WO2025210042A1

WO2025210042A1 - Combination of zongertinib with a sos1 inhibitor for use in the treatment of cancer

Info

Publication number: WO2025210042A1
Application number: PCT/EP2025/058892
Authority: WO
Inventors: Robyn Leigh SCHENK; Anke Baum; Riccardo Giovannini; Niklas Helge HEINE; Marco Hans HOFMANN; Christoph HOHN; Elke Langkopf; Stephan Georg Mueller
Original assignee: Boehringer Ingelheim International GmbH
Current assignee: Boehringer Ingelheim International GmbH
Priority date: 2024-04-03
Filing date: 2025-04-02
Publication date: 2025-10-09
Anticipated expiration: 2026-10-03

Abstract

The present invention refers to the combination of the HER2 tyrosine kinase inhibitor zongertinib with SOS1 inhibitors. The combination of the invention is useful for the treatment and/or prevention of oncological and/or hyperproliferative diseases, such as cancer. Accordingly, compounds for use, methods of prevention and/or treatment, uses, pharmaceutical compositions and kits related to the combination are herewith provided.

Description

COMBINATION OF ZONGERTINIB WITH A SOS1 INHIBITOR FOR USE IN THE TREATMENT OF CANCER

FIELD OF THE INVENTION

The present invention refers to the combination of the HER2 tyrosine kinase inhibitor zongertinib, or a pharmaceutically acceptable salt thereof, with a SOS1 (Son of Sevenless) inhibitor. The combination of the invention is useful for the treatment and/or prevention of oncological and/or hyperproliferative diseases, in particular cancer.

BACKGROUND OF THE INVENTION

Human epidermal growth factor receptor 2 (HER2; encoded by ERBB2) belongs to the family of ERBB transmembrane receptor tyrosine kinases.

Aberrant activation of HER2 through different mechanisms plays a crucial role in the development and progression of a variety of cancers. For example, more than 20% of human breast cancers overexpresses HER2 resulting from ERBB2 amplification and manifests a historically poor prognosis (Swain, S. M., Shastry, M. & Hamilton, E. Nat. Rev. Drug Discov. 22, 101-126; 2023). Also, in 12 to 20% of metastatic gastric, gastroesophageal junction or esophageal adenocarcinoma cases, HER2 is overexpressed (Jorgensen JT, Hersom M. J Cancer; 3, p. 137-144; 2012; Wu H, et al. Tumori; 103(3), p. 249-254; 2017).

In addition to overexpression, HER2 can be aberrantly activated by oncogenic mutations in multiple solid cancers. For instance, HER2 mutations, most of them within the exon 20, have emerged as important oncogenic drivers in a subset of non-small cell lung cancers (NSCLC) (Connell, C. M. & Doherty, G. J. Esmo Open 2, e000279 (2017)). This subset of NSCLC, especially those carrying the most common 12 base pair insertion in ERBB2 exon 20 resulting in the duplication of the amino acids YVMA in HER2 (HER2^YVMA), is associated with aggressive disease progression, limited treatment options and poor clinical outcomes.

ERBB signaling can also be aberrantly activated via alterations in the ligands to the receptors of the ERBB family. In this context, fusions of the neuregulin-1 gene (NRG1), although very rare, are well documented.

Small molecule tyrosine kinase inhibitors (TKI) are known.

Zongertinib is a potent and selective tyrosine kinase inhibitor of wild type and mutant HER2 that spares wild type epithelial growth factor receptor (EGFR; also referred to as HER1 and encoded by EGFR), another member of the ERBB family. Zongertinib is described in WO 2021/213800 and represents a promising treatment option for HER2 aberrant cancers.

RAS-family proteins including KRAS (V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog), NRAS (neuroblastoma RAS viral oncogene homolog), HRAS (Harvey murine sarcoma virus oncogene) and MRAS (Muscle RAS viral oncogene homolog) and any mutants thereof are small GTPases that exist in cells in either GTP -bound or GDP-bound states and which have a weak intrinsic GTPase activity and slow nucleotide exchange rates (Moore et al., Nat Rev Drug Discov., 2020 Aug;19(8):533-552). Binding of GTPase activating proteins (GAPs) such as NF1 increases the GTPase activity of RAS- family proteins. The binding of guanine nucleotide exchange factors (GEFs) such as S0S1 (Son of Sevenless 1) promote release of GDP from RAS-family proteins, enabling GTP binding. When in the GTP -bound state, RAS-family proteins are active and engage effector proteins including C-RAF and phosphoinositide 3-kinase (PI3K) to promote the RAF/mitogen or extracellular signal-regulated kinases (MEK/ERK) pathway, PI3K/AKT/mammalian target of rapamycin (mTOR) pathway and RalGDS (Rai guanine nucleotide dissociation stimulator) pathway. These pathways affect diverse cellular processes such as proliferation, survival, metabolism, motility, angiogenesis, immunity and growth (Moore et al., Nat Rev Drug Discov., 2020 Aug;19(8):533-552).

Cancer-associated mutations in RAS-family proteins suppress their intrinsic and GAP -induced GTPase activity leading to an increased population of GTP-bound/active RAS-family proteins. This in turn leads to persistent activation of effector pathways (e.g. MEK/ERK, PI3K/AKT/mTOR, RalGDS pathways) downstream of RAS-family proteins. KRAS mutations (e.g. amino acids G12, G13, Q61, A146) are found in a variety of human cancers including lung cancer, colorectal cancer and pancreatic cancer. Mutations in HRAS (e.g. amino acids G12, G13, Q61) and NRAS (e.g. amino acids G12, G13, Q61, A 146) are also found in a variety of human cancer types however typically at a lower frequency compared to KRAS mutations; Moore et al., Nat Rev Drug Discov., 2020 Aug;19(8):533-552). Alterations (e.g. mutation, over-expression, gene amplification) in RAS-family proteins have also been described as a resistance mechanism against cancer drugs such as the EGFR antibodies cetuximab and panitumumab (Leto et al., J. Mol. Med. (Berl). 2014 Jul;92(7):709-22) and the EGFR tyrosine kinase inhibitor osimertinib/AZD9291 (Eberlein et al., Cancer Res., 2015, 75(12):2489-500). Resistance mechanisms were also described upon treatment with G12Ci (adagrasib, sotorasib), including the enrichment for secondary KRAS mutations as well as other oncogenic alleles (Awad et al, N Engl J Med 2021; 384:2382-239). Published data furthermore indicate S0S1 inhibitors could overcome acquired resistance to KRAS G12C inhibition mediated by KRAS secondary mutations (Koga T. et al., Journal of Thoracic Oncology, 2021, 16, 8, 1321 - 1332).

S0S1 is a multi-domain protein with two binding sites for RAS-family proteins: A catalytic site that binds GDP-bound RAS-family proteins to promote guanine nucleotide exchange and an allosteric site that binds GTP-bound RAS-family proteins, the latter causing further increase in the catalytic GEF function of S0S1. Published data indicate a critical involvement of S0S1 in mutant KRAS activation and oncogenic signaling in cancer (Jeng et al., Nat. Commun., 2012, 3: 1168, Hofmann, Gmachl, Ramharter et al, Cancer Discov. 2021, 11(1): 142-15). Depleting S0S1 levels decreased the proliferation rate and survival of tumor cells carrying a KRAS mutation whereas no effect was observed in KRAS wild type cell lines and the effect of loss of SO SI could not be rescued by introduction of a catalytic site mutated SOS1.

