/0 to 99/1). Product containing fractions were collected and evaporated to dryness to yield 770 mg (81% yield) of intermediate 1.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of intermediate 1, starting from the corresponding starting materials.

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Intermediate connection number Structure Amount (mg) Exit (%) Intermediate 2 (from CAS[336191-17-4] and [1628317-85-0]) F ₃ C S-^ _N ^ 350 100 Intermediate 3 (from CAS[885270-84-8] and [1628317-85-0]) AND ABOUT LL? 200 73 Intermediate 4 (from CAS[885270-86-0] and [1628317-85-0]) F ₃ C 660 78 Intermediate 5 (from CAS[885268-42-8] and intermediate 15) F ₃ CS^- _N Y\ 355 57

Preparation of intermediate 3

Solution of 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine (11.4 g, 44.96 mmol), tert-butyl-2,6diazaspiro[3.4]octane-2 -carboxylate (10.5 g; 49.46 mmol) and DIPEA (15.5 ml; 89.93 mmol) in iPrOH (183 ml) were heated at 90°C overnight. The solution was cooled to room temperature. and the solution was poured into water, then extracted with EtOAc (3x). The organic layer was washed with brine, dried over MgSO ₄ and filtered.

The precipitate (in the aqueous layer) was filtered off, washed with small amounts of DCM and combined with the previous filtrate. The solvent was evaporated to obtain 19.9 g of a brown solid. The residue was taken up in diethyl ether and the precipitate was filtered and dried to give 18.5 g of a pale brown solid representing intermediate 3 (96%).

Alternative preparation of intermediate 3.

To a mixture of 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine (3.00 g, 11.9 mmol) and tert-butyl-2,6diazaspiro[3.4]octane- 2-carboxylate (2.5 g, 11.8 mmol) in EtOH (50 ml) was added in one portion DIPEA (2 g, 15.5 mmol). The mixture was stirred at room temperature for 18 hours. The mixture was evaporated and the residue was dissolved in EA (200 ml). The solution was washed with water (100 mlx2), dried over Na ₂ SO ₄ , fil

- 33 045364 was pickled and evaporated to give intermediate 3 (5.10 g, 11.9 mmol, 100% yield) as a brown oil.

Preparation of intermediate 6

A mixture of intermediate 1 (770 mg, 1.69 mmol) and 4 N solution. HCl in dioxane (4.22 ml, 16.9 mmol) in ACN (45 ml) was stirred at RT. during the night. The mixture was poured into ice water, its basicity was increased with 3 N. NaOH, the product was extracted with DCM, the organic layer was dried over MgSO ₄ , evaporated to dryness to obtain 670 mg of intermediate 6, which was used in the next step without further purification.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of intermediate 6, starting from the corresponding starting materials.

Intermediate connection number Structure Quantities about (mg) Exit (%) Intermediate 7 (from intermediate 2) H 8 ^F ₃ c Salt HC1 320 Intermediate 8 (from intermediate 4) n 7 xrS F ₃ C Salt HCl 582

Preparation of intermediate 9

In a sealed tube, 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine (0.6 g, 2.37 mmol), prepared as described in Journal of Medicinal Chemistry (2016) ), 59(3), 892-913, tert-butyl-2,6diazaspiro[3.3]heptane-2-carboxylate (0.57 g, 2.85 mmol), DIPEA (0.82 ml, 4.75 mmol) in iPrOH (15 ml) was heated at 90°C for 2 hours. The solution was cooled to room temperature. and the reaction mixture was poured into water, then extracted with EtOAc. The organic layer was washed with water, dried over MgSO ₄ , filtered and evaporated to dryness. The crude product was crystallized from Et ₂ O to give 0.6 g (61% yield) of intermediate 9.

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Preparation of intermediate 10

A mixture of intermediate 9 (4.43 g, 10.69 mmol) in formic acid (24 ml) was stirred at room temperature. during the night. The reaction mixture was evaporated. The residue was taken up twice with Et ₂ O and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH; 80 g; mobile phase: 90% DCM, 10% MeOH, 1% NH4OH). The purified fractions were collected and evaporated to dryness to yield 3.34 g (99%) of intermediate 10.

Preparation of intermediate 10b

A mixture of intermediate 9 (0.55 g, 1.33 mmol) in formic acid (3 ml) was stirred at room temperature. for 20 hours. The mixture was evaporated in vacuo to leave a residue which was taken up twice with Et ₂ O and evaporated to dryness to give 0.4 g (96% yield) of intermediate 10b (formic acid salt). The crude product was used without any further purification in the next step.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of intermediate 10b, starting from the corresponding starting materials.

Intermediate connection number

Intermediate connection 12 intermediate connection 5)

Preparation of intermediate 11

A mixture of intermediate 3 (8.57 g, 20 mmol) in formic acid (51 ml) was stirred at RT. for 20 hours. The reaction mixture was stirred at room temperature. during the weekend. The mixture was evaporated and the residue was cooled to 5°C, taken up with DCM and neutralized with 3N. aqueous NaOH solution. The organic layer was washed with water, dried over MgSO ₄ , filtered and the solvent was evaporated. The residue (7.63 g orange oil) was purified by silica gel chromatography (unmodified irregular silica, 120 g, mobile phase: 1% NH4OH, 85% DCM, 15% MeOH). The purified fractions were collected and the solvent was evaporated to give 3.65 g of a yellow oil representing intermediate 11 (56%).

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Alternative preparation of intermediate 11.

TFA (17.9 ml; 233.38 mmol) was added to a solution of intermediate 3 (5 g; 11.67 mmol) in DCM (130 ml) at 5°C and the reaction mixture was stirred at RT. for 4 hours. The reaction mixture was diluted with heptane and evaporated to dryness (3x) to obtain 10.7 g of a brown oil. The residue was purified by silica gel chromatography (SiOH irregular particle, 40 μm; 220 g, mobile phase: 1% NH ₄ OH, 90% DCM, 10% MeOH). The purified fractions were collected and the solvent was evaporated. The residue (4 g) was solubilized with DCM and the product crystallized. The mixture was evaporated and absorbed several times with ACN and the solvent was evaporated to give 4 g of a pale yellow solid, intermediate 11.

Preparation of intermediate 11b

TFA (2.2 ml; 28 mmol) was added to a solution of intermediate 3 (600 mg; 1.4 mmol) in DCM (13 ml) at 0°C, then the reaction mixture was stirred at RT. during the night. The reaction mixture was evaporated to dryness to obtain 1.26 g of intermediate 11b as the TFA salt. The product was directly used without any further purification.

Preparation of intermediate 11c

A solution of 4 M HCl in dioxane (150 ml) was added to intermediate 3 (6.5 g, 15.17 mmol) at RT. The reaction mixture was stirred at room temperature. for 1 hour. The mixture was evaporated in vacuo to give 5.7 g of a yellow solid, intermediate 11c as the HCl salt. The product was used without purification for the next step.

Preparation of intermediate 13

Acetic anhydride (1 ml, 10.7 mmol) was added dropwise at room temperature. to a solution of 2-amino-5-(2,2,2trifluoroethyl)thiophene-3-carboxamide (2 g, 8.92 mmol) in toluene (50 ml) and trimethylamine (6.2 ml, 44.6 mmol). The solution was refluxed for 5 hours, poured into water, extracted with EtOAc and washed with brine (x2). The organic layer was dried over MgSO ₄ , filtered and evaporated to dryness, the crude product was taken up with EtOAc and the precipitate was filtered to give 1.5 g of intermediate 13 (63% yield/brown solid).

Preparation of intermediate 14

OH αίΛι F ₃ C

To a solution of intermediate 13 (1.5 g, 5.63 mmol) in EtOH (70 ml) at room temperature. 1 M KOH solution was added dropwise. The reaction mixture was stirred at room temperature. for 30 minutes, then the mixture was refluxed for 3 hours. The reaction mixture was cooled to room temperature, then poured into ice water, acidified with 3 N. HCl, extracted with DCM and decanted. The combined organic layers were washed with brine and dried over MgSO ₄ , filtered and evaporated to dryness. The residue was crystallized from Et ₂ O to obtain 0.7 g of prome

- 36 045364 terrible compound 14 (50% yield), which was used without additional purification in the next step.

Preparation of intermediate 15

Intermediate 14 (0.7 g, 2.82 mmol) and phosphorus trichloride (5 ml) were heated at 110°C for 2 hours. The reaction mixture was cooled to room temperature, then evaporated to dryness. The residue was taken up carefully with ice and DCM, increased in basicity with aqueous K ₂ CO ₃ (10%) and the organic layer was washed with water, dried over MgSO ₄ , filtered and evaporated to dryness to give 0.75 g (yield 99 %) of intermediate 15, which was used without further purification in the next step.

Preparation of intermediate 16

Spiro[3.3]heptan-2-ylmethylmethanesulfonate.

To a solution of spiro[3.3]heptan-2-ylmethanol (153 mg, 1.08 mmol) in 4 ml DCM was added TEA (0.464 ml, 3.2 mmol) and the reaction mixture was cooled to 0°C. Methyl sulfonyl chloride (0.184 g, 1.605 mmol) was then added dropwise and the mixture was heated to room temperature. and stirred for 2 hours. An aqueous saturated solution of NaHCO ₃ (30 ml) and DCM (30 ml) was added. The mixture was separated, the organic layer was collected, washed with brine (10 ml), dried over Na ₂ SO ₄ and evaporated to give 300 mg of intermediate 16 as a yellow oil, which was used without further purification in the next step.

Preparation of intermediate 17

Under a stream of N2, 1-[2-(acetyloxy)ethyl]-1H-n-yrrole-2-carboxaldehyde was added to a solution of intermediate 11c (400 mg) and TEA (0.38 mL, 2.76 mmol) in DCM (20 mL). (200 mg, 1.11 mmol). The mixture was stirred at room temperature. for 4 hours. NaBH(OAc) ₃ (390 mg, 1.84 mmol) was added and the mixture was stirred at room temperature. for 48 hours. It was then poured into ice water, and the mixture was separated, and the aqueous layer was extracted with DCM. The organic layers were combined, washed with brine, then dried over MgSO4 and evaporated to dryness. The residue was purified by silica gel chromatography (stationary phase: SiOH irregular particles, 15-40 µm, 24 g, mobile phase: DCM/MeOH: 97/3). The purified fractions were collected and the solvent was evaporated in vacuo to give 180 mg of intermediate 17.

Preparation of intermediate 35.

¹ H-Pyrazole-4-carbaldehyde (0.5 g; 5.2 mmol) and cesium carbonate (3.39 g; 10.4 mmol) were diluted in ACN (10 ml). 2-bromoethyl methyl ether (0.636 mL; 6.77 mmol) was then added and the reaction mixture was refluxed for 2 hours. The reaction mixture was partitioned between saturated NaHCO ₃ solution and EtOAc. The organic layer was separated, dried over MgSO ₄ , filtered and concentrated.

The residue was purified by chromatography on silica gel (SiO ₂ irregular particles, 120 g, DCM/MeOH: 100/0 to 95/5). Fractions containing the product were mixed and concentrated

- 37 045364 yielding 439 mg (55%) of intermediate 35.

Preparation of intermediate 20

Intermediate 11 (150 mg, 0.46 mmol), (+/-)-methyl-alpha-bromophenylacetate (0.08 ml, 0.50 mmol) and K ₂ CO ₃ (127 mg, 0.92 mmol) in DMF (10 ml) was stirred at RT. for 5 hours. The reaction mixture was poured into ice water and EtOAc was added. The organic layer was separated, washed with brine, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (stationary phase: SiOH irregular, 15-40 µm, 24 g, mobile phase: DCM/MeOH (+10% NH4OH): gradient 97/3 to 95/5). The purified fractions were collected and evaporated to dryness to yield 162 mg (74% yield) of intermediate 20.

Preparation of intermediate 32

Lithium hydroxide monohydrate (71 mg; 1.7 mmol) was added at room temperature. to a solution of intermediate 20 (162 mg; 0.34 mmol) in THF (3 ml) and water (3 ml). The mixture was stirred at room temperature. overnight, then concentrated and acidified with an aqueous solution of 3 N. HCl (pH 2-4). The precipitate was filtered and dried to yield 33 mg (21%) of intermediate 32 (90% purity based on LC/MS). The starting layer was evaporated to dryness to obtain 243 mg of crude fraction containing intermediate 32.

Obtaining intermediate compound 54

Intermediate 11c (333 mg), methyl 2-formyl benzoate (148.5 mg; 0.905 mmol), NaBH(OAc) ₃ (872 mg; 4.11 mmol) and trimethylamine (250 mg; 2.47 mmol) were mixed in dichloroethane (16 ml) and the reaction mixture was stirred at room temperature. during the night. Aqueous NaHCO ₃ (1 ml) was then added and the mixture was extracted with DCM (4x15 ml). The organic layers were separated, mixed, dried over MgSO ₄ , filtered and concentrated to give 450 mg of intermediate 54 as a white solid.

The intermediates listed in the table below were prepared by using a method similar to that described for the preparation of intermediate 54, starting from the appropriate starting materials.

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Intermediate number connections Structure Amount (mg) Exit (%) Intermediate 44 (from Intermediate Is and Intermediate 52) % M ss m \\ // 1 / xT 340

Preparation of intermediate 47

A mixture of intermediate 10b (150 mg), 1-(tetrahydro-2H-pyran-2-yl)-N-pyrazol-4carboxaldehyde (225 mg; 1.249 mmol) and AcOH (24 μl; 0.416 mmol) in dichloroethane (4.5 ml) was stirred at 50°C for 2 hours. The reaction mixture was cooled to room temperature and NaBH(OAc) ₃ (265 mg; 1.249 mmol) was added. The reaction mixture was stirred at room temperature overnight, poured into 10% aqueous K ₂ CO ₃ solution and extracted with DCM. The organic layer was decanted, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 24 g; mobile phase: gradient 0% MeOH, 100% DCM to 10% MeOH, 90% DCM). The purified fractions were collected and evaporated to dryness to yield 150 mg of intermediate 47.

The intermediates listed in the table below were prepared by using a method similar to that described for the preparation of intermediate 47, starting from the appropriate starting materials.

Intermediate connection number Structure Amount (mg) Exit (%) Intermediate 48 (from intermediate 10b and 1(oxan-2-yl)pyrazol-3 carbaldehyde) D ¹ o 6 158

Preparation of intermediate 25

A mixture of (4S)-1-Boc-4-methyl-L-proline (174 mg, 0.761 mmol), HBTU (288 mg, 0.761 mmol) and DIPEA (0.65 ml, 3.804 mmol) in DMF (7.5 ml ) was stirred for 1 hour. A solution of intermediate 10 (250 mg, 0.761 mmol) in DMF (5 ml) was then added. The reaction mixture was stirred at room temperature. during the night. The reaction mixture was poured into ice water, increased in basicity with 10% aqueous K ₂ CO ₃ and extracted with EtOAc. The organic layer was washed with water,

- 39 045364 with that saline solution, dried over MgSO ₄ , filtered and evaporated to dryness. The residue (490 mg) was purified by silica gel chromatography (SiOH irregular particle, 40 g; mobile phase: NH ₄ OH/DCM/MeOH: 0.5/95/5). The purified fractions were collected and evaporated to dryness to yield 330 mg (82% yield) of intermediate 25.

The intermediates listed in the table below were prepared by using a method similar to that described for the preparation of intermediate 25, starting from the corresponding starting materials.

Quantity

Obtaining intermediate compound 104

Connection number

Intermediate connection of intermediate connection 10b and

2S,4S)-4fluoropyrrolidine-2carboxylic acid)

Intermediate connection intermediate connection 11

Vos-azaspiro| 2.41 heptane 6-carboxylic acid)

DIPEA (0.48 ml; 2.775 mmol) was added to a solution of intermediate 10b (200 mg), 3-carboxybenzaldehyde (100 mg; 0.666 mmol) and HATU (317 mg; 0.833 mmol) in DMF (10 ml) and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was poured into water and extracted with EtOAc. The organic layer was decanted, washed with water, then with brine, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 24 g; mobile phase: gradient 0% MeOH, 100% DCM to 10% MeOH, 90% DCM). The purified fractions were collected and evaporated to dryness to yield 62 mg of intermediate 104.

Preparation of intermediate 41

In flow N2 at r.t. to a solution of intermediate 11 (250 mg, 0.76 mmol) in DCM (12 ml) was added intermediate 42 (246 mg, 0.91 mmol). The mixture was stirred at room

- 40 045364 temperature for 3 hours. The mixture was cooled to 5°C, NaBH(OAc) ₃ (323 mg, 1.52 mmol) was added and the mixture was stirred at room temperature. during the night. Then it was poured into ice water and the layers were separated. The aqueous layer was extracted with DCM. The organic layers were combined, washed with brine, then dried over MgSO ₄ and evaporated. The residue was crystallized from Et ₂ O and pentane. The white precipitate was filtered off and dried under vacuum to give 55 mg (100% yield) of intermediate 41.

Preparation of intermediate 43

Intermediate 11 (500 mg, 1.52 mmol), 1H-pyrrole-1-carboxylic acid 2-(chloromethyl)-1,1-dimethylethyl ester (493 mg, 2.28 mmol) and K2CO3 (1.05 g , 7.61 mmol) in ACN (12 ml) was stirred at room temperature for 24 hours. The reaction mixture was poured into ice water and EtOAc was added. The organic layer was separated, washed with brine, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (stationary phase: SiOH irregular particle, 15-40 µm, 24 g, mobile phase: NH ₄ OH/DCM/MeOH: gradient 0.1/97/3 to 0.1/95 /5). The purified fractions were mixed and evaporated to give 100 mg (14% yield) of intermediate 43.

The intermediates listed in the table below were prepared by using a method similar to that described for the preparation of intermediate 43, starting from the appropriate starting materials.

- 41 045364

Intermediate number connections Structure Amount (mg) Exit (%) Intermediate 51 (from 1H-pyrrole-4- carboxaldehyde and 3-bromopropionitrile) 157 66 At 60°C as reaction temperature 1000 71 At Intermediate 42 (from 1H-pyrrole-4- carboxaldehyde and N-(2bromoethyl)phthalimide) temperature of formation of phlegm as reaction temperatures Intermediate 52 (out of 3- hydroxybenzaldehyde and 3(Boc-amino)propyl bromide) 340 99 At 75°C as reaction temperature 1563 65 Intermediate 53 (from 1H-pyrazole-4- carbaldehyde and 2- bromoethoxy-tert-butyldimethylsilane) At reflux temperature as reaction temperature

Preparation of intermediate 50

Under a stream of N2, tert-butyl-4-formyl-1H-pyrazole-1-carboxylate (133 mg, 0.68 mmol) and AcOH were added to a solution of intermediate 11 (202 mg, 0.62 mmol) in DCM (10 mL). (35 µl, 0.62 mmol). The mixture was stirred at room temperature for 2 hours. NaBH(OAc)3 (521 mg, 2.46 mmol) was added and the mixture was stirred at room temperature. overnight, poured into ice water, and the layers were separated. The aqueous layer was extracted with DCM. The organic layers were combined, washed with brine, then dried over MgSO ₄ and evaporated. The residue was purified by chromatography on silica gel (stationary phase: SiOH irregular particles, 15-40 µm, 24 g, mobile phase: DCM/MeOH (+10% NH4OH): 97/3). The purified fractions were mixed and evaporated to give 145 mg (46% yield) of intermediate 50.

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Preparation of intermediate 55

In a sealed tube under N2 flow, intermediate 53 (349 mg, 1.37 mmol) and Ti(OiPr) ₄ (436 μL, 1.83 mmol) were added to a solution of intermediate 11 (300 mg, 0.914 mmol) in THF ( 6 ml). The solution was stirred at 50°C for 5 hours, then at room temperature. during the night.

The reaction mixture was cooled to 5°C and 2 N was added dropwise. iPrMgCl in THF (2.28 ml, 4.57 mmol). The reaction mixture was slowly heated to room temperature. and it was stirred overnight. The reaction mixture was diluted with EtOAc and poured into a 10% aqueous K ₂ CO ₃ solution. Insoluble matter was removed by filtration through Celite®. The organic layer was decanted, washed with brine, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 40 g; mobile phase: MeOH/DCM: gradient 0/100 to 10/90). The purified fractions were collected and evaporated to dryness to yield 0.3 g (54% yield) of intermediate 55.

Preparation of intermediate 63 and intermediate

intermediate connection 63 a

A solution of tert-butyl-3-acetylasetidine-1-carboxylate (364 mg; 1.83 mmol), intermediate 11 (400 mg; 1.22 mmol), titanium(IV) isopropoxide (725 μL; 2.44 mmol) in ethanol (2 ml) was stirred at 45°C for 30 min (the solution turned dark yellow). Ethanol (12 ml) and NaBH4 (138 mg; 3.66 mmol) were added and the solution turned pale yellow. The reaction mixture was stirred at room temperature overnight. It was then poured into a 10% aqueous solution of K ₂ CO ₃ and DCM.

- 43 045364

The insoluble material was filtered through a Celite® pad. The organic layer was decanted, filtered through Chromabond® and the solvent was evaporated to 624 mg of a pale yellow oil, which was purified by chromatography on silica gel (SiO ₂ ; 25 g; mobile phase: gradient from 98% DCM, 2% MeOH to 96% DCM, 4% MeOH). Fractions containing product were collected and the solvent was evaporated to give 223 mg (36%) of intermediate 63 as a white foam.

Intermediate 63 was purified using chiral SFC (stationary phase: CHIRALCEL OJ-H, 5 µm, 250x20 mm, mobile phase: 92% CO ₂ , 8% MeOH (0.3% iPrNH ₂ )). The purified fractions were collected and the solvent was evaporated to give 83 mg (13%) of intermediate 63a as a colorless oil and 90 mg (14%) of intermediate 63b.

The intermediates listed in the table below were prepared by using a method similar to that described for the preparation of intermediate 63, starting from the corresponding starting materials. ________________________________________________

Intermediate connection number Structure Quantity Exit Intermediate 60 (from intermediate 11 and N-Boc-4acetylpiperidine) j I T ω 1 600 mg 52% Intermediate 60a and Intermediate 60b from Chiral SFC Resolution intermediate Intermediate connection 60a 221 mg 20% compounds 60: (Chiralpak AD-H 5 µm 250*30 mm; mobile phase: 75% CO ₂ , 25% iPrOH (0.3% iPrNH ₂ )). Intermediate 109 (from Intermediate 11 and Intermediate 108) F ₃ C S-^ _N ^ Intermediate 60b ;--------- λ—fe '—< o—4— ъ ^S — 229 mg 324 mg 20% 33%

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Getting intermediate 68 intermediate

intermediate 68a and compound 61

A solution of intermediate 113 (1.67 g, 6.54 mmol) in THF (15 mL) was added to a solution of intermediate 11 (1.4 g, 4.36 mmol) and TFA (2 mL, 26.16 mmol) in THF (30 ml). The reaction mixture was stirred at room temperature. during the night. NaBH(OAc) ₃ (2.77 g, 13.08 mmol) was then added portionwise. The reaction mixture was stirred at room temperature. within 10 days. The solution was poured into a 10% aqueous K ₂ CO ₃ solution and EtOAc was added. The mixture was extracted with EtOAc (3x). The organic layers were combined, washed with brine, dried over MgSO ₄ , filtered and the solvent was evaporated. The residue (2.9 g; yellow oil) was purified by chromatography on silica gel (SiO ₂ ; 40 g; eluent: 97% DCM, 3% MeOH, 0.3% NH4OH to 90% DCM, 10% MeOH, 1% NH4OH ). The required fractions were collected and the solvent was evaporated to obtain 266 mg of a colorless oil representing Intermediate 68 and 215 mg of a colorless oil representing Fraction 1.