Alterations in SOS1 have been implicated in cancer. SOS1 mutations are found in embryonal rhabdomyosarcoma, Sertoli cell testis tumors, granular cell tumors of the skin (Denayer et al., Genes Chromosomes Cancer, 2010, 49(3):242-52), lung adenocarcinoma (Cancer Genome Atlas Research Network., Nature. 2014, 511(751 l):543-50), bladder cancer (Watanabe et al., IUBMB Life., 2000, 49(4):317-20) and prostate cancer (Timofeeva et al., Int. J. Oncol., 2009, 35(4):751-60). In addition to cancer, hereditary S0S1 mutations are implicated in the pathogenesis of RASopathies like e.g. Noonan syndrome (NS) (Pierre et al., Biochem. Pharmacol., 2011, 82(9): 1049-56).

Selective pharmacological inhibition of the binding of the catalytic site of S0S1 to RAS-family proteins was shown to prevent SOSl-mediated activation of RAS-family proteins to the GTP-bound form (Hofmann, Gmachl, Ramharter et al, Cancer Discov. 2021, 11(1): 142-15). Such S0S1 inhibitor compounds can consequently inhibit signaling in cells downstream of RAS-family proteins (e.g. ERK phosphorylation). In cancer cells associated with dependence on RAS-family proteins (e.g. KRAS mutant cancer cell lines), S0S1 inhibitor compounds can deliver anti-cancer efficacy (e.g. inhibition of proliferation, survival etc.). High potency towards inhibition of SOSLRAS-family protein binding and ERK phosphorylation are therefore desirable characteristics for a S0S1 inhibitor compound, preferably coupled with a good metabolic stability and permeability suitable for oral absorption.

Despite all these approaches, there is still a need for improved treatment options for cancer patients. It is therefore an object of the present invention to provide anti -cancer therapies with therapeutic efficacy and safety.

SUMMARY OF THE INVENTION

In the present invention, it was surprisingly discovered that the use of zongertinib in combination with SOS 1 inhibitors has the potential to improve clinical outcome compared to the use of either agent alone. Further it was found the combination of zongertinib with S0S1 inhibitors inhibits the proliferation of certain cancer cells synergistically, when the SOS1 inhibitor monotherapy has little effect.

The present invention thus provides a combination of zongertinib or a pharmaceutically acceptable salt thereof with a SOS 1 inhibitor. The combination is particularly useful in the treatment and/or prevention of cancer, especially wherein said cancer is HER2 aberrant, including HER2 mutant, HER2 overexpressed and/or HER2 amplified.

According to a first aspect, zongertinib or a pharmaceutically acceptable salt thereof is provided for use in the treatment and/or prevention of cancer, wherein zongertinib or the pharmaceutically acceptable salt thereof is administered in combination with a SOS1 inhibitor. Another aspect relates to a SOS1 inhibitor for use in the treatment and/or prevention of cancer, wherein the SOS1 inhibitor is administered in combination with zongertinib or a pharmaceutically acceptable salt thereof.

Another aspect relates to a method of treating and/or preventing cancer, the method comprising administering to a patient in need thereof: (i) zongertinib or a pharmaceutically acceptable salt thereof, and (ii) a SOS1 inhibitor.

Another aspect relates to a use of zongertinib or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment and/or prevention of cancer, wherein zongertinib or the pharmaceutically acceptable salt thereof is administered in combination with a SOS1 inhibitor.

Another aspect relates to a use of a SOS1 inhibitor for the manufacture of a medicament for the treatment and/or prevention of cancer, wherein the SOS1 inhibitor is administered in combination with zongertinib or a pharmaceutically acceptable salt thereof.

Another aspect relates to a pharmaceutical composition comprising: (i) zongertinib or a pharmaceutically acceptable salt thereof, and (ii) a SOS1 inhibitor.

Another aspect relates to a kit comprising: (i) a first pharmaceutical composition comprising zongertinib or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient; and (ii) a second pharmaceutical composition comprising a SOS1 inhibitor.

The definitions of zongertinib, SOS1 inhibitor, combination therapy and cancer provided in the detailed description hereinbelow are applicable to any and all aspects described in the present summary of invention.

In the present invention, any aspect or embodiment referring to a feature (e.g. the identity of the SOS1 inhibitor) can be combined to any one or more aspect(s) or embodiment(s) referring to (an)other feature(s) (e.g. the identity of the cancer) to provide further aspects or embodiments of the invention.

BRIEF DESCRIPTION OF THE FIGURE

Figure 1: Box and whisker plots of the IC50 for zongertinib (zongertinib mono) or the combination of zongertinib and 1.5 pM SOS1 inhibitor of formula (I) (zongertinib + SOSli combi) in PC-9 YVMA (panel A) or OE-19 (panel B). PC-9_YVMA is a NSCLC cell line with the YVMA HER2 exon 20 insertion mutation. OE-19 is a HER2 amplified esophageal cancer cell line. Each dot represents one experiment where zongertinib, a SOS1 inhibitor (SOSli) of formula (I) and their combination were tested (each dot has a different SOSli). Prior to t-testing, normality was tested using D’Agostino & Pearson’s test. For distributions that significantly differed from the normal distribution, statistical significance was determined by one-tailed paired Wilcoxon matched-pairs signed rank test. For distributions that were normal, statistical significance was determined by one-tailed paired t-testing. **** indicates a p-value less than 0.0001. Each panel reports results of two replicates, labelled as “rep”. DESCRIPTION OF THE INVENTION

Zongertinib

As used herein, zongertinib refers to the compound as defined below or to a pharmaceutically acceptable salt thereof:

Zongertinib

The IUPAC name of zongertinib is N-{ l-[8-({3-methyl-4-[(l-methyl-lH-l,3-benzodiazol-5- yl)oxy]phenyl}amino)-[l,3]diazino[5,4-d]pyrimidin-2-yl]piperidin-4-yl}prop-2-enamide. In case of discrepancy between IUPAC name and depicted formula, the formula shall prevail. WO 2021/213800 discloses zongertinib as example compound 1-01 and provides a procedure for its synthesis. Properties of zongertinib and evidence for inhibitory effect on HER2 wild-type and YVMA kinase activity, while sparing EGFR, are also disclosed in WO 2021/213800, which is herein incorporated by reference.

Zongertinib as used herein also encompasses any tautomers, pharmaceutically acceptable salts, solid state forms of the compound, as well as solvates, including hydrates and solvates of pharmaceutically acceptable salts thereof.

In embodiments, zongertinib is a free base. Therefore, in any aspect or embodiment, the expression “zongertinib or a pharmaceutically acceptable salt thereof’ can be replaced by “zongertinib”, without a reference to the pharmaceutically acceptable salt thereof. In embodiments, pharmaceutically acceptable salts of zongertinib are used. The term “pharmaceutically acceptable” used herein refers to compounds, materials, compositions and/or dosage forms which are, within the scope of sound medical judgement, suitable for use in contact with the tissues of human beings without excessive toxicity, irritation, allergic response, or other problem or complication, and commensurate with a reasonable benefit/risk ratio.

As used herein, “pharmaceutically acceptable salts” of zongertinib refers to zongertinib wherein the compound is modified by making acid or base salts thereof. The term pharmaceutically acceptable salts as used herein generally includes both acid and base addition salts. Pharmaceutically acceptable acid addition salts refer to those salts which retain the biological effectiveness and properties of the free base and which are not biologically or otherwise undesirable, formed with inorganic acids or organic acids. Pharmaceutically acceptable base addition salts include salts derived from inorganic bases or organic nontoxic bases. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. For example, such salts include salts from benzenesulfonic acid, benzoic acid, citric acid, ethane sulfonic acid, fumaric acid, gentisic acid, hydrobromic acid, hydrochloric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, 4-methyl-benzenesulfonic acid, phosphoric acid, salicylic acid, succinic acid, sulfuric acid and tartaric acid. In embodiments, pharmaceutically acceptable salts are selected from chloride and fumarate salts.

Pharmaceutically acceptable salts can be synthesized from zongertinib by conventional chemical methods. Generally, such salts can be prepared by reacting the free base form of zongertinib with a sufficient amount of the appropriate acid or base in water or in an organic diluent or solvent like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile, or a mixture thereof.

The term “solvate” as used herein refers to an association or complex of one or more solvent molecules and zongertinib and/or SOS1 inhibitor. Examples of solvents include water, isopropanol, ethanol, methanol, dimethyl sulfoxide (DMSO), ethyl acetate, acetic acid, tert-butyl methyl ether, tetrahydrofuran, methylethyl ketone, N-methylpyrrolidone and ethanolamine. The term “hydrate” refers to a complex where the solvent molecule is water.

In embodiments, zongertinib is in an amorphous form. In embodiments, zongertinib is in a crystalline form.

SOS1 inhibitor

The term “SOS1 inhibitor” as used herein refers to a compound which binds to SOS1 and thereby prevents the SOS1 mediated nucleotide exchange and subsequently reduces the levels of RAS in its GTP bound form. More specifically, a SOS1 inhibitor compound shows a pharmacological inhibition of the binding of the catalytic site of SOS1 to RAS-family proteins. SOS1 inhibitors belonging to different compound classes are known. The term “SOS1 inhibitor” as used herein also encompasses any tautomers, free forms, pharmaceutically acceptable salts, solid state forms of any SOS1 inhibitor inhibitor, as well as solvates, including hydrates and solvates of pharmaceutically acceptable salts thereof.

Examples of SOS1 inhibitors include those described in: Cancer Discovery (2021) 11 (1): 142-157; Med.Chem.2022, 65, 9678-9690; Proc Natl Acad Sci 2019, 116(7):2551-2560; WO 2019/122129; WO 2018/115380, all of which are herein incorporated by reference.

In a preferred embodiment, the SO SI inhibitor is a compound according to formula (I)

wherein the groups R¹ to R⁴, A¹, A², A³, ring B, V, W, p, q and r have one of the meanings given hereafter; and is particularly suitable for the treatment of pathophysiological processes associated with or modulated by SOS1 inhibition, particularly for the treatment of primary and metastatic tumors associated with dependence on RAS-family protein signaling. Therefore, the compounds of formula (I) or the salts thereof as defined herein are particularly suited for the treatment of cancer associated with dependence on RAS-family protein signaling, including sizeable proportions of NSCLC (non-small cell lung cancer) patients.

The compounds of formula (I) exhibit several advantageous properties, such as high potency shown in vitro by inhibiting the interaction between SOS1 and KRAS alleles G12D and G12C with IC50 values below 300 nM, preferably below 200 nM, more preferably below 100 nM, most preferably below 70 nM (see Table 1). Favorable binding affinity to human S0S1 in combination with favorable cellular activity, as shown by the in vitro ERK phosphorylation assay, and favorable pharmacokinetic properties can enable lower doses for pharmacological efficacy. Lower doses have the advantages of lower "drug load" or "drug burden" (parent drug and metabolites thereof) for the patient causing potentially fewer side effects, and lower production costs for the drug product.

Furthermore, the high cellular potency of the compounds of formula (I) is displayed by IC50 values below 600 nM, preferably below 300 nM, more preferably below 200 nM, most preferably below 100 nM in an in vitro ERK phosphorylation assay (see Table 2). In addition to the affinity assay demonstrating the binding of the compounds of formula (I) to the target, the cellular ERK phosphorylation assays are used to examine the potency with which compounds inhibit the S0S1- mediated signal transduction in a KRAS mutant human cancer cell line. This demonstrates the molecular mode of action of compounds by interfering with the RAS-family protein signal transduction cascade. Low IC50 values are indicative of high potency of the S0S1 inhibitor compounds in this assay setting. It is observed that the compounds of formula (I) demonstrate an inhibitory effect on ERK phosphorylation in a KRAS mutant human cancer cell line, thus confirming the molecular mode of action of the S0S1 inhibitor compounds on RAS-family protein signal transduction.

Further, the compounds of formula (I) are metabolically stable in human hepatocytes (metabolically stable in human hepatocytes in this respect is defined as below or equal to 45 % QH, preferably below or equal to 35 %QH, more preferably below or equal to 25% QH, most preferably below or equal to 20%QH, (see Table 3) and the definition of how to calculate the %QH= hepatic blood flow herein below). Therefore, the compounds of formula (I) are expected to have a favorable in vivo clearance and thus the desired duration of action in humans.

In addition, the compounds of formula (I) are characterized by a low DDI risk based on the cytochrome P450 (CYP) inhibition. The DDI perpetrator risk can be indicated by the reversible inhibition of CYP3A4 isoform, wherein an IC50 >50pM represents a low inhibition (see Table 4). Another aspect of the perpetrator potential can be evaluated by mechanism-based inhibition (MBI) of CYP3A. Further, the compounds of formula (I) show a low risk for mechanism-based inhibition (as defined by the remaining CYP3A activities: preferably above or equal to 75% Ctrl, after a preincubation with 25 pM compound for 30 min. most preferably above or equal to 90% Ctrl, (see Table 5 and the definition of how to calculate the %ctrl. is outlined below).

In summary, the compounds of formula (I) are highly potent inhibitors of the protein-protein interaction between S0S1 and RAS, especially KRAS mutated in position 12 or 13, preferably G12C or G12D mutant KRAS, display high cellular potency as seen in an in vitro ERK phosphorylation assay, are metabolically stable in human hepatocytes and show a low DDI risk.