Intermediate 68 was purified using chiral SFC (Lux-cellulose-25 µm, 250x30 mm, mobile phase: 50% CO2, 50% MeOH (0.3% iPrNH ₂ )). The purified fractions were collected and the solvent was evaporated to provide 114 mg (5%) of a colorless oil representing intermediate 68a and 109 mg (4%) of a colorless oil representing intermediate 68b.

Fraction 1 was purified using reverse phase (YMC-actus Triart C18 10 µm, 30x150 mm, mobile phase: gradient from 65% NH4HCO3 0.2%, 35% ACN to 25% NH4HCO3 0.2%, 75% ACN). Fractions containing the product were collected and the solvent was evaporated. Residue (160 mg; colorless oil) Lyofi

- 45 045364 was lysed with water-ACN to give 90 mg (6%) of a white solid as compound 61.

Preparation of intermediate 69 intermediate and intermediate

A solution of 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine (5.07 g, 20.08 mmol), 2-BOC-2,7diazaspiro[4.4]nonane (5 g; 22.09 mmol) and DIPEA (6.9 ml; 40.17 mmol) in iPrOH (80 ml) were heated at 90°C overnight. The solution was cooled to room temperature. and the solution was poured into water, then extracted with EtOAc (3x). The organic layer was washed with brine, dried over MgSO ₄ and the solvent was evaporated to dryness. The residue (9 g, pale brown solid) was taken up in diethyl ether and the precipitate was filtered and dried to give 8.4 g of intermediate 69 (95%, off-white solid).

Intermediate 69 was purified using chiral SFC (Chiralpak IG 5 µm, 250x20 mm, mobile phase: 65% CO ₂ , 35% iPrOH (0.3% iPrNH ₂ )). The purified fractions were collected and the solvent was evaporated to yield 4.07 g of intermediate 69a (46%, yellow foam) and 4.29 g of intermediate 69b (48%, yellow foam).

Preparation of intermediate 70a

Intermediate 70a was prepared by using a method similar to that described for the alternative preparation of intermediate 11, starting from the corresponding starting material of intermediate 69a.

The intermediate shown in the table below was prepared by using a method similar to that described for the alternative preparation of intermediate 11, starting from the appropriate starting materials.

- 46 045364

Intermediate connection number Structure Quantity Exit d Intermediate 70b (from intermediate 69b) / \ ³ >70 F ₃ C 3.4 g Kolic

Preparation of Intermediate 71 Intermediate 71a and Intermediate

Method A.

A mixture of intermediate 11b (4.2 g) and intermediate 72 (1.6 g, 7 mmol) in THF (50 ml) was stirred at RT. during the night. NaBH(OAc) ₃ (3 g; 14 mmol) was then added in portions. The reaction mixture was stirred at room temperature for 24 hours. The solution was poured into cold water, its basicity was increased with 3 N. aqueous NaOH solution and EtOAc was added. The organic layer was separated, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 80 g, mobile phase: gradient 98% DCM, 2% MeOH (+10% NH4OH) to 95% DCM, 5% MeOH (+10% NH4OH)). Purified fractions were collected and evaporated to dryness to yield 852 mg of intermediate 71. Enantiomers were separated using chiral SFC (CHIRALCEL OD-H 5 µm, 250x30 mm; mobile phase: 70% CO2, 30% EtOH). The purified fractions were collected and evaporated to dryness to obtain 294 mg of intermediate 71a and 303 mg of intermediate 71b.

- 47 045364

Method B.

The experiment was performed 6 times with the same amount (640 mg; 1.95 mmol). Ti(OEt) ₄ (0.8 ml; 3.9 mmol) was added at room temperature to a solution of intermediate 11 (640 mg; 1.95 mmol) and intermediate 72 (665 mg; 2.92 mmol) in DCE ( 20 ml) and MeOH (8 ml). The reaction mixture was stirred at room temperature. for 24 hours, cooled to 10°C, then NaBH ₃ CN (367 mg; 5.84 mmol) was added in portions. The reaction mixture was stirred at room temperature for 8 days. The solutions were collected for processing: poured into cold water, increased in basicity with K ₂ CO ₃ powder and extracted with DCM. The suspension was filtered through a Celite® pad. The filtrate was decanted, dried over MgSO ₄ , filtered and evaporated to dryness.

The residue was purified by silica gel chromatography (irregular SiOH, 40 g; mobile phase: gradient 100% DCM, 0% MeOH to 97% DCM, 3% MeOH, 0.1% NH4OH). The purified fractions were collected and evaporated to dryness to yield 1.7 g (28%) of intermediate 71.

Enantiomers were separated using chiral SFC (Chiralcel OD-H 5 µm, 250x30 mm; mobile phase: 70% CO2, 30% EtOH (0.3% iPrNH2)). The purified fractions were collected and evaporated to dryness to yield 697 mg (11%) of intermediate 71a and 727 mg (12%) of intermediate 71b.

The intermediates shown in the table below were prepared according to Method A described for the preparation of intermediates 71, 71a and 71b, starting from the corresponding starting materials.

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intermediate connection 89 Intermediate 82, BOS \ /^ ^F )—\ ^RS From intermediate 11b and intermediate 83 BOC 1 N 40 mg intermediate \R*/ ^F connection 82a And ъ Υ/ώ F ₃ C S-^ _N ^ BOS γ intermediate connection 82b ъ From separation via chiral SFC intermediate 82 (stationary phase: CHIRALPAK ADH 5 µm, 250*30 mm, mobile phase: 70% CO2, 30% iPOH (0.3% iPrNH ₂ ))

Preparation of intermediate 77

- 49 045364 intermediate connection 77 a and intermediate connection 77b

Reaction mixture 1. In a sealed tube, a solution of intermediate 78 (2 equiv.), intermediate 11 (100 mg, 0.305 mmol) and Ti(OiPr) ₄ (6 equivalents) in EtOH (0.2 ml) was heated at 45° C for 1 hour. The mixture was cooled to room temperature, diluted with EtOH (3 ml) and NaBH ₄ (2 eq.) added. The reaction mixture was stirred at room temperature for 4 hours, yielding 60% of intermediate 77 according to LC/MS results.

Reaction mixtures 2 and 3. The reaction was carried out twice with the same amount. In a sealed tube, a solution of intermediate 78 (2 equiv.), intermediate coco11 (450 mg; 1.37 mmol) and Ti(OiPr) ₄ (6 equivalents) in EtOH (0.9 ml) was heated at 45 °C in within 1 hour. The mixture was cooled to room temperature, diluted with EtOH (13 ml) and NaBH4 (2 eq) added. The reaction mixture was stirred at room temperature for 18 hours.

The three reaction mixtures were diluted with EtOAc and poured into a mixture of 10% K ₂ CO ₃ and saline. The suspension was sonicated for 30 min and filtered through a Celite® pad. The organic layer was decanted, washed with 10% aqueous K ₂ CO ₃ then brine, dried over MgSO ₄ , filtered and evaporated to dryness. The residue (2.6 g) was purified by silica gel chromatography (SiOH irregular particle, 50 g; mobile phase: gradient 0% NH4OH, 0% MeOH, 100% DCM to 0.7% NH4OH, 7% MeOH, 93 % DCM).

The purified fractions were collected and evaporated to dryness. The residue (1.5 g; 89%) was purified a second time by silica gel chromatography (irregular SiOH, 40 g; mobile phase: 60% heptane, 35% EtOAc, 5% MeOH (+10% NH4OH)). Fractions containing product were collected and evaporated to dryness to yield 980 mg (58%; 82% purity based on LC/MS) of intermediate 77.

The crude fraction containing intermediate 77 was further purified using achiral SFC (diethylaminopropyl, 5 µm, 150x30 mm; mobile phase: 90% CO2, 10% MeOH). The purified fractions were collected and evaporated to dryness to yield 620 mg (37%) of intermediate 77.

Enantiomers were separated using chiral SFC (Lux Cellulose-25 µm, 250x30 mm; mobile phase: 50% CO2, 50% MeOH (0.3% iPrNH2)). Fractions containing the products were collected and evaporated to dryness to yield 276 mg (16%) of intermediate 77a and 269 mg (16%) of intermediate 77b.

The intermediates listed in the table below were prepared by using a method similar to that described for the preparation of intermediates 77, 77a and 77b, starting from the corresponding starting materials.

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Intermediate connection number Structure Quantity Exit Intermediate connection 80 BOC 4 ъ F ₃ CS^ _N ·^ From intermediate 11 and tert-butyl-3-propanoyl azetidin-1 carboxylate 240 mg 57%

Preparation of intermediate compound 72

Under N ₂ atmosphere at 5°C, 2 M iPrMgCl in THF (19 ml; 38.33 mmol) was added to a solution of intermediate 73 (4.6 g; 18.83 mmol) in THF (70 ml). The solution was stirred at 5°C for 30 min, its temperature was allowed to slowly rise to room temperature, stirred for 1 hour, then heated at 40°C for 5 hours.

The reaction mixture was cooled to room temperature, poured into a mixture of ice water and saturated aqueous NH ₄ Cl and extracted with EtOAc. The organic layer was decanted, dried over MgSO ₄ , filtered and evaporated to dryness to give 4.7 g of intermediate 72 (quantitative).

The intermediates listed in the table below were prepared by using a method similar to that described for the preparation of intermediate 72, starting from the appropriate starting materials.

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Intermediate connection number Structure Quantity Exit Intermediate connection 75 O \ O M O-- From intermediate 73 and 0.5 cyclopropylmagnesium bromide in THF 25 g Quantity Intermediate connection 78 Ad ^N -1 ° / From intermediate 73 and 0.4 M isobutylmagnesium bromide 19 g 96% Intermediate connection 83 —( 'ϊ θ o ⁷ O-- From intermediate 84 and 2 M isopropylmagnesium chloride in THF 137 mg 49% Intermediate connection 105 \ /^° /SY From A-methoxy-N- methyltetrahydrofuran-3 carboxamide and 2 M isopropylmagnesium chloride in THF 71 mg 16%

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Intermediate connection 108 ABOUT From A-methyl-X-methoxy-1-(tert- butoxycarbonyl)piperidine-4acetamide and 2 M isopropylmagnesium chloride in THF 710 mg 26% Intermediate connection 111 Ol o ⁰ \ From intermediate 73 and chloro[(4-methylphenyl)methyl]magnesium 770 mg 40% Intermediate connection FROM ----( 7\O V--\ ^N --/ o ⁷ '— ⁷ O—4— From 1,1-dimethylethyl ester 4-[(A-methoxy-A-methylamino)carbonyl]-1 piperidinecarboxylic acid acid and 2 M isopropylmagnesium chloride in THF 1230 mg 33%

Preparation of intermediate compound 73

1-Boc-azetidine-3-carboxylic acid (5 g; 24.9 mmol) and N,O-dimethylhydroxylamine hydrochloride (3.64 g; 37.3 mmol) were placed in a round bottom flask under N2 atmosphere. DCM (75 mL) was added followed by EDCI.HCl (7.15 g, 37.3 mmol), DMAP (155 mg, 1.27 mmol) and DIPEA (6.5 mL, 37.4 mmol). The reaction mixture was stirred at room temperature. for 16 h and diluted with DCM (100 ml). The organic layer was washed with aqueous 1 M HCl solution (2x50 ml), sat. NaHCO ₃ solution (50 ml) and saline solution (50 ml). The organic phase was decanted, dried over MgSO ₄ , filtered and evaporated to dryness to give 6.04 g (99%) of intermediate 73.

The intermediates listed in the table below were prepared by using a method similar to that described for the preparation of intermediate 73, starting from the corresponding starting materials.

- 53 045364

Under N2 atmosphere, a solution of intermediate 11 (204 mg; 0.62 mmol), intermediate 86 (217 mg; 0.81 mmol) and Ti(OEt) ₄ (0.26 mL; 1.24 mmol) in DCE (7 ml) was stirred at room temperature. during the night. NaBH ₃ CN (129 mg; 2 mmol) was added and the solution was stirred for 4 days. Water was added dropwise, then the solution was filtered through a Celite® pad. The filtrate was separated. The organic layer was washed with water, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 24 g; mobile phase: gradient 97% DCM, 3% MeOH (+10% NH4OH) to 95% DCM, 5% MeOH (+10% _NH4OH ) ). The purified fractions were collected and evaporated to dryness to yield 209 mg (80%) of intermediate 85.

The intermediate shown in the table below was prepared by using a method similar to that described for the preparation of intermediate 85, starting from the appropriate starting materials.

From intermediate 11 and 1,1-dimethylethyl 4[(1-methyl-1H-pyrazol-4-yl)carbonyl]-1 piperidinecarboxylic acid ester

Intermediate connection number Intermediate connection 87

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Preparation of intermediate 86

2-Bromo-1-(1-methyl-III-pyrazol-4-yl)ethanone (0.5 g; 2.46 mmol) in DMF (10 ml) was added to potassium phthalimide (0.46 g; 2.46 mmol). The reaction mixture was stirred at room temperature. for 5 hours, poured into ice water and added EtOAc. The organic layer was separated, washed with water, brine, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (SiOH irregular particle, 15-40 µm, 24 g; mobile phase: 97% DCM, 3% MeOH (+10% NH ₄ OH)). The purified fractions were collected and evaporated to dryness to yield 460 mg (69%) of intermediate 86.

Preparation of intermediate 89

Under an N2 atmosphere at -70°C, 1.6 M n-BuLi in hexane (6 .2 ml; 9.92 mmol). The reaction mixture was stirred at -70°C for 1 hour, then a solution of intermediate 73 (2 g, 8.19 mmol) in THF (10 ml) was added dropwise. The reaction mixture was stirred at -70°C for 2 hours, allowed to warm to room temperature and stirred overnight. The solution was poured into a mixture of ice water and saturated NH ₄ Cl solution, then EtOAc was added. The organic layer was decanted, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 50 g; mobile phase: gradient 100% DCM, 0% MeOH to 98% DCM, 2% MeOH, 0.1% NH4OH). The purified fractions were collected and evaporated to dryness to yield 320 mg (15%) of intermediate 89.

Preparation of intermediate 92

Under N2 atmosphere, HBTU (210 mg; 0.555 mmol) was added to a solution of BOC-L-proline (119 mg; 0.555 mmol) and DIPEA (0.48 mL; 2.775 mmol) in DMF (10 mL). The solution was stirred for 30 min, then intermediate 10b (200 mg) was added and the solution was stirred at room temperature over the weekend. The reaction mixture was then poured into ice water, rebasified with 10% aqueous K2CO3 and extracted with EtOAc. The organic layer was washed with water, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 24 g; mobile phase: gradient 0.5% NH4OH, 5% MeOH, 95% DCM to 1% NH4OH, 10% MeOH, 90% DCM). The purified fractions were collected and evaporated to dryness to yield 168 mg of intermediate 92.

The intermediates listed in the table below were prepared by using a method similar to that described for the preparation of intermediate 92, starting from the corresponding starting materials.

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Intermediate connection number Structure Quantity Exit Intermediate 93 A? and ^s \,1 voe Li F ₃ CS From intermediate 10b and (28.4K)-M-boc-4-methylpyrrolidine-2carboxylic acid 510 mg Intermediate connection 101 ύ П ^s \rll BOC AO F ₃ C S-^ _N ^ From intermediate 10b and (2S,4R)-1-(tert-butoxycarbonyl)-4fluoropyrrolidine-2-carboxylic acid 150 mg Intermediate 96 A\ ^S ^T s\ i voe F ₃ ^CS ' From intermediate 10 and cis-1-M-Boc-4-methoxy-b-proline 283 mg 94% Intermediate connection 102 I 1 voc oVj F ₃ C From intermediate 10b and (8)-5-Boc-5-azaspiro[2.4]heptane-6carboxylic acid 200 mg

-56045364

Intermediate 103 ⁰ g? voc L / Q zhao F ₃ C From intermediate 11 c and (8)-5-(tert-butoxycarbonyl)-5azaspiro[2.4]heptane-6-carboxylic acid 160 mg Intermediate 98 o ' J BOC аСО F ₃ C Β-ΆμΑ From intermediate 10b and (K)-5-(tert-butoxycarbonyl)-5azaspiro[2.4]heptane-6-carboxylic acid 348 mg Intermediate connection 99 o L Ί ',1 vo F ₃ C From intermediate 10 and (2S,4R)-1 -(tert-butoxycarbonyl)-4methoxypyrrolidine-2-carboxylic 389 mg 75%

- 57 045364

Preparation of intermediate 107

Under a nitrogen atmosphere, a solution of intermediate 11 (250 mg; 0.761 mmol), N-Boc-4 formylpiperidine (195 mg; 0.914 mmol) in THF (7 ml) was stirred at RT. for 3 hours. NaBH(OAc) ₃ (323 mg; 1.52 mmol) was added and the mixture was stirred at room temperature. during the night. A 10% aqueous solution of K2CO3 and DCM was added. The organic layer was separated, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 24 g; mobile phase: gradient 100% DCM 0% MeOH (0% NH ₄ OH) to 90% DCM 10% MeOH (10% NH ₄ OH)). Fractions containing the product were mixed and evaporated to dryness to yield 383 mg (96%) of intermediate 107.

Preparation of intermediate 110

Add DIPEA (0.45 mL, 3.24 mmol) to an ice-cold solution of 2-(4-fluorophenyl)propanol (CAS[59667-20-8]) (0.25 g, 1.62 mmol) in DCM (1 .4 ml) followed by the addition of methanesulfonyl chloride (0.155 ml, 1.95 mmol). The mixture was stirred overnight at room temperature. The mixture was diluted with DCM (20 ml) and washed with saturated sodium bicarbonate solution (15 ml). The solution was dried over MgSO ₄ , filtered and concentrated under reduced pressure to give 0.377 g of intermediate 110. This product was used without further purification in the next step.

- 58 045364

Obtaining intermediate 112

A solution of intermediate 111 (607 mg, 2.1 mmol) and titanium isopropoxide (1.25 mL, 1.37 mmol) in EtOH (4.6 mL) was added dropwise at room temperature (over a period of 5 to 10 min ) to a mixture of intermediate 11 (459 mg, 1.4 mmol) and NaBH ₃ CN (264 mg, 4.2 mmol) in EtOH (9.2 ml). The mixture was stirred for 1 hour at room temperature. The reaction mixture was diluted with

DCM and poured into a 10% aqueous K ₂ CO ₃ solution. The suspension was filtered through a Celite® pad. The organic layer was decanted, washed with brine, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 24 g; mobile phase: DCM/MeOH: gradient 100/0 to 90/10). The purified fractions were collected and evaporated to dryness to yield 0.521 g (62%) of intermediate 112 (62%).

B. Obtaining connections.

Example B1.

Receiving connection 1

TEA (88.5 mg, 0.875 mmol) and benzaldehyde (46.4 mg, 0.44 mmol) were added sequentially to a solution of intermediate 11 (190 mg, 0.44 mmol) in anhydrous DCM (4 mL) and the mixture was stirred at k.t. within 30 min. NaBH(OAc) ₃ (185.4 mg, 0.875 mmol) was then added and the mixture was stirred at RT. during the night. Added sat. aq. a solution of NaHCO ₃ (10 ml) and DCM (10 ml) and the mixture was decanted. The aqueous layer was extracted twice with DCM (10 ml). The organic layers were combined, washed with brine (10 ml), dried over Na ₂ SO ₄ and evaporated to give a yellow oil. The crude residue was purified by silica gel chromatography (Gemini column 150x25, 5 μm, mobile phase: water (0.05% ammonium hydroxide, o6./o6.)/ACN: gradient 55/45 to 25/75). The residue was then lyophilized to give 65 mg of compound 1 (35% yield) as a yellow solid.

Example B2.

Receiving Connection 2

Intermediate 11 (100 mg, 0.3 mmol), 3,3,3-trifluoropropanal (51 mg, 0.46 mmol) in dry DCM (3 mL) was stirred at RT. for 1 hour, then NaBH(OAc) ₃ (129 mg, 0.61 mmol) was added and the mixture was stirred at room temperature. during the night. The mixture was poured into water, then extracted

- 59 045364 using DCM, the organic layer was dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (stationary phase: SiOH irregular particles, 15-40 μm, 24 g, mobile phase: NH ₄ OH/DCM/MeOH: 0.1/97/3). Product containing fractions were collected and evaporated to dryness to yield 58 mg (45%) of Compound 2, which was lyophilized with 20/80 ACN/water to yield 45 mg of Compound 2.

Receiving connection 13

A mixture of intermediate 11c (600 mg), isobutyraldehyde (160 mg; 2.221 mmol),

NaBH(OAc) ₃ (1.57 g; 7.405 mmol) and Et ₃ N (0.64 ml; 4.443 mmol) in DCE (12 ml) were stirred at room temperature overnight. A saturated aqueous solution of NaHCO ₃ (20 ml) and DCM (20 ml) was added. The organic layer was decanted and the aqueous layer was extracted with DCM (20 mlx2). The combined organic layers were washed with brine (30 ml), dried over Na ₂ SO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (mobile phase: gradient petroleum ether/EtOAc 100/0 to 0/100, then EtOAc/MeOH 100/0 to 85/15). Purified fractions were collected, evaporated to dryness and lyophilized to yield 320 mg of compound 13.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 2, starting from the corresponding starting materials.

Connection number Structure Quantities about (mg) Exit (%) Compound 3 (from intermediate 7) 1 11 F ₃ C S-^|sK in the form of hydrochloride salt 80 Compound 4 (from intermediate 11) o ω % \= _ζ 1 126 47

- 60 045364

Compound 5 (from intermediate 11) o AYj F ₃ C 75 43 Compound 6 (from intermediate 11) Z^N p N—_ yC0 F ₃ C 55 26 Compound 7 (from intermediate 11) ex r N—-. Aj0 F ₃ C 55 31

- 61 045364

Connection 8 (from Cl N—< O\=N 75 43 intermediate compound I) N- ⁴ N F ₃ C s- € N 7 75 39 Connection 9 of intermediate connection 11) (from G N- =N 'N F ₃ C s-J N— c % 40 29 Connection 10 of intermediate connection 11) (from <N- ⁴ N ^Z 'N F ₃ C sJ

- 62 045364

Compound 11 (from intermediate 11) N—x // y-ci \==N N—_ F ₃ C 65 37 Compound 12 (from intermediate 11) Ν—x o V=N Ν—_ F ₃ C 80 41 Compound 13 (from intermediate AND) F ₃ C 80 54

- 63 045364

70 18 Compound 23 (from intermediate 11 and 1methyl-1H-imide azole5-carboxaldehyde) ъ ϊ f ₃ c \ji J Compound 35 (from intermediate 11 and 1methyl-1H-pyrazole-3 carbaldehyde) \y g -ZXl i 155 40 F ₃ C \JI J Compound 69 (from intermediate 11 and phenylacetaldehyde (CAS[122-78-l])) F ₃ C . -Ό0 60 22

Example B3.

Receiving connection 14

Intermediate 10b (0.42 g), benzyl bromide (0.19 ml, 1.6 mmol) and K ₂ CO ₃ (0.55 g, 4.0 mmol) in ACN (20 ml) were stirred at RT. during the night. The mixture was poured into water, extracted with EtOAc, the organic layer was washed with brine, then dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (stationary phase: SiOH irregular particles, 15-40 μm, 40 g, mobile phase: NH ₄ OH/DCM/MeOH: 0.1/97/3). Product containing fractions were collected and evaporated to dryness to yield 170 mg (31%) of compound 14, which was crystallized from DIPE, filtered and dried to yield 103 mg of compound 14.