In one aspect, the S0S1 inhibitor relates to compounds of formula (I) wherein each R¹ is independently selected from the group consisting of Ci- ealkyl, Ci-ehaloalkyl and halogen; p denotes 1, 2 or 3;

R² is H, Me or Et;

V is nitrogen (-N=) or carbon

W is nitrogen (-N=) or carbon at least one of V and W is nitrogen (-N=);

A¹, A² and A³ are each independently selected from nitrogen (-N=) or carbon (=CH- or =CR³-); and ' is a single or double bond; 12-0519-WO-1 e_{ach R3, if present, is independently selected from the group consisting of C1-6alkyl,} C_1-6alkoxy, halogen and C_1-6haloalkyl; q_{denotes 0, 1 or 2;} _{ring system B is selected from C3-10cycloalkyl, C4-10cycloalkenyl, 4-13 membered heterocyclyl,} _{5 r denotes 0, 1, 2, 3 or 4;} each R⁴, if present, is independently selected from the group consisting of R⁵ and R⁶; each R⁵ is independently selected from the group consisting of -OR⁶, -NR⁶R⁶, halogen, -CN, -C(=O)R⁶, -C(=O)OR⁶, -C(=O)NR⁶R⁶, -NHC(=O)OR⁶ and the bivalent substituent =O; each R⁶ is independently selected from the group consisting of hydrogen, C_1-6alkyl, 10 C_1-6alkoxy, C_1-6haloalkyl, C_3-10cycloalkyl, C_4-10cycloalkenyl, 4-11 membered heterocyclyl, w_{herein the C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C3-10cycloalkyl, C4-10cycloalkenyl and 4-11 membered} heterocyclyl, are each independently optionally substituted with one or more, identical or different R⁷ and/or R⁸; each R⁷ is independently selected from the group consisting of -OH, -NH₂, -NHR⁸, -NR⁸R⁸,15 halogen, -CN, and C_1-6alkoxy; each R⁸ is independently selected from the group consisting of C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C3-10cycloalkyl, C4-10cycloalkenyl, and 4-11 membered heterocyclyl, wherein the C1- 6_{alkyl, C1-6alkoxy, C1-6haloalkyl, C3-10cycloalkyl, C4-10cycloalkenyl, and 4-11 membered heterocyclyl,} _{are each independently optionally substituted with one or more, identical or different R9;} 20 each R⁹ is -OH, halogen or C1-6alkoxy; or a salt thereof. I_{n another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof, wherein the ring} and is optionally substituted with one or two, identical or different R³. 9 12-0519-WO-1 I_{n another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof, wherein A1, A2,} _{A3, V and W form a triazole.} _{In another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof, wherein the ring} 5_{and is optionally substituted with one or two R3.} _{In another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof, wherein each R3,} if present, is C1-6alkyl. I_{n another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof, wherein each R3,} if present, is Me, Et or Pr. _{10 In another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof, wherein each R3,} if present, is Me. I_{n another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof, wherein q is 1 and} R³ is Me. I_{n another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof, wherein q is 0.} _{15 In another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof wherein p is 1.} _{In another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof wherein p is 2.} _{In another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof wherein p is 2 and} each R¹ is independently selected from the group consisting of Me, -CFH2, -CF2H, -CF3, -CFMeH, -CFMe2, -CF2Me, F. _{20 In another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof wherein p is 2 and} each R¹ is independently selected from the group consisting of Me, -CF2H, -CF3, F. I_{n another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof, wherein R2 is Me.} _{In another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof, wherein R2 is Me,} _{25 p is 2 and R1 is independently selected from the group consisting of Me, -CFH2, -CF2H, -CF3, -CFMeH,} -CFMe₂, -CF₂Me, and F. I_{n another aspect, the SOS1 inhibitor is acompound of formula (I), or a salt thereof, wherein ring system} _{B is selected from C3-10cycloalkyl, and C4-10cycloalkenyl.} _{In another aspect, the SOS1 inhibitor is acompound of formula (I), or a salt thereof, wherein ring system} _{30 B is selected from C5-7cycloalkyl, and C5-7cycloalkenyl.} 10 In another aspect, the SOS 1 inhibitor is a compound of formula (I), or a salt thereof, wherein ring system B is selected from cyclohexyl, and cyclohexenyl.

In another aspect, the SOS1 inhibitor is a compound of the formula (I), or a salt thereof, wherein ring system B is cyclohexyl. In another aspect, the SOS 1 inhibitor is a compound of formula (I), or a salt thereof, wherein ring system B is cyclohexenyl.

In another aspect, the SOS 1 inhibitor is a compound of formula (I), or a salt thereof, wherein ring system B is a 4-13 membered heterocyclyl.

In another aspect, the SOS 1 inhibitor is a compound of formula (I), or a salt thereof, wherein ring system B is piperidine.

In another aspect, the SOS 1 inhibitor is a compound of formula (I), or a salt thereof, wherein ring system B is selected from the group consisting of

wherein ring system B can be attached to the compound of formula (I) and to R⁴, if present, at any ring position by removal of a hydrogen atom.

In another aspect, the invention relates to the compound of the formula (I), or a salt thereof, wherein ring system B is selected from the group consisting of wherein ring system B can be attached to the compound of formula (I) and R⁴, if present, at any ring position by removal of a hydrogen atom.

In another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof, wherein r is 0, 1, 2, or 3.

In another aspect, the S0S1 inhibitor is a compound of formula (I), or a salt thereof, wherein r is 0, 1, or 2.

In another aspect, the S0S1 inhibitor is a compound of formula (I), or a salt thereof, wherein r is 0 or

In another aspect, the S0S1 inhibitor is a compound of formula (I), or a salt thereof, wherein r is 0. 12-0519-WO-1 I_{n another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof, wherein r is 1.} _{In another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof wherein each R5 is} independently selected from the group consisting of -OR⁶, -NR⁶R⁶, halogen, and -CN; each R⁶ is independently selected from the group consisting of hydrogen, C_1-6alkyl, 5 C_1-6alkoxy, C_1-6haloalkyl, C_3-10cycloalkyl, C_4-10cycloalkenyl, 4-11 membered heterocyclyl, w_{herein the C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C3-10cycloalkyl, C4-10cycloalkenyl and} _{4-11 membered heterocyclyl, are each independently optionally substituted with one or more, identical} or different R⁷ and/or R⁸. I_{n another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof wherein} 10 each R⁵ is independently selected from the group consisting of -OR⁶, -NR⁶R⁶, halogen, and -CN; each R⁶ is independently selected from the group consisting of hydrogen, C_1-6alkyl, C_3-10cycloalkyl, 4-11 membered heterocyclyl, wherein the C_1-6alkyl, C_3-10cycloalkyl and 4-11 m_{embered heterocyclyl, are each independently optionally substituted with one or more, identical or} different R⁷ and/or R⁸; 15 each R⁷ is independently selected from the group consisting of -OH, halogen, and C1-6alkoxy; each R⁸ is independently selected from the group consisting of C1-6alkyl, C3-10cycloalkyl, and 4-11 m_{embered heterocyclyl, wherein the C1-6alkyl, C3-10cycloalkyl, and 4-11 membered heterocyclyl, are} _{each independently optionally substituted with one or more, identical or different R9;} 20 each R⁹ is -OH. I_{n another aspect, the SOS1 inhibitor is a compound of formula (I), or a salt thereof, wherein each R4,} if present, is independently selected from the group consisting of , 13

In one embodiment, the SOS1 inhibitor relates to compounds of formula (I) wherein each R¹ is independently selected from the group consisting of Me, -CFH2, -CF2H, -CF3, -CFMeH, -CFMe₂, -CF₂Me, F; p is 2;

R² is Me;

V is nitrogen (-N=) or carbon

W is nitrogen (-N=) or carbon at least one of V and W is a nitrogen (-N=); 12-0519-WO-1 A_{1, A2 and A3 are each independently selected from nitrogen (-N=) or carbon (=CH-); at least one of} _{A1, A2 and A3 is a nitrogen (-N=)} and is a single or double bond; q_{is 0,} _{5 ring system B is a 4-13 membered heterocyclyl,} _{r denotes 0, 1, 2, or 3} each R⁴, if present, is independently selected from the group consisting of R⁵ and R⁶; each R⁵ is independently selected from the group consisting of -OR⁶, and -NR⁶R⁶; each R⁶ is independently selected from the group consisting of hydrogen, C_1-6alkyl, 10 C_3-10cycloalkyl, 4-11 membered heterocyclyl, wherein the C_1-6alkyl, C_3-10cycloalkyl and 4-11 m_{embered heterocyclyl, are each independently optionally substituted with one or more, identical or} different R⁷ and/or R⁸; each R⁷ is independently selected from the group consisting of -OH, halogen, and C1-6alkoxy; 15 each R⁸ is independently selected from the group consisting of C1-6alkyl, C3-10cycloalkyl, and 4-11 m_{embered heterocyclyl, wherein the C1-6alkyl, C3-10cycloalkyl, and 4-11 membered heterocyclyl, are} _{each independently optionally substituted with one or more, identical or different R9;} each R⁹ is -OH; or a salt thereof. _{20 In a further embodiment, the SOS1 inhibitor relates to compounds of formula (I) wherein} _{each R1 is independently selected from the group consisting of Me, -CFH2, -CF2H, -CF3,} -CFMeH, -CFMe2, -CF2Me, F; p_{is 2;} R² is Me; _{25 V is nitrogen (-N=) or carbon} W_{is nitrogen (-N=) or carbon} one of V and W is a nitrogen (-N=); A_{1, A2 and A3 are each independently selected from nitrogen (-N=) or carbon (=CH-); two of A1, A2} _{and A3 are a nitrogen (N)} 30 and is a single or double bond; q_{is 0,ring system B is a 4-13 membered heterocyclyl,} _{r denotes 0, 1, 2, or 3} each R⁴, if present, is independently selected from the group consisting of R⁵ and R⁶; 17 12-0519-WO-1 each R⁵ is independently selected from the group consisting of -OR⁶, and -NR⁶R⁶; each R⁶ is independently selected from the group consisting of hydrogen, C_1-6alkyl, C_3-10cycloalkyl, and 4-11 membered heterocyclyl, wherein the C_1-6alkyl, C_3-10cycloalkyl and 4-11 membered heterocyclyl, are each independently optionally substituted with one or more, identical or 5 different R⁷ and/or R⁸; each R⁷ is independently selected from the group consisting of -OH, halogen, and C_1-6alkoxy; each R⁸ is independently selected from the group consisting of C_1-6alkyl, C_3-10cycloalkyl, and 4-11 m_{embered heterocyclyl, wherein the C1-6alkyl, C3-10cycloalkyl, and 4-11 membered heterocyclyl, are}10 each independently optionally substituted with one or more, identical or different R⁹; each R⁹ is -OH; or a salt thereof. I_{n a further embodiment, the SOS1 inhibitor relates to compounds of formula (I) wherein} _{each R1 is independently selected from the group consisting of Me, -CFH2, -CF2H, -CF3,}15 -CFMeH, -CFMe₂, -CF₂Me, F; p_{is 2;} R² is Me; V_{is nitrogen (-N=);} _{W is carbon} _{20 A1 and A2 are independently nitrogen (-N=) or carbon (=CH-);} _{A3 is nitrogen (-N=);} _{q is 0,} _{ring system B is a 4-13 membered heterocyclyl,} _{r denotes 0, 1, 2, or 3} 25 each R⁴, if present, is independently selected from the group consisting of R⁵ and R⁶; each R⁵ is independently selected from the group consisting of -OR⁶, and -NR⁶R⁶; each R⁶ is independently selected from the group consisting of hydrogen, C_1-6alkyl, C_3-10cycloalkyl, and 4-11 membered heterocyclyl, wherein the C_1-6alkyl, C_3-10cycloalkyl and 4-11 membered heterocyclyl, are each independently optionally substituted with one or more, identical or30 different R⁷ and/or R⁸; each R⁷ is independently selected from the group consisting of -OH, halogen, and C1-6alkoxy; each R⁸ is independently selected from the group consisting of C1-6alkyl, C3-10cycloalkyl, and 4-11 m_{embered heterocyclyl, wherein the C1-6alkyl, C3-10cycloalkyl, and 4-11 membered heterocyclyl, are}35 each independently optionally substituted with one or more, identical or different R⁹; 18 12-0519-WO-1 each R⁹ is -OH; or a salt thereof. I_{n a further embodiment, the SOS1 inhibitor relates to compounds of formula (I) wherein} _{each R1 is independently selected from the group consisting of -CF2H, -CF3, -and F;} _{5 p is 2;} R² is Me; V_{is nitrogen (-N=);} _{W is carbon} A_{1 and A2 are independently nitrogen (-N=) or carbon (=CH-);} _{10 A3 is nitrogen (-N=);} _{q is 0,} _{ring system B is a 5-6 membered heterocyclyl,} _{r denotes 1 or 2} each R⁴, is independently selected from the group consisting of R⁵ and R⁶; 15 each R⁵ is independently selected from the group consisting of -OR⁶, and -NR⁶R⁶; each R⁶ is independently selected from the group consisting of hydrogen, C_1-6alkyl, C_3-10cycloalkyl, and 4-11 membered heterocyclyl, wherein the C_1-6alkyl, C_3-10cycloalkyl and 4-11 membered heterocyclyl, are each independently optionally substituted with one or more, identical or different R⁷ and/or R⁸; 20 each R⁷ is independently selected from the group consisting of -OH, halogen, and C_1-6alkoxy; each R⁸ is independently selected from the group consisting of C_1-6alkyl, C_3-10cycloalkyl, and 4-11 m_{embered heterocyclyl, wherein the C1-6alkyl, C3-10cycloalkyl, and 4-11 membered heterocyclyl, are} each independently optionally substituted with one or more, identical or different R⁹; 25 each R⁹ is -OH; or a salt thereof. I_{n a further embodiment, the SOS1 inhibitor relates to compounds of formula (I) wherein} _{each R1 is independently selected from the group consisting of -CF2H, -CF3, -and F;} _{p is 2;} 30 R² is Me; V_{is nitrogen (-N=);} _{W is carbon} A_{1 and A2 are independently nitrogen (-N=) or carbon (=CH-);} 19 12-0519-WO-1 A_{3 is nitrogen (-N=);} _{q is 0,} _{ring system B is a 5-6 membered heterocyclyl,} _{r is 1} 5 R⁴ is R⁶; R⁶ is a 4-11 membered heterocyclyl, wherein the 4-11 membered heterocyclyl, is optionally substituted with one or more, identical or different R⁷ and/or R⁸; each R⁷ is independently selected from the group consisting of -OH, halogen, and C1-6alkoxy; 10 each R⁸ is independently selected from the group consisting of C_1-6alkyl, C_3-10cycloalkyl, and 4-11 m_{embered heterocyclyl, wherein the C1-6alkyl, C3-10cycloalkyl, and 4-11 membered heterocyclyl, are} each independently optionally substituted with one or more, identical or different R⁹; each R⁹ is -OH; or a salt thereof. _{15 In a further embodiment, the SOS1 inhibitor relates to compounds of formula (I) wherein} _{each R1 is independently selected from the group consisting of -CF2H, -CF3, -and F;} _{p is 2;} R² is Me; V_{is nitrogen (-N=);} _{20 W is carbon} A_{1 and A2 are independently nitrogen (-N=) or carbon (=CH-);} _{A3 is nitrogen (-N=);} _{q is 0,} _{ring system B is a 5-6 membered heterocyclyl,} _{25 r is 1} R⁴ is R⁶; R⁶ is a 4-11 membered heterocyclyl, wherein the 4-11 membered heterocyclyl, is optionally substituted with one or more, identical or different R⁷ and/or R⁸; each R⁷ is -OH; 30 each R⁸ is independently selected from the group consisting of C_1-6alkyl, and C_3-10cycloalkyl; or a salt thereof. In a further embodiment, the invention relates to compounds of formula (I) wherein e_{ach R1 is independently selected from the group consisting of -CF2H, -CF3, -and F;} _{p is 2;} 35 R² is Me; 20 12-0519-WO-1 V_{is nitrogen (-N=);} _{W is carbon} A_{1 and A2 are independently nitrogen (-N=) or carbon (=CH-);} _{A3 is nitrogen (-N=);} _{5 q is 0,} _{ring system B is a C3-10cycloalkyl,} _{r is 1} R⁴ is R⁶; R⁶ is a 4-11 membered heterocyclyl, wherein the 4-11 membered heterocyclyl, is optionally substituted10 with one or more, identical or different R⁷ and/or R⁸; each R⁷ is independently selected from the group consisting of -OH, halogen, and C1-6alkoxy; each R⁸ is independently selected from the group consisting of C_1-6alkyl, C_3-10cycloalkyl, and 4-11 m_{embered heterocyclyl, wherein the C1-6alkyl, C3-10cycloalkyl, and 4-11 membered heterocyclyl, are}15 each independently optionally substituted with one or more, identical or different R⁹; each R⁹ is -OH; or a salt thereof. In a further embodiment, the invention relates to compounds of formula (I) wherein e_{ach R1 is independently selected from the group consisting of -CF2H, -CF3, -and F;} _{20 p is 2;} R² is Me; V_{is nitrogen (-N=);} _{W is carbon} A_{1 and A2 are independently nitrogen (-N=) or carbon (=CH-);} _{25 A3 is nitrogen (-N=);} _{q is 0,} _{ring system B is a C3-10cycloalkyl,} _{r is 1} R⁴ is R⁶; 30 R⁶ is a 4-11 membered heterocyclyl, wherein the 4-11 membered heterocyclyl, is optionally substituted with one or more, identical or different R⁷ and/or R⁸; each R⁷ is -OH; each R⁸ is independently selected from the group consisting of C_1-6alkyl, and C₃₁₀cycloalkyl; 21 or a salt thereof.