The compounds and intermediates listed in the table below were prepared by using a method similar to that described for the preparation of compound 14, starting from the corresponding starting materials.

- 64 045364

Connection number Structure Amount (mg) Exit (%) Connection 15 (from intermediate connection 11) r- ^N (Г y-ci \==N N—_ F ₃ C S-^ _N > 65 37 Connection 16 (from intermediate connection 8) N F ₃ C in the form of hydrochloride salt (1.7 HC1. 1.9 H ₂ O) 57 Compound 17 (from intermediate 11) (ind. 11 was reacted with 1628318-10-4 s subsequent splitting with using TFA) N III N-_ F ₃ C S-^ _N ^ 60 32

- 65 045364

Compound 18 (from intermediate 6) lch) π ^Ν '—[ Rsfxz F ₃ C 130 49 Compound 19 (from intermediate 12) L« 45 12 Compound 30 (from intermediate 11 and chloromethyltrimethylgermane) LL? 28 13 Compound 31 (from intermediate 11 and chloromethyltrimethylgermane) w ¹ O Ύ / n 36 21

Example B4.

Receiving connection 20

A mixture of 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-th]pyrimidine (150 mg, 0.59 mmol), prepared as described in Journal of Medicinal Chemistry (2016), 59(3 ), 892-913, 2-benzyl-2,7-diazaspiro-[4.4]nonane (CAS[885275-27-4]) (129 mg, 0.59 mmol) and DIPEA (0.31 ml, 1.78 mmol) in ACN (15 ml) was heated at 80°C overnight. The mixture was cooled and poured into chilled water, the product was extracted with EtOAc, the organic layer was dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (stationary phase: unmodified irregular silica, 24 g, mobile phase: DCM/MeOH/NH ₄ OH: 97/3/0.1). Product containing fractions were collected and evaporated to dryness to give 200 mg (78% yield) of compound 20 (racemic mixture).

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Preparation of enantiomers: 20a.

and 20b

Enantiomers were separated by chiral SFC (stationary phase: Lux Cellulose-45 µM,

250x21.2 mm, mobile phase: CO ₂ /MeOH (0.3% iPrNH ₂ ): 70/30). Product containing fractions were collected and evaporated to dryness to obtain 80 mg (31% yield) of the first eluted fraction F1 and 81 mg (31% yield) of the second eluted fraction F2.

F1 (80 mg; 0.185 mmol) was dissolved in acetone at 10°C and 4 N was added. HCl in dioxane (2 eq., 0.37 mmol, 93 µl) followed by EtOAc. The mixture was evaporated to dryness and taken up with Et ₂ O, the precipitate was filtered and dried to give 65 mg (20% yield) of compound 20a as the hydrochloride salt (1.95 HCl, 1.25 H ₂ O, 0.19 dioxane, 0.06 Et ₂ O).

F2 (81 mg, 0.187 mmol) was dissolved in acetone at 10°C and 4 N was added. HCl in dioxane (2 eq., 0.37 mmol, 93 µl) followed by EtOAc. The mixture was evaporated to dryness, taken up with EtOAc, the precipitate was filtered and dried to give 49 mg (15% yield) of 20b as the hydrochloride salt (2.0 HCl. 1.8 H ₂ O).

Example B5.

Receiving connection 18

A mixture of intermediate 6 (222 mg, 0.63 mmol), benzyl bromide (82 µl, 0.685 mmol) and K2CO3 (430 mg, 3.11 mmol) in ACN (20 ml) was stirred at room temperature. during the night. The solution was poured into cool water, the product was extracted with EtOAc, the organic layer was dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (stationary phase: irregular particles 15-40 μm, 30 g, mobile phase: DCM/MeOH/NH ₄ OH: gradient 100/0/0 to 97/3/0.1). Product containing fractions were collected and evaporated to dryness to give 170 mg (61% yield) of compound 18 (racemic mixture).

- 67 045364

Preparation of enantiomers representing compound 21a

and connection 21b

Compound 18 was resolved into its enantiomers by chiral SFC (stationary phase: Lux

Cellulose-25 µm, 250x30 mm, mobile phase: CO ₂ /MeOH: 75/25). Product-containing fractions were collected and evaporated to dryness to obtain 72 mg (26% yield) of the first eluted fraction F1 and 76 mg (27% yield) of the second eluted fraction F2.

F1 was dissolved in acetone (3 ml), a 4 N solution was added dropwise at 10°C. HCl in dioxane (2 equiv., 80 μl, 0.32 mmol), Et ₂ O was added and after 30 min the precipitate was filtered and dried to obtain 54 mg (yield 16%) of compound 21a as the hydrochloride salt (1.8 HCl. 1.9 H ₂ O).

F2 was dissolved in acetone (3 ml), a 4 N solution was added dropwise at 10°C. HCl in dioxane (2 equiv., 85 μl, 0.34 mmol), Et ₂ O was added and after 30 min the precipitate was filtered and dried to obtain 34 mg (10% yield) of compound 21b as the hydrochloride salt (1.8 HCl. 2.1 H ₂ O).

Example B6.

Receiving connection 22

To a solution of intermediate 11 (200 mg, 0.51 mmol) in ACN (5 ml) was added intermediate 16 (220 mg, 1.08 mmol) and K2CO3 (221.4 mg, 1.53 mmol). The mixture was heated to 90°C and stirred overnight. Water (10 ml) and DCM (10 ml) were added to the reaction mixture. The organic phase was separated and the aqueous layer was extracted with DCM (10 ml). The organic layers were combined, washed with brine (10 ml), evaporated to obtain a residue, which was purified by chromatography on silica gel (column: Gemini 150x25 5 μm; mobile phase: water (0.05% ammonium hydroxide, o6./o6.)/ CH ₃ CN: gradient from 42/58 to 12/88, gradient elution time: (min): 10, 100% B retention time (min): 2, flow rate (ml/min): 25).

The required fractions were collected and dried under vacuum to obtain a residue. The residue was lyophilized to give 65 mg (28% yield) of compound 22 as a light yellow solid.

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Example B7.

Receiving connection 24

Intermediate 17 (180 mg, 0.36 mmol) and 3N. NaOH (0.61 ml, 1.82 mmol) in MeOH (10 ml) was stirred at RT. for 1 hour. The mixture was cooled to room temperature, poured into water and extracted with EtOAc. The organic layer was dried over MgSO ₄ , filtered and evaporated to dryness. The residue was lyophilized with 20/80 acetonitrile/water to give 130 mg of compound 24 (79% yield).

Example B8.

Receiving connection 25

At 5°C, 4 N was added dropwise to a solution of intermediate 43 (100 mg, 0.2 mmol) in DCM (10 mL). HCl in dioxane (246 µl, 0.99 mmol) and the mixture was stirred at room temperature. for 15 hours. The reaction mixture was evaporated to dryness. The residue was then taken up with DCM, washed with NaHCO ₃ . The organic layer was dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by chromatography on silica gel (stationary phase: SiOH irregular particles, 15-40 µm, 24 g, mobile phase: NH ₄ OH/DCM/MeOH gradient 0.5/95/5 to 1/90/10) . The purified fractions were collected and evaporated to dryness. The residue was lyophilized with 20/80 acetonitrile/water to give 25 mg of compound 25 (31% yield).

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 25, starting from the corresponding starting materials.

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Connection number Structure Quantity (mg) Exit (%) Compound 37 (from intermediate 44) h ₂ n ⁴ \—o Ϊ ¹ F ₃ C 250 84 Compound 112 (from intermediate 27) Η O G ,_1 /th\ F ₃ CS 35 17

Example B9.

Receiving connection 26

In a sealed tube, 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-th]pyrimidine (0.15 g, 0.594 mmol), 6-(phenylmethoxy)-2-azaspiro[3.3] heptane (0.145 g, 0.713 mmol) and DIPEA (0.205 ml, 1.19 mmol) in isopropanol (2 ml) were heated at 90°C overnight. The solution was cooled to room temperature. and poured into water, then extracted with EtOAc. The organic layer was washed with water, dried over MgSO ₄ , filtered and evaporated to dryness.

The crude product was crystallized from Et ₂ O and dried. The residue was purified by chromatography on silica gel (15-40 µm, 24 g, eluent: heptane/EtOAc: 80/20 to 20/80). The purified fractions were mixed and the solvent was evaporated. The residue was taken up in Et ₂ O, filtered and dried to give 0.111 g of compound 26 (45% yield).

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Example B10.

Receiving connection 28

A mixture of intermediate 10b (200 mg), 1-methyl-1H-pyrazole-4-carboxaldehyde (183 mg; 1.66 mmol) and AcOH (32 μl; 0.555 mmol) in DCE (6 ml) was stirred at 50°C in within 2 hours. The reaction mixture was cooled to room temperature and NaBH(OAc) ₃ (353 mg; 1.665 mmol) was added. The reaction mixture was stirred at room temperature overnight, poured into 10% aqueous K2CO3 and extracted with DCM. The organic layer was decanted, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 24 g; mobile phase: gradient 0% NH4OH, 0% MeOH, 100% DCM to 1% NH4OH, 10% MeOH, 90% DCM). The purified fractions were collected and evaporated to dryness to obtain 165 mg of compound 28 as an oil (73%). Compound 28 was dissolved in ACN and HCl (4N in dioxane) (277 μL; 1.11 mmol) was added. The HCl salt was filtered, and it was found to be too hydroscopic. The residue was then dissolved in DCM/MeOH and the organic layer was washed with 10% aqueous K2CO3, dried over _MgSO4 , filtered and evaporated to dryness. The resulting precipitate was dissolved in ACN and fumaric acid (47 mg; 0.404 mmol; 1 eq.) was added and the solution allowed to stand until crystallized (overnight). The precipitate was filtered, washed with ACN, then EtOAc and dried to give 188 mg of compound 28 as the fumarate salt (1 equivalent based on 1H NMR).

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 28, starting from the corresponding starting materials.

Connection number Structure Quantity (mg) Exit (%) Compound 34 (from intermediate 10b and 1-methyl-1H-pyrazole-3 carboxaldehyde) LL? 147 Compound 44 (from intermediate 10b and isobutyraldehyde) F ₃ C 80

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Example B11.

Receiving connection 29

At 10°C 4 n. HCl in dioxane (0.7 ml; 2.85 mmol) was added to a solution of intermediate 50 (145 mg; 0.28 mmol) in ACN (7 ml). The solution was stirred at room temperature. during the night. The solution was evaporated to dryness. The residue was taken up in ice water, made basic with NH ₄ OH and DCM added. The organic layer was separated, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (stationary phase: SiOH irregular particles, 15-40 μm, 12 g, mobile phase: DCM/MeOH/NH ₄ OH 90/10/10). The purified fractions were collected and evaporated to dryness. The residue was lyophilized with 20/80 acetonitrile/water to give 0.050 g (43% yield) of compound 29.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 29, starting from the corresponding starting materials.

Connection number Structure Amount (mg) Exit (%) Compound 40 (from intermediate 47) n N__. N F ₃ C 2.68HC1 2.1H ₂ O 102 61 Compound 41 (from intermediate 48) h NN Vk NF F ₃ C 72 55

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Example B12.

Receiving connection 36

F ₃ C

In a stream of _N2, 1-isopropyl-1H-pyrazole-4-carbaldehyde (133 mg, 0.68 mmol) and AcOH (51 µl, 0.87 mmol). The mixture was stirred at room temperature. for 2 hours. NaBH(OAc) ₃ (742 mg, 3.5 mmol) was added and the mixture was stirred at room temperature. during the night. The mixture was poured into ice water and separated. The aqueous layer was extracted with DCM. The organic layer was washed with brine, then dried over MgSO ₄ , filtered and evaporated. The residue was purified by chromatography on silica gel (stationary phase: SiOH irregular particle, 15-40 µm, 24 g, mobile phase: DCM/MeOH (+10% NH4OH): gradient 97/3 to 90/10. Purified fractions were collected and evaporated to dryness.The residue was lyophilized with acetonitrile/water: 20/80 to obtain 0.057 g (15%) of compound 36.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 36, starting from the corresponding starting materials.

Connection number Structure Amount (mg) Exit (%) Compound 76 (from intermediate 11 and intermediate 51) LL? 73 33

- 73 045364

Example B13.

Receiving connection 45

To a solution of intermediate 11c (200 mg) in dichloroethane (10 ml) was added 2methylbenzaldehyde (59 mg; 0.494 mmol), NaBH(OAc) ₃ (523 mg; 2.47 mmol) and triethylamine (150 mg; 1.48 mmol). . The mixture was stirred at room temperature overnight and then a saturated aqueous solution of NaHCO ₃ (10 ml) and DCM (10 ml) was added. The mixture was separated and the aqueous layer was extracted with DCM (10 mlx2).

The organic layers were combined, washed with water (10 ml), dried over Na ₂ SO ₄ , evaporated to obtain 300 mg of a yellow oil, which was purified using preparative high performance liquid chromatography (column: Kromasil 150x25 mmx10 μm; conditions:

A: water (0.05% ammonium hydroxide, v/v), B: MeCN, start: A (52%) and B (48%), end: A: (22%) and B (78 %), gradient elution time: (min) 8; flow rate (ml/min) 30.

Fractions containing product were collected and the solvent was evaporated in vacuo. The aqueous layer was lyophilized to dryness to obtain 150 mg (70%) of compound 45 as a white solid.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 45, starting from the corresponding starting materials.

Connection number Structure Quantity (mg) Exit (%) Compound 46 (from intermediate 11c and mtoluylaldehyde) P 5 /X F3C ^Ν salt on 190 Compound 47 (from intermediate 11c and 2methoxy-5methylbenzaldehyde) νΛ- ъ/Л F ₃ C 120

- 74 045364

F 150 F— Compound 48 (from intermediate N__ connections 11c and 2,4- □ difluorobenzaldehyde) F ₃ C / 160 ABOUT Compound 49 (from G intermediate connections 11s and i- O toluylaldehyde) ϊ ^F 3 ^C 155 Connection 50 (of intermediate connections 11c and 2,4- dimethylbenzaldehyde) V //—/Afi F ₃ C SA _K J ^N Salt HC1 166 \ — -* / Compound 54 (from intermediate connections 11c and 2- fluorobenzaldehyde) Ϊ ^F 3 ^CS _CojIbHC1 F A=\ 150 Compound 56 (from G intermediate \ connections 11c and 3- ςς fluorobenzaldehyde) ABOUT /-ΖΎι /\ II I ^N Salt HC1

- 75 045364

Example B14.

Receiving connection 108

A mixture of (S)-5-methyl-5-azaspiro[2.4]heptane-6-carboxylic acid (94 mg; 0.61 mmol), HBTU (231 mg; 0.61 mmol) and DIPEA (0.52 ml; 3 .04 mmol) in DMF (5 ml) was stirred for 1 hour. A solution of intermediate 10b (200 mg) in DMF (5 ml) was then added and the reaction mixture was stirred at room temperature overnight. The reaction mixture was poured into ice water, increased in basicity with 10% aqueous K2CO3 and extracted with EtOAc. The organic layer was washed with H2O, then brine, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 10 g; mobile phase: gradient 3% MeOH, 97% DCM to 10% MeOH, 90% DCM). Fractions containing product were collected and evaporated to dryness to obtain 144 mg of crude fraction 1. A second purification was performed (SiOH irregular, 40 g; mobile phase: 0.5% NH4OH, 95% DCM, 5% MeOH). Fractions containing product were collected and evaporated to dryness to obtain 43 mg of crude fraction 2.

Fraction 2 was again purified by silica gel chromatography (SiOH irregular particle, 10 g; mobile phase: gradient 3% MeOH, 97% DCM to 10% MeOH, 90% DCM). Fractions containing the product were collected and evaporated to dryness. The resulting precipitate was taken up with diisopropyl ether. The solid was filtered and dried to obtain 17 mg of compound 108.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 108, starting from the corresponding starting materials.

Connection number Structure Quantities about (mg) Exit (%) Compound 74 (from intermediate 32 and Nisopropylethylenediamine) ^NH HN y=° AA— ⁴ Ν—. ъ F ₃ CS 10 7 Compound 75 (from intermediate 32 and 2 aminoethanol) OH HN Y=o (A ъ αΛ f ₃ c 22 13

- 76 045364

Example B18

Receiving connection 57

LiAlH ₄ (66 mg, 1.73 mmol) was added to intermediate 54 (450 mg, 0.693 mmol) in THF (12 ml). The reaction mixture was stirred at room temperature. for 1.5 hours. The reaction mixture was quenched with saturated aqueous NH ₄ Cl, extracted with DCM and concentrated to give a white solid. This solid was purified using preparative high performance liquid chromatography (column: Xtimate C18 150x25 mmx10 µm, conditions: water (0.05% ammonium hydroxide, o6./o6.)/ACN: gradient 52/48 to 42/58). 0.1 ml of 1 N was added to the aqueous layer. HCl. The solution was lyophilized to obtain 30 mg of compound 57 as a yellow solid (HCl salt).

Example B19.

Receiving connection 58

To a solution of intermediate 8 (200 mg, 0.5 mmol) in THF (10 mL) was added isobutyraldehyde (70 μL, 0.77 mmol) and TEA (0.37 mL, 2.63 mmol). The mixture was stirred at room temperature. for 3 hours. NaBH(OAc) ₃ (317 mg, 1.5 mmol) was added and the solution was stirred at room temperature. during the night. The solution was poured into cooled water and its basicity was increased with powdered K2CO3. The product was extracted with EtOAc. The organic layer was dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (stationary phase: unmodified irregular silica, 40 g, mobile phase: NH ₄ OH/DCM/MeOH: 0.2/98/2). The residue was dissolved in 5 ml of ACN, 2 equiv was added dropwise at 10°C. 4 n. HCl in dioxane (117 µl; 0.47 mmol).

Et ₂ O was added and after 30 minutes the solution was evaporated to dryness, Et ₂ O was added and the precipitate was filtered and dried to obtain 38 mg of compound 58 (HCl salt).

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 58, starting from the corresponding starting materials.

Connection number Structure Amount (mg) Exit (%) Compound 59 (from intermediate 8 and 1methyl-1H-pyrazole-4carbaldehyde) § xL ^F ' ^CSN Salt HCl 73

- 77 045364

Example B21.

Receiving connection 62

M TBAF in THF (0.815 ml, 0.815 mmol) was added dropwise to a solution of intermediate 55 (0.248 g, 0.407 mmol) in Me-THF (8 ml) and the reaction mixture was stirred at RT. during the night. The reaction mixture was poured into 10% aqueous K ₂ CO ₃ solution and extracted with EtOAc. The organic layer was washed with 10% aqueous K ₂ CO ₃ (30 ml), water (30 ml) and brine (30 ml), dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by chromatography on silica gel (80 g, 15-40 µm, eluent: DCM/MeOH: 97/3 to 88/12). The purified fractions were mixed and the solvent was evaporated to obtain 0.043 g of compound 62 (21% yield).

Example B22.

Receiving connection 63

2,2-Dimethyltetrahydropyran-4-carbaldehyde (87 mg; 0.609 mmol) and NaBH(OAc) ₃ (645 mg; 3.045 mmol) were added at RT. to a solution of intermediate 11 (200 mg; 0.609 mmol) in DCE (4 ml) and the reaction mixture was stirred overnight. The reaction mixture was diluted with DCM and poured into a 10% aqueous K ₂ CO ₃ solution. The organic layer was decanted, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 24 g; mobile phase: NH ₄ OH/MeOH/DCM: gradient 0/0/100 to 0.7/7/93). The purified fractions were collected and evaporated to dryness. The residue was lyophilized from water/ACN (80/20; 10 ml) to give 155 mg of compound 63 (56% yield).

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 63, starting from the corresponding starting materials.

Connection number Structure Amount (mg) Exit (%) Compound 65 (from intermediate I and 3methyloxetane-3 carbaldehyde) O zk Г—О ^ω у / \ / /—' ^ζ 1— ζ λ—ζ 1 \— _ζ Ν— 71 60

- 78 045364

Example B23.

Receiving connection 64

Intermediate mixture

(150 mg, 0.457 mmol), 2,2,2-trifluoroethyl trifluoromethanesulfonate (69 µl, 0.502 mmol) and DBU (CAS[6674-22-2]) (136 µl, 0.914 mmol) in DMSO (3 ml) were stirred at k.t. for 18 hours. The reaction mixture was poured into water and extracted with EtOAc. The organic layer was washed several times with water, then with brine, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (SiOH irregular particle, 24 g; mobile phase: NH4OH/MeOH/DCM gradient 0/0/100 to 0.7/7/93). The purified fractions were collected and evaporated to dryness. The residue was purified a second time by chromatography on silica gel (SiOH irregular particle, 10 g; mobile phase: EtOAc/heptane: gradient 60/40 to 80/20). The purified fractions were collected and evaporated to dryness. The residue was crystallized from DIPE to obtain, after drying under vacuum at 50°C, 100 mg of compound 64 (53% yield).

Example B24.

Receiving connection 67

Under _N2 atmosphere, a solution of intermediate 11 (100 mg; 0.31 mmol), 2-(tetrahydro-2Hpyran-4-yl))acetaldehyde (48 μl; 0.37 mmol) in THF (3 ml) was stirred at .T. for 3 hours. NaBH(OAc) ₃ (129 mg, 0.61 mmol) was added and the mixture was stirred at room temperature. during the night. A 10% aqueous solution of K ₂ CO ₃ and EtOAc was added. The mixture was extracted with EtOAc (x3). The organic layers were combined, washed with brine, then dried over MgSO ₄ , filtered and the solvent was evaporated. The residue (136 mg) was purified by silica gel chromatography (SiO ₂ , 4 g; gradient: 95% DCM, 5% MeOH, 0.5% NH4OH to 90% DCM, 10% MeOH, 1% NH4OH). Fractions containing the product were collected and the solvent was evaporated to give 90 mg of a colorless oil, which was recrystallized with diisopropyl ether. The precipitate was filtered and dried to obtain 45 mg (34%) of compound 67 as a white solid.

- 79 045364

Example B25.

Receiving connection 71

In a sealed tube under N2 atmosphere, intermediate 35 (211 mg; 1.37 mmol) and Ti(OiPr) ₄ (436 μl; 1.83 mmol) were added to a solution of intermediate 11 (300 mg; 0.914 mmol) in THF ( 6 ml). The solution was stirred at 50°C for 5 hours, then at room temperature. during the night. The reaction mixture was cooled to 5°C and 2 M isopropylmagnesium chloride in THF (2.28 ml; 4.57 mmol) was added dropwise. The reaction mixture was slowly heated to room temperature. and stirred overnight. The reaction mixture was diluted with EtOAc and poured into 10% aqueous K2CO3 solution. The precipitate was removed by filtration through Celite®. The organic layer was decanted, washed with brine, dried over MgSO4, filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 40 g; mobile phase: gradient 0% MeOH, 100% DCM to 10% MeOH, 90% DCM). Fractions containing the product were collected and evaporated to dryness to obtain 0.337 g of a residue representing the intermediate, which was again purified by reverse phase chromatography (stationary phase: YMC-actus Triart C18 10 µm, 30x150 mm, mobile phase: gradient from 55% NH4HCO3 0.2%, 45% ACN to 0% NH4HCO3 0.2%, 100% ACN). The purified fractions were collected and evaporated to dryness. The residue was lyophilized with 20/80 acetonitrile/water to give 120 mg (26%) of compound 71.

Example B26.

Receiving connection 77

Hydrazine (36 μL, 0.92 mmol) was added to a solution of intermediate 41 (110 mg, 0.18 mmol) in ethanol (5 mL). The solution was heated at 70°C for 1 hour 30 minutes. The reaction mixture was cooled to RT, then poured into water and extracted with DCM. The organic layer was washed with brine, then dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (stationary phase: unmodified irregular silica, 40 g, mobile phase: NH ₄ OH/DCM/MeOH: 1/85/15).