In a preferred embodiment the compound of formula (I) is selected from pharmaceutically acceptable salt thereof.

It is to be understood that any two or more aspects and/or preferred embodiments of formula (I) — or subformulas thereof — may be combined in any way leading to a chemically stable structure to obtain further aspects of formula (I) — or subformulas thereof.

Preferred embodiments of the SOS1 inhibitor are example compounds El to E86.

Preferred embodiments of the SOS1 inhibitor are example compounds El to E86 and/or the pharmaceutically acceptable salts thereof.

The term “SOS1 inhibitor” includes hydrates, solvates, polymorphs, metabolites, derivatives, stereoisomers and prodrugs of a compound of formula (I) (including all embodiments thereof).

In an aspect, the SOS 1 inhibitor is a hydrate of a compound of formula (I) (including all aspects thereof). In another aspect, the SOS1 inhibitor is a solvate of a compound of formula (I) (including all embodiments thereof).

Compounds of formula (I) (including all embodiments thereof) which e.g. bear ester groups are potential prodrugs the ester being cleaved under physiological conditions and are also understood as SOS1 inhibitors.

In another aspect, the SOS1 inhibitor is a pharmaceutically acceptable salt of a compound of formula (I) (including all embodiments thereof).

In another aspect, the SOS1 inhibitor is a pharmaceutically acceptable salt of a compound of formula (I) (including all embodiments thereof) with anorganic or organic acids or bases.

Accordingly, in another aspect the SOS 1 inhibitor further relates to compounds of formula (I) as defined herein or pharmaceutically acceptable salts thereof for use as a medicament. Other aspects of the SO SI inhibitor will become apparent to the person skilled in the art directly from the foregoing and following description and examples.

The combination of all aspect in respect of the nature of zongertinib or the pharmaceutically acceptable salt thereof as described herein with all aspects in respect of the nature of the SOS1 inhibitor as described herein results in specific combinations which shall all be deemed to be specifically disclosed and to be embodiments of the invention and of all combinations, compositions, kits, methods, uses and compounds for use described herein. Preferred embodiments are combinations of aspects of zongertinib or the pharmaceutically acceptable salt thereof with aspects of SOS1 inhibitor. Definitions

Terms not specifically defined herein should be given the meanings that would be given to them by one of skill in the art in light of the disclosure and the context. As used in the specification, however, unless specified to the contrary, the following terms have the meaning indicated and the following conventions are adhered to:

The use of the prefix C_x.y, wherein x and y each represent a positive integer (x < y), indicates that the chain or ring structure or combination of chain and ring structure as a whole, specified and mentioned in direct association, may consist of a maximum of y and a minimum of x carbon atoms.

The indication of the number of members in groups that contain one or more heteroatom(s) (e.g. heteroaryl, heteroarylalkyl, heterocyclyl, heterocycylalkyl) relates to the total number of atoms of all the ring members or the total of all the ring and carbon chain members. Obviously, a ring structure has at least three members.

In general, for groups comprising two or more subgroups (e.g. heteroarylalkyl, heterocycylalkyl, cycloalkylalkyl, arylalkyl) the last named subgroup is the radical attachment point, for example, the substituent aryl-Ci-ealkyl means an aryl group which is bound to a Ci-ealkyl group, the latter of which is bound to the core or to the group to which the substituent is attached.