The purified fractions were collected and evaporated to dryness. The residue was dissolved in 2 ml of ACN, 3 equiv was added dropwise at 10°C. 6 n. HCl in iPrOH. Et ₂ O was added and after 30 minutes the precipitate was filtered and dried to obtain 83 mg of compound 77 (yield 37%).

- 80 045364

Example B28.

Receiving connection 82

TFA (1.5 ml) was added to a solution of intermediate 25 (300 mg, 0.571 mmol) in DCM (15 ml) and the reaction mixture was stirred for 18 hours. The reaction mixture was poured into 10% aqueous K ₂ CO ₃ solution and extracted with using DCM. The organic layer was decanted, filtered through Chromabond® and evaporated to dryness. The residue was purified by silica gel chromatography (SiOH irregular particle, 10 g; mobile phase: NH4OH/MeOH/DCM: gradient 0.3/3/97 to 1.5/15/85). The purified fractions were collected and evaporated to dryness. The residue was crystallized from DIPE and dried to obtain 118 mg of compound 82 (48% yield).

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 82, starting from the corresponding starting materials.

Connection 315 (from intermediate 109) \ /--\ ^NH \—/rs \'' N— Λ 140 52 Connection 318 (from intermediate 112 and at T = 0°C) n /---\ ^RS g-----( N-___ / 'GT 1 salt HC1 F ₃ C 200

- 81 045364

Example B29.

Receiving connection 84

Intermediate 11b (198 mg) and 4-fluorophenylacetone (68 µl, 0.51 mmol) in THF (5 ml) were stirred at RT. during the night. NaBH(OAc) ₃ (161 mg, 0.76 mmol) was then added in portions. The mixture was stirred at room temperature. for 24 hours. The solution was poured into cooled water and its basicity was increased with a 3 N solution. NaOH, EtOAc was added. The organic layer was separated, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (stationary phase: unmodified irregular silica, 40 g, mobile phase: NH ₄ OH/DCM/MeOH: 0.1/97/3). The purified fractions were collected and the solvent was evaporated under vacuum. The residue was lyophilized with 20/80 acetonitrile/water to give 30 mg of compound 84.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 84, starting from the appropriate starting materials.

Connection number Structure Amount (mg) Exit (%) Connection 307 (from intermediate 11b and 1-(4fluorophenyl)-3methylbutan-2-one) F Ul * '---(RS ъ F ₃ C 46

Example B32.

Receiving connection 133

- 82 045364 connection 137 and connection 138

Ti(OEt) ₄ (251 μl; 1.2 mmol) was added at room temperature to a solution of intermediate 11 (200 mg; 0.6 mmol) and 2,5,6,7-tetrahydro-2-methyl-4H-indazole -4-one (120 mg; 0.8 mmol) in dichloroethane (5 ml) and MeOH (1.5 ml). The mixture was stirred at room temperature. for 2 hours, then NaBH ₃ CN (127 mg; 2 mmol) was added in portions. The mixture was stirred at room temperature for 2 days. The solution was poured into cooled water and DCM was added. The mixture was made more basic with powdered K ₂ CO ₃ and filtered through a Celite® pad. The product was extracted with DCM. The organic layer was dried over MgSO ₄ , filtered and evaporated to dryness.

The residue (361 mg) was purified by silica gel chromatography (stationary phase: unmodified irregular silica, 40 g, mobile phase: 0.5% NH4OH, 95% DCM, 5% MeOH). The product containing fraction was mixed and concentrated to give 120 mg (43%) of compound 133.

Chiral resolution of compound 133 was performed by chiral SFC (stationary phase: Chiralpak AD-H, 5 µm, 250x30 mm, mobile phase: 70% CO2, 30% MeOH (0.3% iPrNH ₂ )). Fractions containing the products were mixed and concentrated to yield mg of fraction 1, which was lyophilized with 20/80 acetonitrile/water to yield 40 mg (43%) of compound 137 as a white powder.

mg of fraction 2, which was lyophilized with a mixture of 20/80 acetonitrile/water to obtain 42 mg (46%) of compound 138 as a white powder.

Example B33.

Receiving connection 145

- 83 045364 connection 154 and connection 155

A solution of 1-tetrahydro-2H-pyran-4-ylethanone (351 mg; 2.74 mmol), intermediate 11 (600 mg; 1.83 mmol), Ti(OiPr) ₄ (870 μl; 2.92 mmol) in EtOH (3 ml) was stirred for 2 hours at 45°C. Additional EtOH (18 ml) and NaBH ₄ (138 mg; 3.65 mmol) were added. The reaction mixture was stirred at room temperature for 5 hours. The reaction mixture was diluted with DCM and poured into a 10% aqueous K ₂ CO ₃ solution. Insoluble matter was removed by filtration through Celite®. The organic layer was separated, washed with water, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 24 g; mobile phase: gradient 0% NH4OH, 0% MeOH, 100% DCM to 1% _NH4Oh , 10% MeOH, 90% DCM). Fractions containing product were collected and evaporated to dryness to obtain 487 mg (60%) of compound 145. Enantiomers of compound 145 were separated using chiral SFC (CHIRALPAK AD-H 5 µm, 250x30 mm; mobile phase: 70% CO2, 30% mixtures 50/50 EtOH/iPrOH, v/v). Fractions containing products were collected and evaporated to dryness. The residues were lyophilized from water/ACN (80/20; 10 ml) to obtain 171 mg (21%) of compound 154 and 178 mg (22%) of compound 155.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 145, starting from the corresponding starting materials. The most significant small deviations are indicated in the Connection Number column.

- 84 045364

Connection number Structure Quantity Exit D Compound 279 (from intermediate 11 and 1 -(3-methyloxetan-3 yl)ethenone [136338104-7]). With 6 eq. Ti(iPrO)4 and 2 eq. NaBH4 ^rs G σ F ₃ C 243 mg 37% Compound 158 and Compound 159. From SFC purification of intermediate 158 (CHIRACEL OJ-H 5 µm, 250*30 mm, mobile phase: 80% CO ₂ , 20% EtOH (0.3% iPrNH ₂ )) Λϋ __Ν 1—о rYTj F ₃ C Compound 158 77 mg 12%

- 85 045364

YA F ₃ C Connection 159 80 mg 12% Compound 192 (from intermediate 11 and 1(tetrahydro-2,6dimethyl-2[/-pyran-4yl)-1-propanone) C 1.6 eq. Ti(iPrO)4 and 2 eq. NaBH ₄ \___ ιΥγ-A3 co L« 76 mg 14% Compound 280 (from intermediate 11b and 3methyl-1-(tetrahydro2//-pyran-4-yl)-1 butanone). With 6 eq. Ti(iPrO)4 and 2 eq. NaBH ₄ /—\ ° J RS LJ A F ₃ C 195 mg Compound 179 and Compound 180. From SFC purification of Compound 280 (CHIRALPAK AD-H 5 µm, 250*30 mm; mobile phase: 75% CO ₂ , 25% MeOH (0.3% iPrNH ₂ )) Yo __N ^R '---/ $ Yj F ₃ C S-^ _N ^ Compound 179 70 mg 10%

- 86 045364

/—L ° NrL” F ₃ C S-^ _N ^ Compound 180 69 mg 10% Compound 250 C 6 eq. Ti(iPrO)4 and 2 eq. NaBH ₄ Compound 163 and o .— ---b RS N— Yo F ₃ C Compound 250 From intermediate compound I and 1-(1,1dioxotetrahydro-2Nthiopyran-4-yl)ethenone o ......р ъ ΗΊΟ F ₃ C ^S ^ N^ Compound 163 369 mg 123 mg 62% 21%

- 87 045364

connection 164 o V° -/*s А F ₃ C Compound 164 From chiral SFC compound 250: CHIRACEL OJ-H 5 µm, 250*30 mm; mobile phase: 70% CO ₂ , 30% MeOH (0.3% iPrNH ₂ ) 123 mg 21% Connection 314 (from intermediate 11 and 4cyanophenyl acetone) X A F ₃ C 105 mg 15%

Preparation of compound 312 (diastereoisomer A (mixture of 2 compounds (RR and SS) or (RS and SR)) and compound 313 (diastereoisomer B (mixture of 2 compounds (RS and SR) or (RR and SS)).

Compound 312. Diastereomer A (RR and SS) or (RS and SR).

Reaction mixture 1. Solution of 3-methyl-1-(6-oxaspiro[4.5]dec-9-yl)-1-butanone (1.5 eq), intermediate 11 (100 mg; 0.285 mmol), Ti(OiPr ) ₄ (1.6 eq.) in ethanol (0.25 ml) was stirred for 2 hours at 45°C. Ethanol (3 ml) was added and NaBH ₄ (2 eq.) was added. The reaction mixture was stirred at room temperature for 18 hours.

Reaction mixture 2. Solution of 3-methyl-1-(6-oxaspiro[4.5]dec-9-yl)-1-butanone (546 mg; 2.436 mmol; 2 eq.), intermediate 11 (400 mg; 1.22 mmol), Ti(OiPr)4 (580 µl; 1.95 mmol) in ethanol (1 ml) was stirred for 2 hours at 45°C. Ethanol (12 ml) was added and NaBH ₄ (92 mg; 2.436 mmol) was added. The reaction mixture was stirred at room temperature for 18 hours.

The two reaction mixtures were combined and diluted with EtOAc, poured into 10% aqueous K ₂ CO ₃ solution and filtered through a Celite® pad. The organic layer was decanted, washed with brine, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 24 g; mobile phase: gradient 0% NH4OH, 0% MeOH, 100% DCM to 0.9% NH4OH, 9% MeOH, 91% DCM). The required fractions were collected and evaporated to dryness.

- 88 045364

The residue (280 mg) was further separated by silica gel chromatography (irregular SiOH, 24 g; mobile phase: 67% heptane, 33% EtOAc (+ 5% MeOH containing 10% NH ₄ OH)). The required fractions were collected and evaporated to dryness to obtain:

100 mg (12%) of compound 312 (eluted first; not sufficiently pure), which was further purified by silica gel chromatography (irregular SiOH, 24 g; mobile phase: gradient from 0% NH ₄ Oh, 0% MeOH, 100 % DCM to 0.5% NH4OH, 5% MeOH, 95% DCM). The purified fractions were collected and evaporated to dryness. Purified fractions were lyophilized from water/ACN (80/20; 10 ml) to obtain 80 mg (10%) of compound 312.

110 mg (13%) of compound 313 (eluted second; not sufficiently pure), which was further purified by silica gel chromatography (SiOH irregular, 24 g; mobile phase: gradient of 0% NH4OH, 0% MeOH, 100% DCM up to 0.5% NH4OH, 5% MeOH, 95% DCM). The purified fractions were collected and evaporated to dryness. Purified fractions were lyophilized from water/ACN (80/20; 10 ml) to obtain 70 mg (9%) of compound 313.

Example B34.

Receiving connection 147

A mixture of intermediate 60a (216 mg; 0.4 mmol) and TFA (1 mL; 13.067 mmol) in DCM (10 mL) was stirred at RT. for 4 hours. The reaction mixture was diluted with DCM and increased in basicity with 10% aqueous K ₂ CO ₃ solution. The organic layer was decanted, washed with water, filtered through Chromabond® and evaporated to dryness. The residue (200 mg) was purified by silica gel chromatography (irregular SiOH, 10 g; mobile phase: 85% DCM, 14% MeOH, 1% NH4OH). The purified fractions were collected and evaporated to dryness. The residue was lyophilized from water/ACN (80/20; 10 ml) to give 136 mg of compound 147 (77%).

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 147, starting from the appropriate starting materials.

- 89 045364

Connection number Structure Quantity B.O. Exit D Compound 146 (from intermediate 60b) /—\ ' ^NH % F ₃ C 130 mg 71% Connection 150 (from intermediate connection 63 a) )--\ ^NH if F ₃ C 29 mg 45% Compound 148 (from intermediate 68) /—\ ^NH V RS \--/ I salt TFA HTj F ₃ C 50 mg Compound 168 (from intermediate 68a) _7 }---( NH 7 >20 45 mg 50% Compound 169 (from intermediate 68b) /---\ ' ^NH o+o F ₃ C 50 mg 58%

Example B35.

Receiving connection 156

- 90 045364 connections 193 connections 162 and connections 157

A solution of 2-methyl-1-(tetrahydro-2H-pyran-4-yl)-1-propanone (1.66 g, 10.63 mmol) in THF (30 ml) was added to a solution of intermediate 11 (2.33 g ; 7.08 mmol) and TFA (3.3 ml; 42.5 mmol) in THF (45 ml). The reaction mixture was stirred at room temperature. during the night. NaBH(OAc) ₃ (4.5 g, 21.25 mmol) was then added in portions. The reaction mixture was stirred at room temperature. within 7 days.

The reaction mixture was stirred at room temperature. within 3 days. The solution was poured into a 10% aqueous solution of K ₂ CO ₃ and EtOAc was added. The mixture was extracted with EtOAc (3x). The organic layers were combined, washed with brine, dried over MgSO ₄ , filtered and the solvent was evaporated. The residue (3.9 g; yellow oil) was purified by chromatography on silica gel (SiO ₂ ; 40 g; eluent: 97% DCM, 3% MeOH, 0.3% NH4OH to 90% DCM, 10% MeOH, 1% NH4OH ). The purified fractions were collected and the solvent was evaporated to give 666 mg of a pale brown solid as compound 156.

Compound 156 was purified using reverse phase (YMC-actus Triart C18 10 µm, 30x150 mm, mobile phase: gradient from 50% NH4HCO3 0.2%, 50% ACN to 0% NH4HCO3 0.2%, 100% ACN). The purified fractions were collected and the solvent was evaporated to obtain 66 mg of compound 157 (colorless oil) and 264 mg of compound 156 (8%; colorless oil).

mg of compound 156 was lyophilized with water-ACN to obtain 47 mg of compound 156 (white solid).

Compound 157 was lyophilized with water-ACN to obtain 53 mg of compound 157 (2%, white solid).

Compound 156 (214 mg) was purified using chiral SFC (CHIRALPAK AD-H 5 µm, 250x30 mm, mobile phase: 75% CO2, 25% EtOH (0.3% iPrNH ₂ )). The purified fractions were collected and the solvent was evaporated to obtain 82 mg of compound 193 (colorless oil) and 82 mg of compound 162 (colorless oil).

Compound 193 was lyophilized with water-ACN to obtain 72 mg of compound 193 (2%, white solid).

- 91 045364

Compound 162 was lyophilized with water-ACN to obtain 77 mg of compound 162 (2%, white solid).

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 193, starting from the corresponding starting materials.

Connection number Structure Quantity Exit d Compound 309 as mixture 4 diastereoisomers (ratio: 65/35) (from intermediate 11 and 105) \ RS/ -о rs)—< 1 Λ F ₃ C 10 mg 7%

Example B36.

Receiving connection 161

connections 166 and connections 167

In a sealed tube under _N2 , 1,3-dimethyl-1H-pyrazole-4-carbaldehyde (284 mg; 2.28 mmol) and Ti(iPrO) ₄ (727 μL; 3.05 mmol) were added to the solution of the intermediate 11 (500 mg; 1.52 mmol) in THF (10 ml). The solution was stirred at 50°C for 2 hours. The reaction mixture was cooled to 5°C and iPrMgCl (3.8 ml; 7.61 mmol) was added dropwise. The reaction mixture was slowly heated to room temperature. and stirred overnight. The reaction mixture was poured into 10%

- 92 045364 aqueous solution of K ₂ CO ₃ and EtOAc. The insoluble material was filtered through a Celite® pad, then the organic layer was decanted, dried over MgSO ₄ , filtered and the solvent was evaporated. The residue (866 mg, brown oil) was purified by silica gel chromatography (SiO2; 40 g; eluent: 96% DCM, 4% MeOH, 0.4% NH4OH to 93% DCM, 7% MeOH, 0.7% NH4OH) . The purified fractions were collected and the solvent was evaporated to dryness. The residue (496 mg, yellow oil) was recrystallized with diethyl ether. The precipitate was filtered and dried to obtain 324 mg of compound 161 (45%, white solid).

270 mg of compound 161 (purified using chiral SFC (CHIRALPAK AD-H 5 µm, 250x30 mm, mobile phase: 70% CO2, 30% iPOH (0.3% iPrNH2)). Purified fractions were collected and solvent evaporated to give 128 mg compound 166 (18%, colorless oil) and 131 mg of compound 167 (18%, colorless oil).

Compound 166 was lyophilized with water-ACN to obtain 110 mg of compound 166 (15%, white solid).

Compound 167 was lyophilized with water-ACN to obtain 115 mg of compound 167 (16%, white solid).

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 161, starting from the corresponding starting materials. The most significant small deviations are indicated in the Output column.

Connection number Structure Quantity B.O. Exit Compound 172 (from intermediate 11 and 1,5dimethyl-1H-pyrazole4-carbaldehyde) \ )-WITH . / RS \^N σ' xrS F ₃ C 26 mg 4% Compound 173 and Compound 174. From SFC purification of Compound 172 (CHIRALCEL OD-H 5 µm, 250*30 mm, mobile phase: 80% CO ₂ , 20% EtOH (0.3% iPrNH ₂ )) /.......h Vn XiS F ₃ C Connection 173 )-¾ Г AIj F ₃ C SY _n A Connection 174 109 mg 105 mg 15% 14% Compound 186 (from intermediate 70b and 1methyl-1//-pyrazole-4carboxaldehyde) /%—\ \ / RS \=\ !—/ N /V ^s % ^z Xi F ₃ C SA _n A 31 mg 9%

- 93 045364

\/* ^R 53 mg 15% Compound 188 and Compound 189. From SFC purification of Compound 186 (CHIRALPAK AD-H 5 µm, 250*30 mm, mobile phase: 85% CO ₂ , 15% EtOH (0.3% iPrNH ₂ )) /—V ^- ' LN / V ^s % ^z F ₃ C Connection 188 /—LN / \ * ^s % ^z F ₃ C Connection 189 74 mg 21% Connection 281 From intermediate 70a and 1methyl-17/-pyrazole-4carboxaldehyde KG F ₃ C 250 71% Ti(OEt) ₄ was used in the synthesis. Connection 190 and connection 191. N 77 mg 22% From purification with SFC compound 281 (CHIRALPAK AD-H 5 µm, 250*30 mm, mobile phase: 85% CO ₂ , 15% EtOH (0.3% iPrNH ₂ )) HTj F ₃ C Connection 190 ΙΨ F ₃ C 61 mg 17% Connection 191

Example B38.

Receiving connection 170

F

Intermediate 11b (198 mg) and 1-(3,5-difluorophenyl)propan-2-one) (86 mg; 0.51 mmol) in THF (5 ml). The mixture was stirred at room temperature. during the night. NaBH(OAc) ₃ (161 mg, 0.76 mmol) was then added in portions. The mixture was stirred at room temperature. for 24 hours. The solution was poured into chilled water,

- 94 045364 increased its basicity using a 3 N solution. NaOH, EtOAc was added. The organic layer was separated, dried over MgSO ₄ , filtered and evaporated to dryness. The residue (141 mg) was purified by silica gel chromatography (unmodified irregular silica, 40 g, mobile phase: 0.1% NH4OH, 97% DCM, 3% MeOH). The purified fractions were collected and the solvent was evaporated. The residue (50 mg) was lyophilized with 20/80 ACN/water to give 16 mg of compound 170.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 170, starting from the corresponding starting materials.

Connection number Structure Quantity B.O. Exit D Compound 171 (from intermediate 11b and 2methoxyphenylacetone) 1 (t 1 « 1 ^α o ^w 35 mg Compound 183 (from intermediate 11b and 3fluorophenylacetone) Compound 182 (from intermediate 11b and 3methyl-1-phenyl-2butanone) Compound 185 (from intermediate 70a and tetrahydropyran-4carbaldehyde [5067518-8]) F P 1 salt HC1 hTj ⁿ F ₃ C and F ₃ C O O' ^R F ₃ C 38 mg 55 mg 143 mg 56%

- 95 045364

Example B39.

Receiving connection 184

A mixture of intermediate 70a (250 mg; 0.73 mmol), isobutyraldehyde (200 μL; 2.19 mmol) and AcOH (42 μL; 0.73 mmol) in DCE (8 ml) was stirred at 50 °C for 3 h The reaction mixture was cooled to room temperature. and NaBH(OAc) ₃ (464 mg; 2.19 mmol) was added. The reaction mixture was stirred at room temperature. overnight, poured into 10% aqueous K2CO3 solution and extracted with DCM. The organic layer was decanted, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 24 g; mobile phase: gradient 100% DCM to 10% MeOH (+10% NH ₄ OH), 90% DCM). The purified fractions were collected and evaporated to dryness. A hydrochloride salt was obtained. The residue (110 mg, 38%) was dissolved in ACN and 4N was added. HCl in 1,4-dioxane (2 eq.). The solution was evaporated to dryness and absorbed several times with ACN. The residue was crystallized from Et ₂ O to obtain 120 mg of compound 184 (HCl salt).

The compound shown in the table below was prepared by using a method similar to that described for the preparation of compound 184, starting from the appropriate starting materials.

- 96 045364

Example B40.

Receiving connection 289

A solution of intermediate 70a (100 mg; 0.29 mmol), p-toluylaldehyde (50 μL; 0.35 mmol) in dichloroethane (3 mL) was stirred at RT. for 3 hours. NaBH(OAc) ₃ (124 mg, 0.58 mmol) was added and the mixture was stirred at room temperature. during the night. A 10% aqueous solution of K ₂ CO ₃ and DCM was added. The organic layer was decanted, filtered through Chromabond® and evaporated to dryness. The residue was purified by chromatography on silica gel (SiOH irregular particle, 10 g; mobile phase: gradient from 0% NH ₄ OH, 0% MeOH, 100% DCM to 0.5% NH ₄ OH, 5% MeOH, 95% DCM ). The purified fractions were collected and evaporated to dryness. The residue was lyophilized from water/ACN (80/20; 10 ml) to give 73 mg (56%) of compound 289.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 289, starting from the appropriate starting materials. The most significant small deviations are indicated in the Output column.

- 97 045364

Connection number Structure Quantity Exit Compound 290 (from intermediate 70b and ptoluylaldehyde) ζ^ ^z —λ ^ζ —\ ' ^ζ ω 85 mg 65 Compound 291 (from intermediate 70b and 4fluorobenzaldehyde) F Ο A 89 mg 68% THF as solvent Compound 292 (from intermediate 70a and 4fluorobenzaldehyde) hr 86 mg 65% THF as solvent

- 98 045364

Compound 293 (from intermediate 70a and 3methylbutyraldehyde) Λ Ν Λ 68 mg 56% THF in quality e solvent Compound 294 (from intermediate 70b and 3methylbutyraldehyde) F f' Z^N— \s* / 'f 72 ΜΓ 60% THF as solvent Compound 295 (from intermediate 70a and 1methyl-1H-pyrazole-4carbaldehyde) F F Z^\ I Λ \__,z---< & 50 ΜΓ 39% THF as solvent Compound 296 (from intermediate 70a and phenylacetaldehyde) F/ F z.___ XS ^w 50 ΜΓ 38% THF as solvent

- 99 045364

Compound 297 (from intermediate 70b and phenylacetaldehyde) ω xy ^z CV\ iq 'N / 1.8НСО,Н 18 mg eleven% THF as solvent Connection 298 (from VV ^F 20 mg 14% intermediate Du connections 70a and 2-(4- ABOUT fluorophenyl)acetaldehyde Ida) r /—\JL J N ABOUT 74 mg 58% Connection 299 (from at intermediate /S/ THF in connections 70b and 1- WHAT qualities methyl-1H-pyrazole-4- e carbaldehyde) _____// || >N solution x\D Λ owner

Example B41.