In groups like HO, H2N, (O)S, (0)28, NC (cyano), HOOC, F3C or the like, the skilled artisan can see the radical attachment point(s) to the molecule from the free valences of the group itself.

Alkyl denotes monovalent, saturated hydrocarbon chains, which may be present in both straight-chain (unbranched) and branched form. If an alkyl is substituted, the substitution may take place independently of one another, by mono- or polysubstitution in each case, on all the hydrogen-carrying carbon atoms.

The term ”Ci 5alkyl“ includes for example H3C-, H3C-CH2-, H3C-CH2-CH2-, H3C-CH(CH3)-, H3C-CH2- CH2-CH2-, H₃C-CH₂-CH(CH₃)-, H₃C-CH(CH₃)-CH₂-, H₃C-C(CH₃)2-, H3C-CH2-CH2-CH2-CH2-, H₃C- CH₂-CH₂-CH(CH₃)-, H₃C-CH2-CH(CH3)-CH₂-, H₃C-CH(CH3)-CH2-CH₂-, H₃C-CH₂-C(CH3)2-, H₃C-C(CH₃)2-CH₂-, H₃C-CH(CH3)-CH(CH₃)- and H₃C-CH2-CH(CH₂CH3)-.

Further examples of alkyl are methyl (Me; -CH3), ethyl (Et; -CH2CH3), 1 -propyl (w-propyk n- Pr; -CH2CH2CH3), 2-propyl (z-Pr; iso -propyl; -CH(CH3)2), 1 -butyl (w-butyk zz-Bu; -CH2CH2CH2CH3), 2 -methyl- 1 -propyl (zso-butyl; z-Bu; -CH2CH(CH3)2), 2-butyl (sec-butyk sec-Bu -CH(CH3)CH2CH3), 2 -methyl -2 -propyl (tert-butyl; t-Bu; -C(CH3)3), 1 -pentyl (zz-pentyl; -CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3 -pentyl (-CH(CH2CH3)2), 3 -methyl- 1 -butyl (iso- pentyl; -CH2CH2CH(CH3)2), 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3 -methyl -2 -butyl

(-CH(CH3)CH(CH3)2), 2,2-dimethyl-l -propyl (zzeo-pentyl; -CTFC CTE^), 2 -methyl- 1 -butyl (-CH₂CH(CH3)CH₂CH3), 1 -hexyl (zz-hexyl; -CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH₂CH₂CH₂CH3), 3 -hexyl (-CH(CH₂CH3)(CH₂CH₂CH3)), 2-methyl-2-pentyl (-C(CH3)2CH₂CH₂CH3), 3 -methyl -2 -pentyl (-CH(CH3)CH(CH₃)CH₂CH₃), 4-methyl-2 -pentyl

(-CH(CH₃)CH₂CH(CH₃)₂), 3 -methyl-3 -pentyl (-C(CH₃)(CH₂CH₃)₂), 2-methyl-3 -pentyl

(-CH(CH₂CH₃)CH(CH₃)₂), 2.3-dimethyl-2-butyl (-C(CH₃)₂CH(CH₃)₂), 3 ,3 -dimethyl -2 -butyl

(-CH(CH₃)C(CH₃)3), 2,3 -dimethyl- 1 -butyl (-CH₂CH(CH3)CH(CH₃)CH₃), 2,2-dimethyl- 1 -butyl

(-CH₂C(CH₃)₂CH₂CH₃), 3.3-dimethyl-l-butyl (-CH₂CH₂C(CH3)3), 2-methyl- 1 -pentyl

(-CH₂CH(CH₃)CH₂CH₂CH₃), 3 -methyl- 1 -pentyl (-CH₂CH₂CH(CH₃)CH₂CH₃), 1-heptyl (w-heptyl), 2- methyl-1 -hexyl, 3 -methyl- 1 -hexyl, 2,2-dimethyl- 1 -pentyl, 2,3 -dimethyl- 1 -pentyl, 2,4-dimethyl-l- pentyl, 3,3-dimethyl-l-pentyl, 2,2,3-trimethyl-l-butyl, 3 -ethyl- 1 -pentyl, 1-octyl (w-octyl). 1-nonyl (w-nonyl); 1 -decyl (w-dccyl) etc.

By the terms propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl etc. without any further definition are meant saturated hydrocarbon groups with the corresponding number of carbon atoms, wherein all isomeric forms are included.

The above definition for alkyl also applies if alkyl is a part of another (combined) group such as for example C_x-yalkylamino or C_x.yalkyloxy.

Unlike alkyl, alkenyl consists of at least two carbon atoms, wherein at least two adjacent carbon atoms are joined together by a C-C double bond and a carbon atom can only be part of one C-C double bond. If in an alkyl as hereinbefore defined having at least two carbon atoms, two hydrogen atoms on adjacent carbon atoms are formally removed and the free valencies are saturated to form a second bond, the corresponding alkenyl is formed.

Examples of alkenyl are vinyl (ethenyl), prop-l-enyl, allyl (prop-2 -enyl), isopropenyl, but-l-enyl, but-

2-enyl, but-3-enyl, 2-methyl -prop-2 -enyl, 2-methyl-prop-l-enyl, 1 -methyl -prop-2 -enyl, 1 -methyl -prop- l-enyl, 1 -methylidenepropyl, pent- 1 -enyl, pent-2 -enyl, pent-3 -enyl, pent-4-enyl, 3 -methyl -but-3 -enyl,

3 -methyl -but-2 -enyl, 3-methyl-but-l-enyl, hex- 1 -enyl, hex-2 -enyl, hex-3 -enyl, hex-4-enyl, hex-5 -enyl, 2,3 -dimethyl -but-3 -enyl, 2,3 -dimethyl -but-2 -enyl, 2-methylidene-3 -methylbutyl, 2,3 -dimethyl -but- 1 - enyl, hexa-1, 3-dienyl, hexa- 1,4-dienyl, penta- 1,4-dienyl, penta-1, 3-dienyl, buta-1, 3-dienyl, 2,3- dimethylbuta- 1,3 -diene etc.

By the generic terms propenyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl, heptadienyl, octadienyl, nonadienyl, decadienyl etc. without any further definition are meant all the conceivable isomeric forms with the corresponding number of carbon atoms, i.e. propenyl includes prop-l-enyl and prop-2 -enyl, butenyl includes but-l-enyl, but-2 -enyl, but-3 -enyl, 1 -methyl -prop-l- enyl, 1 -methyl -prop-2 -enyl etc.

Alkenyl may optionally be present in the cis or trans or E or Z orientation with regard to the double bond(s).

The above definition for alkenyl also applies when alkenyl is part of another (combined) group such as for example in C_x.yalkenylamino or C_x.yalkenyloxy.

By heteroatoms are meant oxygen, nitrogen and sulphur atoms. Haloalkyl is derived from the previously defined alkyl by replacing one or more hydrogen atoms of the hydrocarbon chain independently of one another by halogen atoms, which may be identical or different. If a haloalkyl is to be further substituted, the substitutions may take place independently of one another, in the form of mono- or poly substitutions in each case, on all the hydrogen-carrying carbon atoms.