Receiving connection 273 connections 200

- 100 045364 and connections 222

A mixture of intermediate 71 (546 mg; 1.01 mmol) and TFA (1.5 ml; 19.6 mmol) in DCM (15 ml) was stirred at RT. within 4 hours. The mixture was evaporated to dryness.

The residue was taken up with DCM and H ₂ O, then increased in basicity with aqueous 3N. NaOH. The organic layer was decanted, dried over MgSO ₄ , filtered and evaporated to dryness to obtain 400 mg (90%) of compound 222.

Enantiomers were separated using chiral SFC (Chiralpak AD-H 5 µm, 250x30 mm; mobile phase: 50% CO2, 50% EtOH (0.3% iPrNH ₂ )). The purified fractions were collected and evaporated to dryness to obtain 106 mg (24%) of compound 273 and 130 mg (29%) of compound 200.

Compound 273 can also be prepared from intermediate 71a using the same procedure.

Compound 200 can also be prepared from intermediate 71b using the same procedure.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 273, starting from the appropriate starting materials.

- 101 045364

Connection number Structure Quantity Exit Connection 211 n ъ A From intermediate 77a 125 mg 56% Connection 213 n '---CS ъ F ₃ C From intermediate 77b 123 mg 55% Connection 259 n /—\ ^RS ъ F ₃ C From intermediate connection 80 96 mg 49%

- 102 045364

Example B42.

Receiving connection 274

- 103 045364 and connections 203

In an N2 atmosphere at room temperature 1-Methyl-1H-pyrazole-4-carbaldehyde (100 mg; 0.913 mmol) and Ti(OEt) ₄ (0.25 ml; 1.233 mmol) were added to a solution of intermediate 11 (200 mg; 0.609 mmol) in THF (3 ml). The solution was stirred at room temperature for 20 hours. The reaction mixture was cooled to 5°C and 2 M iPrMgCl in THF (1.5 ml; 3.045 mmol) was added dropwise. The reaction mixture was stirred for 30 min at 5°C, and slowly warmed to room temperature. for 6 hours and poured into a cold aqueous K2CO3 solution. DCM was added and the reaction mixture was filtered through a Celite® pad. The insoluble material was washed several times with DCM. The organic layer was decanted, filtered through Chromabond® and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 24 g; mobile phase: gradient 0% NH4OH, 0% MeOH, 100% DCM to 0.8% NH4OH, 8% MeOH, 92% DCM). The purified fractions were collected and evaporated to dryness to obtain 195 mg (69%) of the racemic compound. Enantiomers were separated using chiral SFC (Lux Cellulose-25 µm, 250x30 mm; mobile phase: 55% CO2, 45% MeOH (0.3% iPrNH ₂ )). Two fractions were lyophilized from water/ACN (80/20; 12 ml) to obtain 80 mg (28%) of compound 274 and 82 mg (29%) of compound 203.

Example B43.

Receiving connection 197

connections 198 and connections 199

In an N2 atmosphere at room temperature 1-Methyl-1H-pyrazole-4-carbaldehyde (99 mg; 0.9 mmol) and Ti(OEt) ₄ (0.25 ml; 1.2 mmol) were added to a solution of intermediate 11 (197 mg; 0.6 mmol) in THF (3 ml), dis

- 104 045364 the solution was stirred at room temperature. within 20 hours. The reaction mixture was cooled to 0°C and CH ₃ MgBr (3 M in EtOAc; 1 ml; 3 mmol) was added dropwise. The solution was stirred for 30 min at 0°C and its temperature was allowed to slowly rise to room temperature. within 6 hours. The solution was poured into a mixture of cold water and an aqueous saturated solution of NH ₄ Cl, then EtOAc was added. The mixture was filtered through a Celite® pad and extracted with EtOAc. The organic layer was dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 24 g; mobile phase: gradient 0% NH4OH, 0% MeOH, 100% DCM to 0.5% NH4OH, 5% MeOH, 95% DCM). Purified fractions were collected and evaporated to dryness to yield 130 mg (50%) of compound 197. Enantiomers were separated using chiral SFC (Lux Cellulose-4 5 µm 250x21.2 mm; mobile phase: 60% CO2, 40% MeOH (+0 .3% iPrNH2)). Purified fractions were collected, evaporated and lyophilized from water/ACN (80/20) to obtain 46 mg (17%) of compound 198 and 45 mg (17%) of compound 199.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 274, starting from the appropriate starting materials.

Connection number Structure Quantities 0 Exit d Compound 214 7% Λ/Ο F ₃ CS From intermediate 11, 1isopropyl-1H-pyrazole-4-carbaldehyde and 2 M iPrMgCl in THF 19 mg 13% Connection 283, ъ F ₃ C Compound 283 From the intermediate compound methyl 1 H-pyrazole-4-carbaldehyde EtMgBr in THF 11, 1 and 1 M 143 mg 52% connection 220 / I ?—< _n rCu F ₃ C 58 mg 21%

- 105 045364

And Connection 220 61 mg 22% connection 221 ъ F ₃ C Compound 221 From purification of compound 283 using chiral SFC: CHIRALPAK IC 5 µm, 250*21.2 mm; mobile phase: 60% CO2, 40% EtOH (0.3% iPrNH ₂ ) Compound 223 Η Ν O - F ₃ C From intermediate 11, tert-butyl-4-formyl-1H-pyrazole-1 carboxylate and 3 M MeMgBr in Et ₂ O 28 mg 20% Compound 228 S 0 —<RS ъ F ₃ C From intermediate 11, isothiazole-4-carbaldehyde and 3 M 77 mg 57%

- 106 045364

MeMgBr in Et ₂ O Compound 230, MiA. / ъ F ₃ CS Compound 230 From intermediate I, 1methyl-1H-pyrazole-4-carbaldehyde and 2 M isobutylmagnesium bromide in Et ₂ O / ^{1 R} 343 mg 78% connection 247 ' η ΥΠ F ₃ C Connection 247 n^\ / k. 118 mg 27% And connection 248 b F ₃ C S-^ _N ^ Compound 248 From purification of compound 230 using chiral SFC: Lux Cellulose-2 5 µm, 250*30 mm; mobile phase: 55% CO ₂ , 45% MeOH (0.3% iPrNH ₂ ) 114 mg 26%

- 107 045364

Compound 231 o O -( RS F ₃ C From intermediate 11, isoxazole-4-carbaldehyde and 3 M MeMgBr in Et ₂ O 25 mg 19% Compound 233 <PW ^rs N=/ N—, ъ F ₃ C From intermediate 11, pyridine-3-carboxaldehyde and 3 M MeMgBr in Et ₂ O 26 mg 19% Connection 235, And connection 236, *r\_ N^\ / 1 />—(*R ъ F ₃ CS Compound 235 _ *s\..... / 1 />.......<*s i F ₃ C S-\ _N ·^ Compound 236 23 mg 17 mg 8% 6%

- 108 045364

And connection 237, And connection 238 n^\ / 1 />.......<*s ъ AYO F ₃ C Compound 237 ъ aYO F ₃ C Compound 238 From intermediate 11, 1methyl-1 N-pyrazole-4-carbaldehyde and sec- chloride butylmagnesium, 25% by weight, in THF Chiral purification SFC: CHIRALPAK AD-H 5 µm, 250*30 mm; mobile phase: 80% CO ₂ , 20% iPrOH (0.6% iPrNH ₂ ). Compounds 237 and 238 were obtained after additional chiral SFC: CHIRALPAK AD-H 5 µm, 250*30 mm; mobile phase: 80% CO ₂ , 20% iPrOH (0.6% iPrNH ₂ ), then chiral SFC: Chiralcel OD-H 5 µm, 250x21.2 mm; 27 mg 29 mg 9% 10%

- 109 045364

mobile phase: 82% CO ₂ , 18% iPrOH (0.3% iPrNH ₂ ). Connection 284, Ν^\ / I /)—( ^RS 174 mg 50% / \ II J F ₃ C S-^V _N V Connection 284 From intermediate 11, 1methyl-1H-pyrazole-4-carbaldehyde and 2 M n-butylmagnesium chloride in THF 58 mg 20% connection 243 nV\ / 1 />......<*R Ν // \ A Yr! / \ II J F ₃ C Connection 243 And ΝΓ\ / 64 mg 22% YES connection 244 ъ Ar i / \ II J F ₃ C Connection 244 From separation by chiral SFC connection 384: CHIRALPAK IC 5 µm, 250*30 mm; mobile phase: 60%

- 110 045364

CO ₂ , 40% EtOH (0.3% iPrNH ₂ ). Connection 285, N^\ / 1 //—<RS \ P F ₃ C Compound 285 From intermediate I, tert-butyl-4-formyl-1H-pyrazole-1 carboxylate and 2 M iPrMgCl in THF 75 mg 22% connection 245 And ϊΥ......6 ^ΗΝ ^ \| ъ F ₃ C Connection 245 25 mg 7% connection 246 / ъ F ₃ CS Connection 246 From separation by chiral SFC connection 285: CHIRALPAK AD-H 5 µm, 250*30 mm; mobile phase: 70% CO ₂ , 30% EtOH (0.3% iPrNH ₂ ) 24 mg 7%

- 111 045364

Connection 286

Connection 254

From intermediate 11,

CAS3 9021-62-0 and 3 M MeMgBr in Et ₂ O

From separation via chiral

Compound SFC 286: Chiralcel OD-H

CO ₂ , 30% MeOH (0.3% iPrNH ₂ )) methyl-1 H-imide azole-5-carbaldehyde µm, 250x21.2 mm; mobile phase: 70%

- 112 045364

Example B44.

Preparation of compound 195 h ₂ n

Hydrazine monohydrate (34 µl; 0.86 mmol) was added to a solution of intermediate 85 (100 mg; 0.17 mmol) in EtOH (4 ml). The solution was heated at 50°C for 2 hours 30 minutes. The reaction mixture was poured into ice water and extracted with DCM. The organic layer was separated, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 12 g; mobile phase: 90% DCM, 10% MeOH (+10% NH ₄ OH)). The purified fractions were collected and evaporated to dryness. The residue was taken up in Et ₂ O and evaporated to dryness to give 35 mg (45%) of compound 195.

Example B45.

Receiving connection 287 connection 201

- 113 045364 and connections 202

A mixture of intermediate 87 (145 mg; 0.24 mmol) and TFA (0.7 ml; 9.15 mmol) in DCM (7 ml) was stirred at RT. during the night. The reaction mixture was evaporated to dryness. The residue was diluted with DCM and H ₂ O, then increased in basicity with aqueous 3N. NaOH. The organic layer was decanted, dried over MgSO ₄ , filtered and evaporated to dryness to give 100 mg (83%) of compound 287.

Enantiomers were separated using chiral SFC (CHIRALPAK IC 5 µm, 250x20 mm; mobile phase: 50% CO2, 50% MeOH (+2% iPrNH2)). Fractions containing each enantiomer were collected, evaporated to dryness and purified by reverse phase chromatography (YMC-actus Triart-C18 10 µm, 30x150 mm; mobile phase: gradient 75% NH4HCO ₃ 0.2%, 25% ACN to 35% NH4HCO3 0.2%, 65% ACN). Purified fractions were collected, evaporated to dryness and lyophilized from ACN/water (20/80) to yield 21 mg (17%) of compound 201 and 23 mg (19%) of compound 202.

Example B46.

Receiving connection 224

connections 251

- 114 045364 and connections 252

Under N2 atmosphere, a mixture of intermediate 11b (1 g) and 5-acetyl-1-methylpyrazole (168 mg; 1.35 mmol) in THF (15 ml) was stirred at RT. during the night. NaBH(OAc) ₃ (718 mg; 3.4 mmol) was then added portionwise. The reaction mixture was stirred at room temperature for 72 hours, poured into cold water, increased in basicity with K ₂ CO3 powder and extracted with EtOAc. The organic layer was dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 12 g; mobile phase: gradient 100% DCM, 0% MeOH to 95% DCM, 5% MeOH, 0.3% NH4OH). Fractions were collected and evaporated to dryness. The residue (300 mg) was purified a second time by reverse phase chromatography (YMC-actus Triart C18 10 µm, 30x150 mm; mobile phase: gradient from 65% NH4HCO3 0.2% aq, 35% ACN to 25% NH4HCO3 0.2 % aq., 75% ACN). The purified fractions were collected and evaporated to dryness to obtain 104 mg of compound 224.

Compound 224 was subjected to chiral SFC resolution ((stationary phase: Chiralcel

OD-H 5 µm, 250x21.2 mm, mobile phase: 70% CO2, 30% EtOH (0.3% iPrNH ₂ )).

Fractions containing the product were mixed, concentrated and lyophilized (ACN/water: 80/20) to obtain 48 mg of compound 251 and 46 mg of compound 252.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 224, starting from the appropriate starting materials.

Connection number Structure Quantity Exit Compound 232 zyv \=/ Rs\ ъ F ₃ C From intermediate 11b and 4acetylpyridine 63 mg Compound 258 XV/ \=/x xCO F ₃ C From intermediate 11b and 5acetylpyrimidine 45 mg

- 115 045364

Example B47.

Preparation of compound 234 n

A mixture of intermediate 88 (130 mg; 0.225 mmol) and 4 M HCl in 1,4-dioxane (0.7 ml; 2.8 mmol) in MeOH (7 ml) was stirred at RT. in 24 hours. The solution was cooled at 5°C and EtOAc was added. The precipitate was filtered and dried to obtain 111 mg of compound 234 (HCl salt).

Example B48.

Receiving connection 270

and connections 271

A mixture of intermediate 11 (300 mg; 0.854 mmol) and (R)-styrene oxide (293 μl; 2.563 mmol) in EtOH (6 ml) was stirred at 60°C for 4 hours. The reaction mixture was evaporated to dryness and the residue was purified by chromatography on silica gel (SiOH irregular particle, 12 g; mobile phase: gradient from 0% NH ₄ OH, 0% MeOH, 100% DCM to 1% NH ₄ OH, 10% MeOH, 90% DCM). Fractions containing the products were collected and evaporated to dryness to obtain 157 mg of a residue representing the intermediate, which was purified by reverse phase chromatography (YMC-actus Triart C18 10 µm, 30x150 mm; mobile phase: gradient from 60% NH ₄ HcO ₃ 0.2% aq., 40% ACN to 40% NH ₄ HCO ₃ 0.2% aq., 60% ACN). Fractions containing the products were collected, evaporated to dryness and lyophilized from water/ACN (80/20; 10 ml) to obtain 58 mg (15%) of compound 270 and 62 mg (16%) of compound 271.

- 116 045364

Example B50

Receiving connection 105

N

TFA (2.5 ml) was added to a solution of intermediate 93 (500 mg; 0.95 mmol) in DCM (25 ml) and the reaction mixture was stirred for 18 hours. The reaction mixture was poured into 10% aqueous K ₂ CO ₃ solution and extracted with DCM. The organic layer was separated, filtered through Chromabond® and evaporated to dryness. The residue was purified by silica gel chromatography (SiOH irregular particle, 10 g; mobile phase: 0.7% NH ₄ OH, 7% MeOH, 93% DCM). The purified fractions were collected and evaporated to dryness. The residue was crystallized from diisopropylethyl ether and dried to give 120 mg (29%) of compound 105.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 105, starting from the corresponding starting materials.

Connection or number Structure Quantity Exit Connection 110 Js b .-about From intermediate 96 78 mg 34% Connection 109 p in 1 / Ί ⁿ F ₃ C From intermediate compound 98 117 mg 41% Connection 111 if 47 mg 15%

- 117 045364

Example B51.

Receiving connection 89

n. HCl in 1,4-dioxane (0.708 ml; 2.833 mmol) was added to a solution of intermediate 101 (150 mg; 0.283 mmol) in ACN (7.5 ml) and the reaction mixture was stirred at room temperature overnight. The reaction mixture was poured into 10% aqueous K2CO3 solution and extracted with DCM. The organic layer was decanted, filtered through Chromabond® and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 12 g; mobile phase: gradient 0% MeOH, 100% DCM to 15% MeOH, 85% DCM). The purified fractions were collected and evaporated to dryness. The residue was taken up several times with Et ₂ O to give 30 mg (25%) of compound 89.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 89, starting from the corresponding starting materials.

- 118 045364

Connection number Structure Quantity Exit Connection 87 % Ν F ₃ CS From intermediate 102 47 mg 29% Connection 86 /\__, о 1__/s\ N—, ъ neo F ₃ C From intermediate compound 102 55 mg 42% Connection 311 HN / \ \__/ N— F ₃ CS From intermediate 107 181 mg 58%

Example B56.

Receiving connection 317

Intermediate I (266 mg, 0.812 mmol), intermediate 110 (377 mg, 1.62 mmol) and K ₂ CO ₃ in ACN (8 ml) were stirred overnight at 90°C. The mixture was poured into water, then extracted with EtOAc. The organic layer was dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (stationary phase: SiOH irregular particles, 15-40 µm, 40 g, mobile phase: DCM/MeOH: gradient 100/0 to 92/8). The residue was purified by reverse phase silica gel chromatography (stationary phase: YMC-actus Triart-C18 10 µm, 30x150 mm, mobile phase: 0.2% NH ₄ HCO3/ACN: gradient 60/40 to 0/100). Fractions containing the product were collected and evaporated to dryness. The resulting precipitate was solubilized in ACN and 2 eq. was added. 4 n. solution of HC1 in dioxane. The mixture was concentrated and then lyophilized with 20/80 acetonitrile/water to give 0.091 g of compound 317 (HC1 salt).

- 119045364

Example B57

Receiving connection 320

To a solution of intermediate 11b (200 mg) in ethanol (5 ml) was added 1-(3 methoxyphenyl)propan-2-one (200 mg, 1.218 mmol), PtO ₂ (20 mg) and AcOH (2 drops). After stirring at 60°C overnight under H2, the reaction mixture was concentrated to give a residue which was purified by preparative HPLC (column: SunFire, 19x250 mm, 10 µm, mobile phase A: 0.1% TFA/H2O, B: ACN ) to yield 35 mg (12%) of compound 320 as a yellow solid.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 320, starting from the corresponding starting materials.

- 120 045364

- 121 045364

From intermediate 11b and 2fluorophenylacetone Connection 323 BUT — N— / > , TFA salt From intermediate 11b and 4hydroxyphenylacetone 36 mg Connection 324 — N—. r.^xb /F F, TFA salt From intermediate 11b and 1(p-tolyl)propan-2-one 40 mg

Example B57.

Compound 61 was synthesized together with intermediates 68, 68a and 68b. See synthesis protocol for intermediates 68, 68a and 68b.

- 122 045364

Transformation.

Transformation C1.

Receiving connection 196

Solution of compound 273 (106 mg; 0.24 mmol), 37% w/w. aqueous formaldehyde (110 μl; 1.48 mmol) and MgSO ₄ (580 mg; 4.83 mmol) in DCM (5 ml) were stirred at room temperature. within 1 hour. NaBH(OAc) ₃ (614 mg; 2.9 mmol) was added and the reaction mixture was stirred at RT. within 15 hours. The solution was poured into ice water, increased in basicity with K ₂ CO ₃ and extracted with DCM (x2). The organic layer was washed with brine, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by chromatography on silica gel (SiOH irregular particle, 10 g, mobile phase: gradient from 100% DCM, 0% MeOH, 0% NH ₄ OH to 90% DCM, 10% MeOH, 0.5% NH ₄ OH ). The purified fractions were collected, evaporated to dryness and the residue lyophilized from ACN/water (20/80) to give 70 mg (64%) of compound 196.

The compounds shown in the table below were prepared by using a method similar to that described for the preparation of compound 196, starting from the appropriate starting materials.

Connection number Structure Quantity Exit Connection 304a 5-+- Li F ₃ C From connection 269a 50 mg 28% Connection 304b __/ F )—— / s* \/ σ ГШ F ₃ C 52 mg thirty% From connection 269b

- 123 045364

Transformation C2.

Receiving connection 216

A solution of compound 200 (110 mg; 0.25 mmol) and 37% w/w. an aqueous solution of formaldehyde (19 μl; 0.25 mmol) in MeOH (5 ml) was stirred at room temperature. within 3 hours. NaBH4 (19 mg; 0.5 mmol) was added and the reaction mixture was stirred at room temperature. for 15 hours, poured into ice water, rebasified with K ₂ CO ₃ and extracted with DCM (x2). The organic layer was washed with brine, then dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 300 g; mobile phase: gradient 0.1% NH4OH, 5% MeOH, 95% DCM to 1% NH4OH, 10% MeOH, 90% DCM). Purified fractions were collected, evaporated to dryness and lyophilized from 20/80 ACN/water to yield 15 mg (13%) of compound 216.

Transformation C3.

Receiving connection 303

Under N2 atmosphere at 10°C, ethyl bromide (45 μL, 0.6 mmol) was added to a solution of compound 222 (150 mg; 0.34 mmol) and DIPEA (207 μL; 1.2 mmol) in THF (3 ml). The solution was stirred at room temperature. overnight, then poured into chilled water. The product was extracted with EtOAc. The organic layer was dried over MgSO ₄ , filtered and evaporated to dryness. The residue (164 mg) was purified by silica gel chromatography (stationary phase: unmodified irregular silica, 12 g, mobile phase: gradient 100% DCM, 0% MeOH (+10% NH4OH) to 90% DCM, 10% MeOH (+10% NH4OH)). Fractions containing the product were mixed and concentrated to obtain 77 mg of the intermediate fraction, which was further purified using reverse phase (stationary phase: YMC-actus Triart C18 10 µm, 30x150 mm, mobile phase: gradient from 65% NH4HCO3 0.2 %, 35% ACN to 25% NH4HCO3 0.2%, 75% ACN). Fractions containing the product were mixed and concentrated to yield 40 mg of residue, which was lyophilized with 20/80 acetonitrile/water to yield 34 mg (21%) of compound 303 as a white powder.

- 124 045364

Transformation C4.

Receiving connection 316

A solution of compound 315 (114 mg; 0.237 mmol), 37% formaldehyde in water (106 μl; 1.42 mmol) and MgSO ₄ (568 mg) in DCM (5 ml) was stirred at RT. within 1 hour. NaBH(OAc) ₃ (602 mg; 2.84 mmol) was then added and the mixture was stirred at RT. during the night. The reaction mixture was diluted with DCM and increased in basicity with 10% aqueous K2CO3. The organic layer was decanted, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was purified by silica gel chromatography (irregular SiOH, 12 g; mobile phase: gradient 0% NH4OH, 0% MeOH, 100% DCM to 1% NH4OH, 10% MeOH, 90% DCM). Fractions containing product were collected, evaporated to dryness and lyophilized from water/ACN (80/20; 10 ml) to yield 110 mg (94%) of compound 316.

Analytical part.

LCMS (liquid chromatography/mass spectrometry).

General procedure.

High performance liquid chromatography (HPLC) measurements were performed using an LC pump, diode array detector (DAD) or UV detector and column as described in the respective methods. If necessary, additional detectors were included (see table of methods below).

The column flow was directed to a mass spectrometer (MS), which was equipped with an atmospheric pressure ionization source. It is within the purview of one skilled in the art to set custom parameters (eg, scan range, dwell time, etc.) to obtain ions that allow the nominal monoisotopic molecular weight (MW) of a compound to be determined. Data collection was carried out using appropriate software.

Compounds were described by their experimental retention time (Rt) and ion values. Unless otherwise indicated in the data table, the molecular ion indicated corresponds to [M+H] (protonated molecule) and/or [MH] ^- (deprotonated molecule). If the compound was not directly capable of ionization, the type of adduct was indicated (i.e. [M+NH4] ⁺ , [M+HCOO] ^- , etc.). For molecules with complex isotopic distributions (Br, Cl, etc.), the indicated value is the value obtained for the smallest isotope mass. All results are subject to experimental errors, which are usually associated with the method used.

Hereinafter in this document SQD means single quadrupole detector, c.t. means room temperature, BEH means bridged ethyl siloxane/silica, HSS means high strength silica, DAD means diode array detector. All other abbreviations used are previously defined.

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Table 1a

LCMS method codes (flow rate expressed in 1 ml/min;

column temperature (T) in °C; analysis time in minutes)

Code method A Device Column Mobile phase Gradient Flow rate T speakers Analysis time From 90% A in 1 Agilent: 1200 - DAD and MSD6110 Phenomen ex: LunaC18 (5 µm, 2x50 mm) A: 0.1% CF3COOH in water, B: 0.05% CF3COOH B CH3CN for 0.8 min. up to 20% A in 3.7 minutes, holding for 3 minutes, back to 90% A in 2 minutes. 0.8 50 10 Agilent: Phenomen A: 0.1% From 100% A to 0.8 2 1200 - ex: Luna- CF3COOH for 1 min. — 10 DAD and C18 in water, B: up to 40% A for 50

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MSD6110 (5 µm, 2x50 mm) 0.05% CF ₃ COOH in CH ₃ CN 4 min., up to 15% A in 2.5 min., back to 100% A in 2 minutes. 3 Waters: Acquity UPLC® - DAD and Quattro Micro™ Waters: VEN C18 (1.7 µm, 2.1 x 100 mm) A: 95% CH3COON H ₄ , 7 mM/5% CH ₃ CN, B: CH ₃ CN From 84.2% A for 0.49 min. before 10.5% A for 2.18 min., hold for 1.94 min., back to 84.2% A in 0.73 min., hold for 0.73 min. 0.343 40 6.2 4 Waters:Acquity® H-Class DAD and SQD2™ Waters: VEN C18 (1.7 µm, 2.1 x 100 mm) A: 95% CH3COON H ₄ , 7 mM/5% CH ₃ CN, B: CH ₃ CN From 84.2% A to 10.5% A in 2.18 minutes, holding for 1.96 minutes, back to 84.2% A in 0.73 min., hold for 0.73 min. 0.343 40 6.1 5 Shimadzu: LC- MS2020 - SunFire C18, 5 microns, A: 0.1% HCOOH in water, V: From 70% A for 0.4 min. up to 5% A for 2.0 2.6

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SPD- M20A and Alltech 3300 ELSD 50 * 4.6 m M 0.1% HCOOH in _CH3CN 1.2 min., up to 1% A in 1.0 min. 40 6 Waters UPLCQDa PDAdetector ACQUIT YUPLC BEHC18, 1.7 micron, 2.1 * 50 m M A: 0.1% HCOOH in water, B: 0.1% HCOOH in CH ₃ CN From 90% A for 0.1 min. up to 5% A in 1.1 min., holding at 5% A for 0.8 min. 0.6 50 2.0 7 Waters UPLCQDa PDA- detector ACQUITY UPLC BEH C18, 1.7 microns, 2.1 * 100 m m A: 0.1% HCOOH in water, B: 0.1% HCOOH in CH ₃ CN From 80% A for 1.0 min. up to 5% A for 7.0 min., holding at 5% A for 1.0 min. 0.4 50 9.0

Table 1b

LCMS data. Connection no. means connection number; Rt means retention time in min.

No. with unit Rt (min.) [M+N] ⁺ Adduct/ [M-N] Way LCMS 1 2.725 419.2 - 1 2 2.88 425.1 483.4 [M+CH3COO]’ 3 3 2.9 447.2 - 1 4 2.26 421.1 479.4 [M+CH3COO]' 3 5 3.314 459 - 2

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6 3.176 459 - 2 7 3.129 459 - 2 8 3.71 454.9 - 2 9 3.333 421 - 2 10 3,269 421 - 2 eleven 3.652 454.9 - 2 12 3,409 421 - 2 13 2,498 385 - 1 14 2.76 405 463.4 [M+CH3COO]' 3 15 3.516 455 - 2 16 2.88 419.5 463.3 [M+CH3COO]' 4 17 3,074 482.9 - 1 18 2.94 447 - 1 19 3.92 461.2 519.7 [M+CH3COO]' 3 20a 3.15 433.1 491.4 [M+CH3COO]' 3 20b 3.15 433.1 491.4 [M+CH3COO]' 3 21a 3.66 447.4 445.4 3 21b 3.65 447.3 445.3 3 22 3.104 437 - 1 23 2.11 423.5 481.3 [M+CH3COO]' 4 24 2.46 452.5 510.3 [M+CH3COO]' 4 25 2.43 - 466.3 [M+CH3COO]' 3 26 3.28 420.1 - 3 28 2.08 409.1 - 3 29 2.09 409.1 - 3 thirty 2.17 423.2 481.4 [M+CH3COO]' 3

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31 2.67 461 519.3 [M+CH3COO] 3 34 2.16 409.1 467.4 [M+CH3COO]' 3 35 2.26 423.2 481.4 [M+CH3COO]' 3 36 2.39 451.2 509.5 [M+CH3COO]' 3 37 2.375 492 - 1 40 1.98 395.1 - 3 41 1.95 395.4 - 4 44 2.31 371.5 429.2 [M+CH3COO]' 4 45 2,936 433 - 1 46 2,871 433 - 1 47 3.05 463 - 1 48 2.79 545.9 - 1 49 2,901 433 - 1 50 3,019 447 - 1 54 2.66 436.9 - 1 56 2,731 436.9 - 1 57 3,604 449 - 2 58 2.67 385 443.2 [M+CH3COO]' 3 59 2.2 423 481.2 [M+CH3COO]' 3 61 2.11 357 415.1 [M+CH3COO]' 3 62 2.45 495.2 - 3 63 2.55 455.2 513.4 [M+CH3COO]' 3 64 2.92 411 469.1 [M+CH3COO]' 3 65 2.46 413.1 471.2 [M+CH3COO]' 3 66 2.31 427.1 485.2 [M+CH3COO]' 3 67 2.25 441.5 499.4 [M+CH3COO]' 4

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69 2.89 433.1 491.3 [M+CH3COO] 3 71 2.75 509.2 - 3 74 2.54 547.4 605.7 [M+CH3COO]' 3 75 2.56 506.3 564.5 [M+CH3COO]' 3 76 2.23 462.2 520.4 [M+CH3COO]' 3 77 2.01 452.1 520.5 [M+CH3COO]' 3 82 1.95 426.4 484.2 [M+CH3COO]' 4 84 3.05 465.1 523.3 [M+CH3COO]' 3 88 2.13 488.4 546.3 [M+CH3COO]' 3 112 2.24 452.1 510.4 [M+CH3COO]' 3 86 2.20 452.2 510.5 [M+CH3COO-] 3 87 2.13 438.2 496.6[M+CH3COO-] 3 89 2.14 430.1 488.3 [M+CH3COO-] 3 90 2.32 448.1 506.4[M+CH ₃ COO-] 3 105 1.96 426.4 484.3 [M+CH3COO-] 4 108 2.34 452.2 5Iu,4[M+CH ₃ COO-] 3 109 2.13 438.1 496.4[M+CH ₃ COO-] 3 110 1.93 442.5 500.2[M+CH ₃ COO-] 4 111 1.90 442.5 500.3 [M+CH3COO-] 4 163 2.42 489.1 547.4[M+CH ₃ COO-] 3 164 2.43 489.2 547.4[M+CH ₃ COO-] 3 195 1.90 452.5 510.3 [M+CH3COO-] 4 196 2.35 454.2 512.4[M+CH ₃ COO-] 3 197 2.29 437.2 495.5[M+CH ₃ COO-] 3 198 2.25 437.2 495.4[M+CH ₃ COO-] 3

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199 2.25 437.2 495.5[M+CH ₃ COO-] 3 200 2.28 440.1 498.3 [M+CH3COO-] 3 201 1.92 506.5 564.4[M+CH ₃ COO-] 4 202 1.89 506.5 564.5[M+CH ₃ COO-] 4 203 2.70 465.2 523.4[M+CH ₃ COO-] 3 211 2.40 454.1 512.3 [M+CH3COO-] 3 213 2.38 454.1 512.3 [M+CH3COO-] 3 214 2.98 493.3 551.5 [M+CH3COO-] 3 220 2.43 451.1 509.3 [M+CH3COO-] 3 221 2.44 451.1 509.3 [M+CH3COO-] 3 222 2.31 440.1 498.3 [M+CH3COO-] 3 223 2.20 423.0 421.1 3 224 2.51 437.1 495.3 [M+CH3COO-] 3 228 2.68 440.0 498.2[M+CH ₃ COO-] 3 230 2.70 479.2 537.4[M+CH ₃ COO-] 3 231 2.59 424.1 482.3 [M+CH3COO-] 3 232 2.58 434.1 492.3 [M+CH3COO-] 3 233 2.56 434.1 492.3 [M+CH3COO-] 3 234 2.03 478.2 536.4[M+CH ₃ COO-] 3 235 2.91 479.2 537.4[M+CH ₃ COO-] 3 236 2.91 479.2 537.4[M+CH ₃ COO-] 3 237 2.89 479.3 537.5[M+CH ₃ COO-] 3 238 2.89 479.2 537.5[M+CH ₃ COO-] 3 243 2.75 479.2 537.4[M+CH ₃ COO-] 3 244 2.75 479.2 537.4[M+CH ₃ COO-] 3 245 2.54 451.1 449.2 3

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246 2.53 451.1 449.2 3 247 2.69 479.2 537.4[M+CH ₃ COO-] 3 248 2.69 479.2 537.4[M+CH ₃ COO-] 3 249 2.59 440.0 498.2[M+CH ₃ COO-] 3 250 2.42 489.1 547.3 [M+CH3COO-] 3 251 2.50 437.1 495.3 [M+CH3COO-] 3 252 2.50 437.1 495.3 [M+CH3COO-] 3 253 2.34 437.1 495.4[M+CH ₃ COO-] 3 254 2.34 437.1 495.3 [M+CH3COO-] 3 258 2.42 435.1 493.2[M+CH ₃ COO-] 3 259 2.16 426.1 484.3 [M+CH3COO-] 3 269 2.34 458.4 516.4[M+CH ₃ COO-] 4 270 2.52 449.4 507.4[M+CH ₃ COO-] 4 271 2.48 449.4 507.4[M+CH ₃ COO-] 4 273 2.29 440.2 498.5[M+CH ₃ COO-] 3 274 2.71 465.2 523.4[M+CH ₃ COO-] 3 133 2.36 463.2 521.5 [M+CH3COO]' 3 137 2.26 463.5 521.4 [M+CH3COO]' 4 138 2.27 463.5 521.4 [M+CH3COO]' 4 145 2.42 441.1 499.4 [M+CH3COO]' 3 147 2.11 440.1 498.2 [M+CH3COO]' 3 146 2.10 440.1 498.2 [M+CH3COO]' 3 148 2.35 468.2 466.4 3 154 2.46 441.1 499.4 [M+CH3COO]' 3 155 2.44 441.1 499.3 [M+CH3COO]' 3 150 2.05 412.0 470.2 [M+CH3COO]'; 410.0 3 193 2.96 469.2 527.5 [M+CH3COO]'; 467.8 3

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162 2.96 469.2 527.5 [M+CH3COO]·; 467.8 3 156 2.96 469.2 527.4 [M+CH3COO]'; 467.2 3 157 2.81 469.2 527.4 [M+CH3COO]'; 467.2 3 158 2.60 427.1 485.3 [M+CH3COO]' 3 159 2.60 427.1 485.3 [M+CH3COO]' 3 161 2.76 479.2 537.4 [M+CH3COO]'; 477.2 3 166 2.76 479.2 537.3 [M+CH3COO]'; 477.2 3 167 2.75 479.2 537.3 [M+CH3COO]'; 477.1 3 172 2.75 479.2 537.4 [M+CH3COO]'; 477.4 3 173 2.76 479.2 537.4 [M+CH3COO]'; 477.3 3 174 2.75 479.2 537.4 [M+CH3COO]'; 477.2 3 168 2.40 468.2 526.4 [M+CH3COO]'; 466.3 3 169 2.39 468.2 526.4 [M+CH3COO]'; 466.3 3 170 3.20 483.1 541.3 [M+CH3COO]' 3 171 2.93 477.2 535.4 [M+CH3COO]' 3 183 3.09 465.1 523.4 [M+CH3COO]' 3 182 3.48 475.2 533.4 [M+CH3COO]' 3 192 2.90 483.5 541.4 [M+CH3COO]' 4 179 3.03 483.2 541.5 [M+CH3COO]' 3 180 3.04 483.3 541.5 [M+CH3COO]' 3 184 2.73 399.4 457.3 [M+CH3COO]’ 4

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288 2.74 399.1 457.2 [M+CH3COO]·; 397.1 3 185 2.48 441.5 499.4 [M+CH3COO]' 4 187 2.42 441.1 499.3 [M+CH3COO]'; 439.4 3 186 2.87 479.2 537.5 [M+CH3COO]'; 477.2 3 188 2.80 479.5 537.4 [M+CH3COO]'; 523.4 [M+HCOO]' 4 189 2.78 479.5 537.4 [M+CH3COO]'; 523.2 [M+HCOO]' 4 190 2.80 479.5 537.4 [M+CH3COO]' 4 191 2.79 479.5 537.4 [M+CH3COO]' 4 289 3.32 447 505 [M+CH3COO]' 3 290 3.33 447 505 [M+CH3COO]' 3 291 3.25 451 509 [M+CH3COO]' 3 292 3.25 451 509 [M+CH3COO]' 3 293 2.78 413 471 [M+CH3COO]' 3 294 2.77 413 471 [M+CH3COO]' 3 295 2.24 437 495 [M+CH3COO]' 3 296 3.07 447 505 [M+CH3COO]' 3 297 3.07 447 505 [M+CH3COO]' 3 298 3,H 465 523 [M+CH3COO]' 3 299 2.25 437 495 [M+CH3COO]' 3 303 2.42 468 526 [M+CH3COO]' 3 304a 2.69 472 530 [M+CH3COO]' 3 304b 2.70 472 530 [M+CH3COO]' 3 307 3.51 493 551 [M+CH3COO]' 3 309 2.92 and 455 513 [M+CH3COO]' 3

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3.02 311 1.99 426 484 [M+CH3COO]' 3 312 3.76 537 595 [M+CH3COO]' 3 313 3.76 537 595 [M+CH3COO]' 3 314 2.90 472 530 [M+CH3COO]' 3 315 2.22 482 540 [M+CH3COO]' 3 316 2.30 496 554 [M+CH3COO]' 3 317 3.14 465 - 3 318 2.57 502 - 3 320 0.893 477.2 - 5 321 1,300 477.4 - 6 322 6,213 465.3 - 7 323 1.203 463.3 - 6 324 0.692 461.2 - 5

SFCMS methods.

General procedure for SFC-MS methods.

SFC measurements were performed using a supercritical liquid chromatography (SFC) analytical system equipped with a binary pump to deliver carbon dioxide (CO2) and modifier, an automatic dispenser, a column oven, a diode array detector equipped with a flow cell for operation under high pressure, withstanding values up to 400 bar. If equipped with a mass spectrometer (MS), the flow from the column was directed to (MS). It is within the purview of one skilled in the art to set custom parameters (eg, scan range, dwell time, etc.) to obtain ions that allow the nominal monoisotopic molecular weight (MW) of a compound to be determined. Data collection was carried out using appropriate software.

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Table 2a

SFC-MS Analytical Methods (flow rate expressed in 1 ml/min; column temperature (T) in °C; analysis time in minutes; back pressure (BPR) in bar; all other abbreviations used in the table below are previously defined) ________________

Method code Column Mobile phase Gradient Flow velocity T count. Analysis time BPR 1 Phenomenex Lux Cellulose-4 column (3 µm, 100 x 4.6 mm) A: CO ₂ B: MeOH (0.3% iPrNH ₂ ) 30% B, holding for 3 minutes. 3.5 3 35 103 2 Phenomenex Lux Cellulose-2 column (3 µm, 100 x 4.6 mm) A: CO ₂ B: MeOH (0.3% iPrNH ₂ ) 25% B, holding for 3 minutes. 3.5 3 35 103 3 Daicel Chiralpak® AD-3 (3 µm, 100 x 4.6 mm) A: CO ₂ B: 50/50 EtOH/iPrOH (0.3% iPrNH ₂ ) 30% B, holding for 3 minutes. 3.5 3 35 103 4 Daicel Chiralpak® AD-3 (3 µm, 100 x 4.6 mm) A: CO ₂ B: EtOH (0.3% iPrNH ₂ ) 25% B, holding for 3 minutes. 3.5 3 35 103 5 Daicel Chiralpak® AD-3 (3 µm, 100 x 4.6 mm) A: CO ₂ B: EtOH (0.3% iPrNH ₂ ) 30% B, holding for 3 minutes. 3.5 3 35 103

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7 Daicel Chiralpak® AD3 (3 mkm, 100 x 4.6 mm) A: CO ₂ B: EtOH 50% B, hold 3.5 4 (0.3% iPrNH ₂ ) ie 4 min. 35 103 8 Daicel Chiralpak® AD3 (3 mkm, 100 x 4.6 mm) A: CO ₂ B: iPrOH (0.3% iPrNH ₂ ) 20% B, holding for 6 minutes. 3.5 35 6 103 9 Daicel Chiralpak® AD3 (3 mkm, 100 x 4.6 mm) A: CO ₂ B: iPrOH (0.3% iPrNH ₂ ) 30% B, holding 3.5 3 ie 3 min. 35 103 10 Daicel Chiralpak® AD3 (3 mkm, 100 x 4.6 mm) A: CO ₂ B: iPrOH (0.3% iPrNH ₂ ) 40% B, holding 3.5 3 ie 3 min. 35 103 AND Daicel Chiralpak® AD-3 (3 mkm, 100 x 4.6 mm) A: CO ₂ B: MeOH (0.3% iPrNH ₂ ) 25% B, holding 3.5 3 ie 3 min. 35 103

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12 Daicel Chiralpak® AD-3 (3 µm, 100 x 4.6 mm) A: CO ₂ B: MeOH (+0.3% iPrNH ₂ ) 30% B, holding for 3 min. 3.5 3 35 103 16 Phenomenex Lux cellulose 2 (3 µm, 100 x 4.6 mm) A: CO ₂ B: MeOH (0.3% iPrNH ₂ ) 40% B, holding for 6 minutes. 3.5 6 35 103 17 Phenomenex Lux cellulose 2 (3 µm, 100 x 4.6 mm) A: CO ₂ B: MeOH (0.3% iPrNH ₂ ) 50% B, holding for 10 minutes. 3.5 10 35 103 18 Daicel Chiralpak® IC3 (3 µm, 100 x 4.6 mm) A: CO ₂ B: EtOH (0.3% iPrNH ₂ ) 40% B, holding for 3 minutes. 3.5 3 35 103 20 Daicel Chiralcel® OD3 (3 µm, 100 x 4.6 mm) A: CO ₂ B: EtOH (0.3% iPrNH ₂ ) 20% B, holding for 3 minutes. 3.5 3 35 103 21 Daicel Chiralcel® OD3 (3 µm, 100 x 4.6 mm) A: CO ₂ B: EtOH (0.3% iPrNH ₂ ) 30% B, holding for 3 minutes. 3.5 3 35 103

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22 Daicel Chiralcel® OD3 (3 µm, 100 x 4.6 mm) A: CO ₂ B: iPrOH (0.3% iPrNH ₂ ) 20% B while holding 3.5 3 ie 3 min. 35 103 23 Daicel Chiralcel® OD3 (3 µm, 100 x 4.6 mm) A: CO ₂ B: MeOH (0.3% iPrNH2) 30% B, holding 3.5 3 ie 3 min. 35 103 24 Daicel Chiralcel® OJ3 (3 µm, 100 x 4.6 mm) A: CO ₂ B: EtOH (0.3% iPrNH ₂ ) 15% B, holding 3.5 3 ie 3 min. 35 103 25 Daicel Chiralcel® OJ3 (3 µm, 100 x 4.6 mm) A: CO ₂ B: MeOH (0.3% iPrNH ₂ ) 10% B, holding 3.5 3 ie 3 min. 35 103 26 Daicel Chiralcel® OJ3 (3 µm, 100 x 4.6 mm) A: CO ₂ B: MeOH (0.3% iPrNH ₂ ) 30% B, holding 3.5 3 ie 3 min. 35 103 37 Daicel Chiralpak® IC3 (3 µm, 100 x 4.6 mm) A: CO ₂ B: iPrOH (0.3% iPrNH ₂ ) 40% B, holding 3.5 3 ie 3 min. 35 103 38 Daicel Chiralpak® IC3 (3 µm, A: CO ₂ B: MeOH 20% B, holding for 6 minutes. 3.5 6 100 x 4.6 mm) (0.3% iPrNH ₂ ) 35 103 39 Daicel Chiralcel® OD- A: _CO2 15% B, holding 3.5 3 3 (3 µm, 100 x 4.6 mm) B: MeOH (0.3% iPrNH2) ie 3 min. 35 103 40 Phenomenex Lux cellulose 2 A: _CO2 40% B, holding for 3 minutes. 3.5 3 (3 µm, 100 x 4.6 mm) B: EtOH (0.3% iPrNH ₂ ) 35 103

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Table 2b

SFC-MS data (Elution of isomers in order A occurs before elution in order B under the described conditions)

Connection no. R _t (min.) % UV area Elution order of isomers Way SFCMS 20a 2.62 100 A 1 20b 3.00 99.71 IN 1 21a 2.66 98.09 A 2 21b 3.01 96.93 IN 2 82 1.49 99.72 A 5 105 1.65 97.6 IN 5 110 1.32 97.6 A 12

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111 2.27 100 IN 12 163 0.94 100 A 26 164 1.44 100 IN 26 198 1.54 100 A 1 199 2.29 100 IN 1 273 0.75 99.6 A 7 200 1.19 99.6 IN 7 274 1.29 100 A 2 203 2.04 100 IN 2 211 1.58 100 A 12 213 2.22 99.05 IN 12 220 1.29 100 A 18 221 1.90 100 IN 18 235* 2.81 100 A 8 236* 3.46 100 IN 8 237* 1.97 100 A 22 238* 2.43 100 IN 22 243 1.26 100 A 18 244 1.80 99.6 IN 18 245 2.02 100 A 4 246 2.54 99.5 IN 4 247 1.11 99.95 A 16 248 3.20 100 IN 16 251 0.78 100 A 21

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252 1.23 99.95 IN 21 253 0.99 100 A 23 254 1.82 99.2 IN 23 137 0.98 100 A 12 138 1.33 99.81 IN 12 147 1.99 100 IN 5 146 1.46 99.18 A 5 154 1.15 100 A 3 155 1.35 99.14 IN 3 193 1.16 100 A 4 162 1.44 99.04 IN 4 158 1.16 100 A 24 159 1.66 100 IN 24 166 1.07 100 A 9 167 1.31 99.03 IN 9 173 1.22 100 A 20 174 1.81 100 IN 20 168 2.15 99.17 IN 10 169 1.21 100 A 10 179 0.90 100 A AND 180 1.29 99.58 IN AND 184 1.20 100 A 37 288 1.40 98.50 IN 37 185 1.34 100 A 25 187 1.58 100 IN 25 295 1.04 100 A 5 299 1.29 98.11 IN 5

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296 1.40 100 A 9 297 1.88 100 IN 9 290 1.42 100 A 37 289 1.57 100 IN 37 291 3.50 99.92 A 38 292 3.79 100 IN 38 293 1.24 100 A 39 294 1.58 100 IN 39 304a 1.40 100 A 40 304b 1.92 99.02 IN 40

* The order of elution of isomers comp. No. 235 regarding connection. No. 236; conn. No. 237 regarding connection. No. 238.

SFC-MS was also measured for compounds 188, 189, 190 and 191 under the same SFCMS conditions. The results are shown in table. 2c. (Elution of isomers in the order A occurs before B, B occurs before C, C occurs before D.)_________________________________________________

Connection no. Rt (min.) % UV area Elution order of isomers SFCMS method 188 2.78 100 IN 8 189 3.02 96.66 WITH 8 190 2.40 100 A 8 191 3.21 99.15 D 8

Optical rotation (OR).

The amount of optical rotation was measured using a Perkin Elmer 341 polarimeter. Polarized light was passed through the sample at a path length of 1 decimeter and a sample concentration of 0.2-0.4 g per 100 ml. Weigh 2-4 mg of product into a vessel, then dissolve in 1-1.2 ml of spectroscopy solvent (eg DMF). The cuvette was filled with a solution and placed in a polarimeter at a temperature of 20°C. The OR value was read to the nearest 0.004°.

Calculation of concentration: weight in grams x 100 / volume in 1 ml.

Specific rotation (OR): [α] d ²⁰ : (read rotation x 100)/(1,000 dm x concentration).

^d is the sodium D line (589 nm).

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Table 3

OR Data: Wavelength: 589 nm (specifically stated if different); Solvent: DMF (specifically indicated if different);

temperature: 20°C; conc. means concentration (g/100 ml);

________________OR means optical rotation__________________

Connection no. OR(°) Conc. Wavelength(nm) Solvent 20a -31.43 0.28 589 DMF 20b +26.25 0.32 589 DMF 21a -38.48 0.33 589 DMF 21b +42.02 0.326 589 DMF 112 +23.11 0.251 589 DMF 86 -29.55 0.264 589 DMF 87 -26.22 0.267 589 DMF 108 -29.61 0.206 589 DMF 109 +25.37 0.272 589 DMF 198 -28.25 0.308 589 DMF 199 +20.91 0.263 589 DMF 196 +8.92 0.269 589 DMF 200 -11.54 0.26 589 DMF 201 +7.81 0.269 589 DMF 202 -5.38 0.26 589 DMF 203 -6.22 0.225 365 DMF

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211 +5.65 0.248 589 DMF 213 -9.89 0.263 589 DMF 216 -19.57 0.281 589 DMF 220 -22.63 0.243 589 DMF 221 +11.58 0.259 589 DMF 235 -12.02 0.258 589 DMF 236 +8.58 0.233 589 DMF 237 +31.58 0.228 365 DMF 238 -32.09 0.215 365 DMF 243 -30.12 0.332 365 DMF 244 +20.94 0.277 365 DMF 245 -17.12 0.292 589 DMF 247 -17.25 0.255 365 DMF 248 +5.86 0.239 365 DMF 251 -20.6 0.267 589 DMF 252 +12 0.275 589 DMF 253 -25.4 0.252 589 DMF 254 +17.92 0.318 589 DMF 273 +10.38 0.26 589 DMF 274 -5.41 0.222 589 DMF 137 +29.32 0.249 589 DMF 138 -24.9 0.261 589 DMF 147 +8.06 0.273 589 DMF 146 -12.65 0.245 589 DMF 154 -8.59 0.256 589 DMF

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155 +6.64 0.241 589 DMF 150 +14.1 0.234 365 DMF 193 -8.62 0.232 365 DMF 162 -5.45 0.275 589 DMF 158 +16.54 0.26 365 DMF 159 -22.31 0.251 365 DMF 166 -12.6 0.254 589 DMF 167 +7.97 0.276 589 DMF 173 +17.92 0.279 365 DMF 174 -21.99 0.282 365 DMF 168 -4.59 0.283 365 DMF 179 +20.19 0.208 365 DMF 180 -93.02 0.215 365 DMF 184 +17.31 0.219 589 DMF 288 -26.63 0.338 589 MeOH 185 +32.45 0.256 589 DMF 187 -32.45 0.228 589 DMF 289 +36.73 0.245 589 DMF 290 -41.87 0.246 589 DMF 291 -37.05 0.278 589 DMF 292 +32.39 0.247 589 DMF 293 +29.15 0.295 589 DMF 294 -30.83 0.253 589 DMF 295 +27.38 0.263 589 DMF 296 +50.44 0.113 589 DMF 297 -32.45 0.265 589 DMF 299 -33.79 0.29 589 DMF 304a +8.42 0.285 589 DMF 304b -6.49 0.385 589 DMF

NMR.

NMR experiments were performed using a Bruker Avance 500 spectrometer equipped with a 5 mm Bruker BBFO probe head with z-gradients and operating at 500 MHz for protons and 125 MHz for carbon atoms, or using a Bruker Avance DRX 400 spectrometer using an internal deuterium lock and equipped with a reverse double resonance (1H, ¹³ C, SEI) probe head with z-gradients and operating at 400 MHz for protons and 100 MHz for carbon atoms. Chemical shifts (δ) are reported in parts per million (ppm). J values are expressed in Hz.

Alternatively, some NMR experiments were performed using a Bruker Avance III 400 spectrometer at ambient temperature (298.6 K), using an internal deuterium lock, equipped with a 5 mm PABBO BB probe head with z-gradients, and operating at 400 MHz for protons and 100 MHz for carbon atoms. Chemical shifts (δ) are reported in parts per million (ppm). J values are expressed in Hz.

Connection 13.

1H NMR (400 MHz, CHLOROFORM-d) δ ppm 8.42 (s, 1H), 7.38 (s, 1H), 3.95 (br s, 2H), 3.83 (br t, J=6 ,6

Hz, 2H), 3.63 (q, J=10.1 Hz, 2H), 3.27 (br s, 4H), 2.32 (d, J=6.8 Hz, 2H), 2.28 - 2.18 (m, 2H), 1.69 - 1.57 (m,

1H), 0.90 (d, J=6.6 Hz, 6H).

Connection 20b.

^1H NMR (500 MHz, DMSO-d6) δ ppm 11.64 (br s, 1H), 8.49 (s, 1H), 7.77 (s, 1H), 7.67 (br d, J= 3.8 Hz, 2H), 7.35 - 7.51 (m, 3H), 4.32 - 4.48 (m, 2H), 4.03 - 4.18 (m, 2H), 3.91 (br s, 4H), 3.17 - 3.61 (m, 4H), 2.00 2.32 (m, 4H).

Connection 82.

1H NMR (500 MHz, DMSO-d6) δ ppm 8.35 (s, 1H), 7.39 (s, 1H), 4.28 - 4.77 (m, 6H), 4.01 - 4.19 (m, 4H), 3.62 (br t, J=7.9 Hz, 1H), 2.88 (dd, J=9.9, 6.8 Hz, 1H), 2.45 (br dd, J=9.9, 7.7 Hz, 1H), 2.00 - 2.21 (m, 2H), 1.17 - 1.28 (m, 1H), 0.95 (d, J=6, 3 Hz, 3H).

Connection 84.

1H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.33 (s, 1H), 7.72 (s, 1H), 7.22 (dd, J=8.4, 5.8 Hz, 2H) , 7.09 (t, J=9.0 Hz, 2H), 4.08 (q, J=11.0 Hz, 2H), 3.66 - 3.96 (m, 4H), 3.17 ( s, 4H), 2.65 - 2.73 (m, 1H), 2.38 - 2.45 (m, 1H), 2.27 (dd, J=12.9, 8.5 Hz, 1H) , 2.14 (br s, 2H), 0.72 (d, J=6.3 Hz, 3H).

Connection 193.

1H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.32 (s, 1H), 7.70 (s, 1H), 4.07 (q, J=11.1 Hz, 2H), 3.65 - 3.91 (m, 6H), 3.07 - 3.30 (m, 6H), 2.14 (br s, 2H), 1.85 (br s, 1H), 1.70 - 1.82 (m, 1H), 1.63 (br s, 1H), 1.28 - 1.52 (m, 4H), 0.88 (dd, J=9.6, 7.1 Hz, 6H).

Connection 196.

1H NMR (500 MHz, DMSO-d6) δ ppm 8.32 (s, 1H), 7.70 (s, 1H), 4.07 (q, J=10.9 Hz, 2H), 3.52 - 3.97 (m, 4H), 3.34 - 3.43 (m, 2H), 3.02 - 3.22 (m, 4H), 2.72 (dd, J=8.5, 6.3 Hz, 1H), 2.58 (dd, J=8.8, 6.6 Hz, 1H), 2.51 2.53 (m, 1H), 2.28 - 2.35 (m, 1H), 2.19 (dd, J=9.5, 2.2 Hz, 1H), 2.12 (s, 4H), 1.58 (td, J=7.1, 2.2 Hz, 1H), 0 .79 (dd, J=14.8, 6.9 Hz, 6H).

Connection 273.

1H NMR (500 MHz, DMSO-d6) δ ppm 8.32 (s, 1H), 7.67 (s, 1H), 4.03 (q, J=11.0 Hz, 2H), 3.87 ( s, 2H), 3.79 (t, J=6.9 Hz, 2H), 3.61 (br t, J=7.6 Hz, 1H), 3.46 (br t, J=7.9 Hz, 1H), 3.12 - 3.34 (m, 7H), 2.66 - 2.77 (m, 1H), 2.28 (dd, J=9.1, 2.2 Hz, 1H) , 2.15 (t, J=6.9 Hz, 2H), 1.62 (td, J=6.9, 2.2 Hz, 1H), 0.82 (dd, J=14.2, 6 .9 Hz, 6H)

Connection 274

1H NMR (500 MHz, DMSO- _d6 ) δ ppm 8.31 (s, 1H), 7.69 (s, 1H), 7.45 (s, 1H), 7.20 (s, 1H), 4 .07 (q, J=10.9 Hz, 2H), 3.58 - 3.93 (m, 7H), 2.97 - 3.08 (m, 4H), 2.52 - 2.55 (m , 1H), 2.12 (br s, 2H), 1.7 (td, J=6.6, 4.4 Hz, 1H), 0.70 (dd, J=12.6, 6.6 Hz , 6H).

Connection 320.

1HNMR (400 MHz, CD3OD) δ 8.37 (s, 1H), 7.63 (s, 1H), 7.28 (t, J=8.8 Hz, 1H), 6.87-6.86 ( m, 3H), 4.304.18 (m, 5H), 4.08-3.93 (m, 3H), 3.91-3.88 (m, 2H), 3.88-3.85 (s, 3H), 3.72-3.70 (m, 1H), 3.06 (dd, J=5.2 Hz, J=13.6 Hz, 1H), 2.64 (dd, J=9.2 Hz, J=13.6 Hz, 1H), 2.47-2.385 (m, 2H), 1.19-1.18 (d, J=6.8 Hz 3H).

Connection 321.

1H NMR (400 MHz, DMSO) δ 8.38 (d, J=5.2 Hz, 1H), 7.65 (s, 1H), 7.21 (d, J=8.4 Hz, 2H), 6.92 (d, J=8.4 Hz, 2H), 3.75-4.33 (m, 11H), 3.74 (s, 3H), 3.61 (s, 1H), 2.93 (dd, J=3.6 Hz, 12.8 Hz, 1H), 2.27-2.36 (m, 2H), 1.03 (d, J=5.6 Hz, 3H).

Connection 322.

1H NMR (400 MHz, CD3OD) δ 8.40 (s, 1H), 7.66 (s, 1H), 7.38-7.34 (m, 2H), 7.22-1.13 (m, 2H), 4.37-4.26 (m, 5H), 4.13 (s, 1H), 4.01 (s, 2H), 3.91 (q, J=10.4 Hz, 2H), 3.74-3.69 (m, 1H), 2.98 (dd, J=3.6 Hz, 13.2 Hz, 1H), 2.68 (dd, J=9.6 Hz, 13.2 Hz, 1H), 2.47 (s, 2H), 1.20 (d, J=6.4 Hz, 3H).

Connection 323.

1HNMR (400 MHz, CD3OD) δ 8.38 (s, 1H), 7.64 (s, 1H), 7.10 (d, J= 8.4 Hz, 2H), 6.78 (d, J= 8.4 Hz, 2H), 3.86-7.27 (m, 10H), 3.60-6.62 (m, 1H), 2.96 (dd, J=4.8 Hz, 14.0 Hz, 1H), 2.60 (dd, J=4.8 Hz, 14.0 Hz 1H), 2.372.46 (m, 2H), 1.18 (d, J=6.8 Hz, 3H).

Connection 324.

1H NMR (400 MHz, CD3OD) δ 8.36 (s, 1H),7.62 (s, 1H), 7.19-7.15 (m, 4H), 4.28-4.16 (m, 5H), 4.14-3.98 (m, 4H), 3.93-3.87 (m, 2H), 3.66-3.64 (m, 1H), 3.04 (dd, J= 4.8 Hz, J=13.6 Hz, 1H), 2.63 (dd, J=9.2 Hz, J=13.6 Hz 1H), 2.46-2.32 (m, 5H), 1.18 (d, J=6.4 Hz, 3H).

Pharmacological part.

1) Fluorescent polarization analysis of menin-MLL interaction.

To a black 384-well microtiter plate with a non-binding surface, 50 nL of 160X test compound in DMSO and 4 μL of 2X menin in assay buffer (40 mM TrisHCl, pH 7.5, 50 mM NaCl, 1 mM DTT, and 0.001% Tween 20) were added. After incubating the test compound and menin for 10 min at ambient temperature, 4 μl of 2X MBM1 peptide was added.

- 148 045364 labeled FITC (FITC-e-aHTu-SARWRFPARPGT-NH ₂ ) in assay buffer, the microtiter plate was centrifuged at 1000 rpm for 1 min, and the assay mixtures were incubated for 15 min at ambient temperature. The relative amount of menin-FITCMBM1 complex present in the assay mixture was determined by measuring the fluorescence polarization (FP) of the FITC tag using a BMG PHERAstar plate reader (excitation at 485 nm/emission at 520 nm) at ambient temperature. The final reagent concentrations in the binding assay were 100 nM menin, 5 nM FITC-labeled MBM1 peptide, and 0.625% DMSO in assay buffer. Titration procedures for test compounds to investigate their dose-dependent effect were performed using an 11-point, three-fold serial dilution scheme starting at 31 μM.

The potency of compounds was determined by initially calculating the % inhibition for each compound concentration according to the equation 1 % inhibition = ((HC - LC) - (pp ^compound - LC)) / (HC - LC)) * 100 (Eq. 1) where LC and HC represent the FP values in the assay in the presence or absence of a saturating concentration of compound that competes with FITC-MBM1 for binding to menin, and FP compound is the measured FP value in the presence of the test compound. FP HC and LC values are the average of at least 16 replicates per plate. For each test compound, the % inhibition values were plotted against the logarithm of the concentration of the test compound, and the IC ₅₀ value was obtained by fitting these data according to the equation 2 % inhibition = lower + (upper-lower)/(1+10 ^L ( (1o^C5o-1od[com.])*A)) (eq. 2);

where lower and upper represent, respectively, the lower and upper asymptotes of the dose-response curve, IC ₅₀ is the concentration of the compound that produces 50% signal inhibition, and h is the Hill coefficient.

2) Proliferation analysis.

The antiproliferative effect of test compounds inhibiting the menin-MLL protein-protein interaction was assessed in human leukemia cell lines. The MV-4-11 and MOLM14 cell lines contain MLL translocations and express MLL-based fusion proteins MLL-AF4 and MLLAF9, respectively, as well as the wild-type protein from the second allele. Consequently, the MLL rearranged cell lines MV-411 and MOLM14 are characterized by HOXA/MEIS1 gene expression profiles similar to those of stem cells. K562 and KG1 were used as control cell lines containing two wild-type MLL alleles to exclude compounds showing common cytotoxic effects.

MV-4-11 and MOLM14 were cultured in RPMI-1640 (Sigma Aldrich) supplemented with 10% fetal bovine serum (HyClone), 2 mM L-glutamine (Sigma Aldrich), and 50 μg/ml gentamicin (Gibco). K562 was propagated in RPMI-1640 (Sigma Aldrich) supplemented with 20% fetal bovine serum (HyClone), 2 mM L-glutamine (Sigma Aldrich), and 50 μg/ml gentamicin (Gibco). KG1 was cultured in Iscove's MDM (Gibco) supplemented with 20% fetal bovine serum (HyClone), 2 mM L-glutamine (Sigma Aldrich), and 50 μg/ml gentamicin (Gibco). Cells were maintained at 0.3–2.5 million cells per ml during culture, and the number of passages did not exceed 25.

To evaluate antiproliferative effects, 1500 MV-4-11 cells, 300 MOLM14 cells, 750 K562 cells, or 1300 KG1 cells were seeded in 200 μl of medium per well in 96-well round-bottom ultra-low adhesion plates (Costar, catalog number 7007). The number of cells to be seeded was chosen based on growth curves to ensure linear growth throughout the experiment. Test compounds were added at various concentrations and the DMSO content was normalized to 0.3%. Cells were incubated for 8 days at 37°C and 5% CO2. The growth of spheroid-like structures was monitored in real time by live cell imaging (IncuCyteZOOM, Essenbio, 4x objective) with one image taken every four hours for 8 days. Confluence (%) as an indicator of spheroid size was determined using a comprehensive analysis tool.

To determine the cumulative effect of test compounds over time, the area under the curve (AUC) was calculated from a confluency versus time plot. Confluence at the beginning of the experiment (t = 0) was used as the baseline for calculating AUC.

Absolute _IC50 values were calculated according to the following procedure:

% of control = (AUC of sample/AUC of control) x 100,

Control AUC = average AUC of control samples (cells without compound/with DMSO as solvent control).

Fitting was applied to the plot of % of control versus compound concentration.

- 149 045364 nonlinear curve using (ordinary) least squares for fitting. Based on this, the absolute IC ₅₀ value (the half-maximal inhibitory concentration of the test compound causing a 50% antiproliferative effect relative to the solvent control) was calculated.

3) Analysis of menin-MLL interaction by time-resolved homogeneous fluorescence (HTRF).

To an untreated white 384-well microtiter plate, add 40 nL of 200X test compound in DMSO and 4 µL of 2X terbium chelate-labeled menin (see preparation below) in assay buffer (40 mM Tris-HCl, pH 7.5, 50 mM NaCl , 1 mM DTT and 0.05% Pluronic F-127). After incubating the test compound and terbium chelate-labeled menin for 5 min at ambient temperature, 4 µl of 2X FITC-labeled MBM1 peptide (FITC-in-alanineSARWRFPARPGT-NH2) in assay buffer was added and the microtiter plate was centrifuged at 1000 rpm for 1 min, and the test mixtures were incubated for 15 min at ambient temperature. The relative amount of menin FITC-MBM1 complex present in the assay mixture was determined by measuring homogeneous time-resolved fluorescence (HTRF) for the donor-acceptor pair of terbium/FITC fluorophores using a BMG PHERAstar plate reader (excitation at 337 nm/terbium emission at 490 nm/FITC emission at 520 nm) at ambient temperature. The degree of fluorescence resonance energy transfer (HTRF value) is expressed as the ratio of the fluorescence emission intensity values for FITC and terbium fluorophores ( ^Fem 520 nm/P™ 490 nm). The final reagent concentrations in the binding assay were 100 pM terbium chelate-labeled menin, 75 nM FITC-labeled MBM1 peptide, and 0.5% DMSO in assay buffer. Titration procedures for test compounds to investigate their dose-dependent effect were performed using an 11-point, three-fold serial dilution scheme starting at 31 μM.

The potency of compounds was determined by initially calculating the % inhibition for each compound concentration according to Equation 1:

% inhibition = ((HC - LC) - (HTRF ^coe№ " - LC)) / (HC - LC)) * 100 (eq. 7);

where LC and HC are the HTRF values in the assay in the presence or absence of a saturating concentration of the compound that competes with FITC-MBM1 for binding to menin, and HTRF ^o e ^c dine is the measured HTRF value in the presence of the test compound. HTRF HC and LC values are the average of at least 16 replicates per plate. For each test compound, the % inhibition values were plotted against the logarithm of the concentration of the test compound, and the IC ₅₀ value was obtained by fitting these data according to the equation 2 % inhibition = lower + (upper-lower)/(l+10 ^Λ ( (log/C5o-log[coed.])*/2)) (eq. 2);

where lower and upper represent, respectively, the lower and upper asymptotes of the dose-response curve, IC ₅₀ is the concentration of the compound that produces 50% signal inhibition, and h is the Hill coefficient.

Preparation of menin labeled with terbium cryptate. Menin (amino acids 1-610-6xHis tag) was labeled with terbium cryptate as follows. The buffer for 2 mg menin was replaced with 1x phosphate-buffered saline. Incubate 16 μM menin with terbium NHS-cryptate (Cisbio Bioassays, Bedford, MA) in 4-fold molar excess for 2 hours at room temperature. The labeled protein was purified from free label by running the reaction mixture through a Superdex 200 Increase 10/300 GL column at 0.75 ml/min. Peak fractions were collected, aliquoted, and frozen at 80°C.

- 150 045364

Menin protein sequence (SEQ IC NO: 1)

MGLKAAQKTLFPLRSroDVVRLFAAELGREEPDLVLLSLVLGFVEHFLAVNRVI PTNVPELTFQPSPAPDPPGGLTYFPVADLSIIALYARFTAQIRGAVDLSLYPRE GGVSSRELVKKVSDVIWNSLSRSYFKDRAHIQSLFSFITGTKLDSSGVAFAVVG ACQALGLRDVHLALSEDHAWVVFGPNGEQTAEVTWHGKG NEDRRGQTVNA GVAERSWLYLKGSYMRCDRKMEVAFMVCAINPSIDLHTDSLELLQLQQKLLW LLYDLGHLERYPMALGNLADLEELEPTPGRPDPLTLYHKGIASAKTYYRDEHI YPYMYLAGYHCRNRNVREALQAWADTATVIQDYNYCREDEEIYKEFFEVAN DVIPNLLKEAASLLEAGEERPGEQSQGTQSQGSAL QDPECFAHLLRFYDGICKW EEGSPTPVLHVGWATFLVQSLGRFEGQVRQKVRIVSREAEAEAEEPWGEEAR EGRRRGPRRESKPEEPPPPKKPALDKGLGTGQGAVSGPPRKPPGTVAGTARGPE GGSTAQVPAPAASPPPEGPVLTFQSEKMKGMKELLVATKINSSAIKLQLTAQSQ VQMKKQKVSTPSDY TLSFLKRQRKGLHHHHHH

- 151 045364

Table 4

Biological data from menin fluorescence polarization (FP) assay (1), menin-MLL interaction assay by homogeneous time-resolved fluorescence (HTRF) assay (3), and proliferation assay (2). Connection no. means connection number. Values in table. 4 represent average values for all measurements

Connection no. (1) Menin FP assay (1C ₅₀ (µM)) (3) Menin HTRF assay (1C ₅₀ (nM)) (2) Analysis of MV-4-11 spheroids (1C ₅₀ (µM)) (2) Analysis of MOLM14 spheroids (1C ₅₀ (µM)) (2) Analysis of K562 spheroids (1C ₅₀ (µM)) (2) Analysis of KG1 spheroids (1C ₅₀ (µM)) 18 0.14 227 3.9 7.7 >15 1 0.056 144 3.7 4.6 >15 3 0.10 216 3.8 5.6 ~15 14 oh and 178 2.2 4.9 20a 0.076 13 0.96 5 >15 21a 0.49 745 6.9 12.1 >15 21b 0.13 389 4.2 4.6 >15 19 0.81 1960 5 0.67 600 6.4 30.6 >15 12 0.63 2247 >15 >15 >15 13 0.042 39 0.73 4.2 >15 AND 0.76 2374 10 2.31 9 0.84 1579 8 0.79 2525 6 0.24 339 2.4 10.1 >15 7 0.73 1216 4 0.73 3164 2 0.39 706 4.9 14.6 >15 15 0.57 471 14.0 >15 >15 22 0.069 12 0.55 2.0 >15 23 0.95 1208 24 0.49 1486 9.1 >15

- 152 045364

25 0.31 1465 3.8 7.2 26 0.16 143 10.4 >15 197 0.30 541 7.3 >15 86 0.092 102 2.6 11.3 >15 133 0.12 385 2.1 >15 thirty 0.53 839 9.2 >15 88 0.32 486 >15 >15 89 0.045 31 1.3 10.5 90 ο,ιι 288 11.1 >15 74 0.12 211 2.0 6.1 75 2.36 34 0.72 902 198 0.14 178 5.1 12.4 199 0.41 1196 12.9 >15 35 0.99 2375 36 0.4 777 37 0.17 177 1.5 3.1 76 0.68 901 40 0.84 1465 195 1.07 1536 41 0.63 950 273 0.021 8 0.08 0.86 10.3 12 200 ο,ιι 190 3.4 10.3

- 153 045364

201 1075 12.7 >15 202 490 >15 137 1097 Legal entity >15 44 344 4.1 >15 82 9 0.38 1.8 >15 >15 105 15 0.67 3.4 >15 108 753 >15 109 571 8.1 45 330 3.4 46 448 4.1 47 669 48 483 2.0 49 310 2.1 50 665 110 236 4.0 111 394 9.3 77 191 12.1 54 342 3.7 56 304 4.9 112 1540 274 7 0.36 2.3 >15 >15 203 422 7.2 145 37 1.7 7.3

- 154 045364

146 46 2.4 147 341 8.2 57 179 4.2 8.6 148 5 0.33 211 13 0.68 3.5 12.5 213 149 1.9 214 17 0.76 1.4 >15 150 60 2.0 220 75 0.65 4.7 >15 >15 221 1157 222 4 0.2 223 496 6.1 228 238 58 185 2.7 59 1245 230 44 231 107 232 914 233 782 234 1045 235 23 0.71 236 1041 237 52 1.2

- 155 045364

238 659 154 328 155 69 0.7 1.2 >15 243 249 244 848 245 10 0.32 2 >15 246 284 247 19 0.45 2.1 >15 >15 248 549 61 513 249 912 250 190 251 1660 252 1007 253 323 254 861 156 21 0.79 157 101 158 281 159 1174 62 28 0.64 4 >15 161 187 4.2 258 3083

- 156 045364

193 25 0.65 1.7 >15 >15 162 77 3.1 63 201 196 4 o, s 1.1 >15 >15 163 277 164 1125 259 20 1.3 64 1139 65 65 3.2 166 79 2.4 167 1126 168 3 0.19 0.86 11.9 15 169 176 3.5 170 138 4.3 84 20 0.58 1.5 >15 171 159 3.1 66 26 0.95 0.69 >15 172 124 2.9 173 96 3.2 174 1386 67 108 4.9 179 95 1.3 180 130 1.9

- 157 045364

69 186 1.2 182 61 0.78 1.9 >15 183 103 1.3 71 48 1.1 184 73 0.97 >15 186 91 1.3 187 72 1.4 269 8 0.67 4.6 >15 188 115 1.5 189 108 0.92 >15 270 499 271 623 190 524 191 1263 192 619 312 339 313 159 3.6 315 21 0.32 1.5 >15 316 20 θ,4 0.96 >3.7 288 68 1.4 289 6 0.21 1.3 290 25 0.47 1.2 291 13 0.66 1.2 >15

- 158 045364

292 12 0.29 1.5 >15 294 82 1.83 293 81 1.67 307 eleven 0.28 1.5 10.7 299 57 3.18 295 34 1.16 6.7 303 6 0.32 1.02 296 200 4.89 309 16 0.74 1.6 297 242 304b 440 304a 15 0.44 1.3 >15 317 248 0.77 >15 318 72 1.09 298 122 4.23 314 25 0.73 320 259 1.37 321 210 1.1 322 200 1.7 323 47 0.75 324 143 1.9 31 0.087 116 1.3 9.8 224 2365 185 161 1.3 311 85 2.4

- 159 045364

Table 5 Biological data from menin fluorescence polarization (FP) assay (1), menin-MLL interaction assay by homogeneous time-resolved fluorescence (HTRF) assay (3), and proliferation assay (2). Connection no. means connection number. Values in table 5 represent values for individual measurements (not averaged): if the value was determined more than 1 time, each ________________ value is indicated separately in table. 5_____________________

Connection no. (1) Menin FP assay (1C ₅₀ (µM)) (3) Menin HTRF assay (1C ₅₀ (nM)) (2) Analysis of MV-4-11 spheroids (1C ₅₀ (µM)) (2) Analysis of MOLM14 spheroids (1C ₅₀ (µM)) (2) Analysis of K562 spheroids (1C ₅₀ (µM)) (2) Analysis of KG1 spheroids (1C ₅₀ (µM)) 20b 0.038 7 0.51 >15 2.5 >15 >15 16 0.134 63 4.7 >15 9.3 8.7 >15 17 0.054 14 4.9 0.76 0.84 3.4 2.3 AND 28 0.517 1262 0.5 10.8 10.9 3.2 >15 29 0.346 1438 1.8 10.8 >15 9.9 >15 87 0.048 28 4.1 0.63 0.58 0.63 0.68 0.52 8.0 2.8 4.6 >15 138 62 -15.4 1.0 1.4 7

Claims (17)

1. Compound of formula (I) or a tautomer or stereoisomeric form thereof, where R ¹ represents CF3; R ² is selected from the group consisting of hydrogen and CH3; L ¹ is a 7-10-membered saturated spiroheterobicyclic system containing one or two N atoms, provided that it is linked at N to a thienopyrimidinyl heterocycle; And --L ² -R ³ selected from (a), (b), (c), (d), (f) or (g), where (a) L ² selected from the group consisting of >CR ^4a R ^4b and -CHR ^4a CHR ⁵ -; Where L ² is connected to the nitrogen atom L ¹ ; R ^4a is selected from the group consisting of hydrogen; -C(=O)NR ^7a R ^7b ; C _1-4 alkyl, optionally substituted with a substituent selected from the group consisting of -OR ⁸ and -NR ^9a R ^9b ; and a C-linked 4-7 membered non-aromatic heterocyclyl containing at least one nitrogen atom; R ⁵ is selected from the group consisting of hydrogen; -OR ⁶ and C _1-4 αlkyl; R ^4b is selected from the group consisting of hydrogen and methyl; or R ^4a and R ^4b together with the carbon atom to which they are attached form a C _3-5 cycloalkyl or a C 4-6 membered heterocyclyl containing an oxygen atom; wherein each of R ⁶ , R ^7a , R ^7b , R ⁸ , R ^9a and R ^9b is independently selected from the group consisting of hydrogen and C _2-4 alkyl substituted with a substituent selected from the group consisting of -OR ¹¹ and -NR ^10a R ^10b ; wherein each of R ^10a , R ^10b and R ¹¹ is independently selected from the group consisting of hydrogen and C _1-4 alkyl; And R ³ is selected from the group consisting of Ar, Het ¹ , Het ² and a 7-10 membered saturated spirocarbobicyclic system; or (b) L ² represents >CR ^4c R ^4d where R ^4c and R ^4d represent hydrogen; And - 162 045364 r12> ^e^izb 3 tl ^12C 12a 12b 12 R ³ represents ' where R ^12a , R ^12b and R ^12c represents C _1-6 alkyl; or (c) --L2-R ³ is Cl _-6 alkyl, optionally substituted with one, two or three fluorine substituents; or (d) L ² represents O and R ³ represents -CH ₂ -Ar; or R ^1B “ _z 0 (f) --L2-R ³ represents ' where R ¹⁸ is selected from the group consisting of hydrogen and C _1-4 alkyl; R ^18a is selected from the group consisting of hydrogen and fluorine; R ^18b is selected from the group consisting of fluorine, -OC ₁ . ₄ alkyl and C ₁ . ₄ alkyls, optionally substituted 1, 2 or 3 substituents representing a fluorine atom; or R ^18a and R ^18b are bonded to the same carbon atom and together form C ₃ . ₅ cycloalkyl; or (g) --L ² -R ³ represents ' and where Ar represents phenyl, which may be optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -OR ²⁴ and C ₁ . ₄ alkyl, optionally substituted -OR ²⁶ ; Het ¹ is a monocyclic heteroaryl selected from the group consisting of pyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl, 4- or 5-thiazolyl, isothiazolyl and isoxazolyl; or bicyclic heteroaryl selected from the group consisting of indolyl, imidazopyridinyl; each of which may be optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -CN, -OR ²⁴ and C ₁ . ₄ alkyl, optionally substituted with a substituent selected from the group consisting of -CN, -OR ²⁶ and -NR ^27a R ^27b ; And Het ² is a non-aromatic heterocyclyl optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -CN and C ₁ . ₄ alkyl, optionally substituted -OR ²⁶ ; wherein each of R ²⁴ , R ²⁶ , R ^27a and R ^27b is independently selected from the group consisting of hydrogen; C ₁ . ₄ alkyl and C ₂ . ₄ alkyl substituted -NR ^28a R ^28b ; Where R ^28a and R ^28b represent hydrogen; or a pharmaceutically acceptable salt thereof, - 163 045364, provided that the following compounds and their pharmaceutically acceptable addition salts are excluded: - 164 045364 - 165 045364 - 166 045364 - 167 045364 2. Connection according to claim 1, where: (a) L ² is selected from the group consisting of >CR ^4a R ^4b and -CHR ^4a CHR ⁵ -; Where L ² is connected to the nitrogen atom L ¹ ; R ^4a is selected from the group consisting of hydrogen; -C(=O)NR ^7a R ^7b ; Aralkyl, optionally substituted with a substituent selected from the group consisting of -OR ⁸ and - 168 045364 -NR ^9a R ^9b ; and a C-linked 4-7 membered non-aromatic heterocyclyl containing at least one nitrogen atom; R ⁵ is selected from the group consisting of hydrogen; -OR ⁶ and C ₁ . ₄ alkyl; R ^4b is selected from the group consisting of hydrogen and methyl; or R ^4a and R ^4b together with the carbon atom to which they are attached form C ₃ . ₅ cycloalkyl or C-linked 4-6-membered heterocyclyl containing an oxygen atom; wherein each of ^R6 , ^R7a , ^R7b , ^R8 , ^R9a and ^R9b is independently selected from the group consisting of hydrogen and C2. ₄ alkyl substituted with a substituent selected from the group consisting of -OR ¹¹ and -NR ^10a R ^10b ; wherein each of R ^10a , R ^10b and R ¹¹ is independently selected from the group consisting of hydrogen and C ₁ . ₄ αlkyl; And R ³ is selected from the group consisting of Ar, Het ¹ , Het ² and a 7-10 membered saturated spirocarbobicyclic system; or (b) L ² represents >CR ^4c R ^4d where each of R ^4c and R ^4d represents hydrogen; and _R « _a 3 12 12b 12 R ³ represents / where each of R ^12a , R ^12b and R ^12c represents C ₁ . ₆ alkyl; or (c) --L2-R ³ is C ₁ . ₆ alkyl, optionally substituted with one, two or three substituents representing a fluorine atom; or (d) L ² represents O and R ³ represents -CH2-Ar; or 2 3 ' (f) --L2-R ³ represents ' where R ¹⁸ represents hydrogen; or _ , 2 .· ¹ , (g) --L -R represents ' and where Ar is phenyl, optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen and C1. ₄ alkyl, optionally substituted -OR ²⁶ ; Het ¹ is a monocyclic heteroaryl selected from the group consisting of pyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl, 4- or 5-thiazolyl, isothiazolyl and isoxazolyl; or bicyclic heteroaryl selected from imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl; each of which may be optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -CN and C1. ₄ alkyl, optionally substituted with a substituent selected from the group consisting of -CN, -OR ²⁶ and -NR ^27a R ^27b ; And Het ² is a non-aromatic heterocyclyl selected from azetidinyl, pyrrolidinyl and piperidinyl; wherein each of R ²⁶ , R ^27a and R ^27b is independently selected from the group consisting of hydrogen and C1. ₄ alkyl. 3. Connection according to claim 1 or 2, where R ¹ represents CF3; (a) L ² is >CR ^4a R ^4b ; Where R ^4a is selected from the group consisting of hydrogen; -C(=O)NR ^7a R ^7b ; C1. ₄ alkyl and C-linked 4-7 membered non-aromatic heterocyclyl containing at least one nitrogen atom; And R ^4b is selected from the group consisting of hydrogen and methyl; wherein each of R ^7a and R ^7b is independently selected from the group consisting of hydrogen; C1. ₄ alkyl and C ₂ . ₄ alkyl substituted with a substituent selected from the group consisting of -OR ¹¹ and -NR ^10a R ^10b ; wherein each of R ^10a , R ^10b and R ¹¹ is independently selected from the group consisting of hydrogen and C1. ₄ alkyl; And R ³ is selected from the group consisting of Ar, Het ¹ , Het ² and a 7-10 membered saturated spirocarbobicyclic system; or (b) L ² represents >CR ^4c R ^4d where R ^4c and R ^4d represents hydrogen; and _R 12 _B ίΐ' · R ³ represents ' where R ^12a , R ^12b and R ^12c represents C1. ₆ alkyl; or - 169 045364 (c) --L2-R ³ represents C _1-6 alkyl, optionally substituted with one, two or three substituents representing a fluorine atom; or (d) L ² represents O and R ³ represents -CH ₂ -Ar; or (f) --L2-R ³ represents ' where R ¹⁸ represents hydrogen; or (g) --L ² -R ³ represents ' and where Ar represents phenyl, optionally substituted with a substituent representing a halogen atom; Het ¹ is a monocyclic heteroaryl selected from the group consisting of pyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl and 4- or 5-thiazolyl; or bicyclic heteroaryl selected from imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl; each of which may be optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen and C _1-4 alkyl, optionally substituted by a substituent selected from the group consisting of -CN, -OR ²⁶ and - NR ^27a R ^27b ; And Het ² is a non-aromatic heterocyclyl selected from azetidinyl, pyrrolidinyl and piperidinyl; wherein each of R ²⁶ , R ^27a and R ^27b is independently selected from the group consisting of hydrogen and C _1-4 alkyl. 4. Connection according to any one of claims 1-3, where R ¹ represents CF ₃ ; L ¹ is an N-linked 7-10 membered saturated spiroheterobicyclic system containing one or two N atoms selected from the group consisting of (a), (b), (c), (d), (e), ( f) and ⁽ g ^): where a represents the position of binding to the thienopyrimidinyl heterocycle; (a) L ² is >CH ₂ and R ³ is selected from the group consisting of Ar, Het ¹ and a 7-10 membered saturated spirocarbobicyclic system; or (b) --L ² -R ³ is C _1-6 alkyl, optionally substituted with one, two or three fluorine substituents; and where Ar represents phenyl, optionally substituted with a substituent representing a halogen atom; And Het ¹ is a monocyclic heteroaryl selected from the group consisting of 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl and 4- or 5-thiazolyl; or bicyclic heteroaryl selected from imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl; each of which may be optionally substituted with one or two substituents, each of which is independently selected from the group consisting of halogen and C _1-4 alkyl. 5. Connection according to any one of claims 1-4, where R ¹ represents CF ₃ ; R ² represents hydrogen; L ¹ is an N-linked 7-10 membered saturated spiroheterobicyclic system containing one or two N atoms selected from the group consisting of (a), (b), (c), (d), (e), ( f) and (g) - 170 045364 where a represents the position of binding to the thienopyrimidinyl heterocycle; (a) L ² is >CH ₂ and R ³ is selected from the group consisting of Ar, Het ¹ and a 7-10 membered saturated spirocarbobicyclic system; or (b) --L ² -R ³ is C _1-6 alkyl, optionally substituted with one, two or three fluorine substituents; and where Ar represents phenyl, optionally substituted with a substituent representing a halogen atom; And Het ¹ is a monocyclic heteroaryl selected from the group consisting of 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl and imidazolyl; or bicyclic heteroaryl selected from imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl; each of which may be optionally substituted with one or two substituents, each of which is independently selected from the group consisting of halogen and C _1-4 alkyl. 6. The connection according to claim 1, where R ¹ represents CF ₃ ; R ² is selected from the group consisting of hydrogen and CH ₃ ; L ¹ is a 7-10-membered saturated spiroheterobicyclic system containing one or two N atoms, provided that it is linked at N to a thienopyrimidinyl heterocycle; --L ² -R ³ selected from (a), (b), (d), (e) or (f), where: (a) L ² is selected from the group consisting of >CR ^4a R ^4b and -CHR ^4a CHR ⁵ -; Where L ² is connected to the nitrogen atom L ¹ ; R ^4a is selected from the group consisting of -C(=O) NR ^7a R ^7b ; C _1-4 alkyl, optionally substituted with a substituent selected from the group consisting of -OR ⁸ and -NR ^9a R ^9b ; and a C-linked 4-7 membered non-aromatic heterocyclyl containing at least one nitrogen atom; R ⁵ is selected from the group consisting of hydrogen; -OR ⁶ and C _1-4 alkyl; R ^4b is selected from the group consisting of hydrogen and methyl; or R ^4a and R ^4b together with the carbon atom to which they are attached form a C _3-5 cycloalkyl or a C 4-6 membered heterocyclyl containing an oxygen atom; wherein each of R ⁶ , R ^7a , R ^7b , R ⁸ , R ^9a and R ^9b is independently selected from the group consisting of hydrogen and C _2-4 alkyl substituted with a substituent selected from the group consisting of -OR ¹¹ and -NR ^10a R ^10b ; wherein each of R ^10a , R ^10b and R ¹¹ is independently selected from the group consisting of hydrogen and C _1-4 alkyl; And R ³ is selected from the group consisting of Ar, Het ¹ , Het ² and a 7-10 membered saturated spirocarbobicyclic system; or (b) L ² represents >CR ^4c R ^4d where R ^4c and R ^4d represent hydrogen; And R ¹²⁰ * k ^12c · R ³ represents ' where R ^12a , R ^12b and R ^12c represents C _1-6 alkyl; or (d) L ² represents O and R ³ represents -CH ₂ -Ar; or _r ¹⁸ ?Y \ _Z O (f) --L2-R ³ represents > where R ¹⁸ is selected from the group consisting of hydrogen and C _1-4 alkyl; And R ^18a is selected from the group consisting of hydrogen and fluorine; R ^18b is selected from the group consisting of fluorine, -OC _1-4 alkyl and C _1-4 alkyl, optionally substituted with 1, 2 or 3 substituents representing a fluorine atom; or R ^18a and R ^18b are bonded to the same carbon atom and together form a C _3-5 cycloalkyl; and where Ar represents phenyl, which may be optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -OR ²⁴ and C _1-4 alkyl, optionally substituted -OR ²⁶ ; Het ¹ is a monocyclic heteroaryl selected from the group consisting of pyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl, 4- or 5-thiazolyl, - 171 045364 isothiazolyl and isoxazolyl; or bicyclic heteroaryl selected from the group consisting of indolyl and imidazopyridinyl; each of which may be optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -CN, -OR ²⁴ and C^alkyl, optionally substituted by a substituent selected from the group consisting of -CN, -OR ²⁶ and -NR ^27a R ^27b ; And Het ² is a non-aromatic heterocyclyl, optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -CN and C1. ₄ alkyl, optionally substituted -OR ²⁶ ; wherein each of R ²⁴ , R ²⁶ , R ^27a and R ^27b is independently selected from the group consisting of hydrogen; C1. ₄ alkyl and Cr^alkyl substituted -NR ^28a R ^28b ; Where R and R represents hydrogen. 7. The compound according to claim 1, where -L ² -R ³ represents (a); R ³ represents Het ¹ or Het ² ; Het ¹ is a monocyclic heteroaryl selected from the group consisting of pyridyl, 4-, 5- or 6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl and imidazolyl; each of which may be optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -CN, -OR ²⁴ and C1. ₄ alkyl, optionally substituted with a substituent selected from the group consisting of -CN, -OR ²⁶ and -NR ^27a R ^27b ; And Het ² is a non-aromatic heterocyclyl selected from the group consisting of azetidinyl, pyrrolidinyl and piperidinyl; each of which may be optionally substituted with one, two or three substituents, each independently selected from the group consisting of halogen, -CN and C1. ₄ alkyl, optionally substituted with -OR ²⁶ . 8. A pharmaceutical composition containing a compound according to any one of claims 1 to 7 as an inhibitor of protein-protein interactions of menin-MLL, where MLL is a histone methyltransferase, and a pharmaceutically acceptable carrier or diluent. 9. A method for preparing a pharmaceutical composition according to claim 8, comprising mixing a pharmaceutically acceptable carrier with a therapeutically effective amount of a compound according to any one of claims. 1 -7. 10. Use of a compound according to any one of claims 1 to 7 as a medicinal product for the treatment, prevention, amelioration, control or risk reduction of disorders associated with the interaction of menin with MLL proteins and oncogenic MLL-based fusion proteins. 11. Use of a compound according to any one of claims 1 to 7 for the prevention or treatment of cancer, myelodysplastic syndrome (MDS) and diabetes. 12. Use 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. Use according to claim 12, where the leukemia is selected from types of acute leukemia, types of chronic leukemia, types of myeloid leukemia, types of myelogenous leukemia, types of lymphoblastic leukemia, types of lymphocytic leukemia, types of acute myelogenous leukemia (AML), types of chronic myelogenous leukemia (CML) ), types of acute lymphoblastic leukemia (ALL), types of chronic lymphocytic leukemia (CLL), types of T-cell prolymphocytic leukemia (T-PLL), types of large granular lymphocyte leukemia, hairy cell leukemia (HCL), types of leukemia with MLL rearrangement, types of leukemia , MLL-PTD-associated leukemias, MLL-amplified leukemias, MLL-positive leukemias, and leukemias characterized by HOX/MEIS1 gene expression profiles. 14. A method of treating or preventing a disorder selected from cancer, myelodysplastic syndrome (MDS) and diabetes, comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 7 or a pharmaceutical composition according to claim 8. 15. The method of claim 14, wherein the disorder is cancer. 16. The method of claim 15, 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. 17. The method according to claim 15 or 16, where the leukemia is selected from types of acute leukemia, types of chronic leukemia, types of myeloid leukemia, types of myelogenous leukemia, types of lymphoblastic leukemia, types of lymphocytic leukemia, types of acute myelogenous leukemia (AML), types of chronic myelogenous leukemia (CML), types of acute lymphoblastic leukemia (ALL), types of chronic lymphocytic leukemia (CLL), types of T-cell prolymphocytic leukemia (T-PLL), large granular lymphocyte leukemia, hairy cell leukemia (HCL), types of leukemia with MLL rearrangement, types of leukemia associated with MLL-PTD, types of leukemia with MLL amplification, types of MLL-positive leukemia and types of leukemia characterized by HOX/MEIS1 gene expression profiles.