Examples of haloalkyl are -CF₃, -CHF₂, -CH₂F, -CF₂CF₃, -CHFCF₃, -CH₂CF₃, -CF₂CH₃, -CHFCH₃, -CF₂CF₂CF₃, -CF₂CH₂CH₃, -CHFCH₂CH₃, -CHFCH₂CF₃ etc.

Halogen relates to fluorine, chlorine, bromine and/or iodine atoms.

Cycloalkyl is made up of the subgroups monocyclic hydrocarbon rings, bicyclic hydrocarbon rings and spiro-hydrocarbon rings. The systems are saturated. In bicyclic hydrocarbon rings two rings are joined together so that they have at least two carbon atoms in common. In spiro-hydrocarbon rings one carbon atom (spiroatom) belongs to two rings together.

If a cycloalkyl is to be substituted, the substitutions may take place independently of one another, in the form of mono- or polysubstitutions in each case, on all the hydrogen-carrying carbon atoms. Cycloalkyl itself may be linked as a substituent to the molecule via every suitable position of the ring system.

Examples of cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclo[2.2.0]hexyl, bicyclo[3.2.0]heptyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octyl, bicyclo[4.3.0]nonyl (octahydroindenyl), bicyclo[4.4.0]decyl (decahydronaphthyl), bicyclo[2.2.1]heptyl (norbomyl), bicyclo[4.1.0]heptyl (norcaranyl), bicyclo [3. 1. Ijheptyl (pinanyl), spiro[2.5]octyl, spiro[3.3]heptyl etc. If the free valency of a cycloalkyl is saturated, then an alicycle is obtained.

Cycloalkenyl is also made up of the subgroups monocyclic hydrocarbon rings, bicyclic hydrocarbon rings and spiro-hydrocarbon rings. However, the systems are unsaturated, i.e. there is at least one C- C double bond but no aromatic system. If in a cycloalkyl as hereinbefore defined two hydrogen atoms at adjacent cyclic carbon atoms are formally removed and the free valencies are saturated to form a second bond, the corresponding cycloalkenyl is obtained.

If a cycloalkenyl is to be substituted, the substitutions may take place independently of one another, in the form of mono- or polysubstitutions in each case, on all the hydrogen-carrying carbon atoms. Cycloalkenyl itself may be linked as a substituent to the molecule via every suitable position of the ring system.

Examples of cycloalkenyl are cycloprop- 1-enyl, cycloprop-2 -enyl, cyclobut-l-enyl, cyclobut-2-enyl, cyclopent- 1-enyl, cyclopent-2 -enyl, cyclopent-3 -enyl, cyclohex- 1 -enyl, cyclohex-2 -enyl, cyclohex-3 - enyl, cyclohept- 1 -enyl, cyclohept-2-enyl, cyclohept-3-enyl, cyclohept-4-enyl, cyclobuta-1, 3-dienyl, cyclopenta- 1,4-dienyl, cyclopenta-1, 3-dienyl, cyclopenta-2, 4-dienyl, cyclohexa- 1,3 -dienyl, cyclohexa- 1, 5-dienyl, cyclohexa-2, 4-dienyl, cyclohexa- 1,4-dienyl, cyclohexa-2, 5-dienyl, bicyclo[2.2.1]hepta-2,5- dienyl (norboma-2, 5-dienyl), bicyclo[2.2.1]hept-2-enyl (norbomenyl), spiro[4,5]dec-2-enyl etc. If the free valency of a cycloalkenyl is saturated, then an unsaturated alicycle is obtained.

Heterocyclyl denotes ring systems, which are derived from the previously defined cycloalkyl and cycloalkenyl by replacing one or more of the groups -CH2- independently of one another in the hydrocarbon rings by the groups -O-, -S- or -NH- or by replacing one or more of the groups =CH- by the group =N-, wherein a total of not more than five heteroatoms may be present, at least one carbon atom must be present between two oxygen atoms and between two sulphur atoms or between an oxygen and a sulphur atom and the ring as a whole must have chemical stability. Heteroatoms may optionally be present in all the possible oxidation stages (sulphur a sulphoxide -SO-, sulphone -SO2-; nitrogen a N-oxide). In a heterocyclyl there is no heteroaromatic ring, i.e. no heteroatom is part of an aromatic system.

A direct result of the derivation from cycloalkyl and cycloalkenyl is that heterocyclyl is made up of the subgroups monocyclic heterorings, bicyclic heterorings, tricyclic heterorings and spiro- heter orings, which may be present in saturated or unsaturated form.

By unsaturated is meant that there is at least one double bond in the ring system in question, but no heteroaromatic system is formed. In bicyclic heterorings two rings are linked together so that they have at least two (hetero)atoms in common. In spiro-heterorings one carbon atom (spiroatom) belongs to two rings together.

If a heterocyclyl is substituted, the substitutions may take place independently of one another, in the form of mono- or polysubstitutions in each case, on all the hydrogen-carrying carbon and/or nitrogen atoms. Heterocyclyl itself may be linked as a substituent to the molecule via every suitable position of the ring system. Substituents on heterocyclyl do not count for the number of members of a heterocyclyl.

Examples of heterocyclyl are tetrahydrofuryl, pyrrolidinyl, pyrrolinyl, imidazolidinyl, thiazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperidinyl, piperazinyl, oxiranyl, aziridinyl, azetidinyl, 1,4- dioxanyl, azepanyl, diazepanyl, morpholinyl, thiomorpholinyl, homomorpholinyl, homopiperidinyl, homopiperazinyl, homothiomorpholinyl, thiomorpholinyl-S'-oxidc. thiomorpholinyl -S' .S'-dioxidc. 1,3- dioxolanyl, tetrahydropyranyl, tetrahydrothiopyranyl, [l,4]-oxazepanyl, tetrahydrothienyl, homothiomorpholinyl -S' .S'-dioxidc. oxazolidinonyl, dihydropyrazolyl, dihydropyrrolyl, dihydropyrazinyl, dihydropyridyl, dihydro-pyrimidinyl, dihydrofuryl, dihydropyranyl, tctrahydrothicnyl-S'-oxidc. tctrahydrothicnyl-S'..S'-dioxidc. homothiomorpholinyl-S'-oxidc. 2,3- dihydroazet, 2//-pyrrolyl. 4//-pyranyl. 1,4-dihydropyridinyl, 8-aza-bicyclo[3.2.1]octyl, 8-aza- bicyclo [5.1.0]octyl, 2-oxa-5 -azabicyclo [2.2. 1 ]heptyl, 8-oxa-3 -aza-bicyclo [3.2.1] octyl,

3.8-diaza-bicyclo [3.2.1] octyl, 2,5 -diaza-bicyclo [2.2. 1 ]heptyl, 1 -aza-bicyclo [2.2 ,2]octyl,

3.8-diaza-bicyclo[3.2.1]octyl, 3,9-diaza-bicyclo[4.2.1]nonyl, 2,6-diaza-bicyclo[3.2.2]nonyl, 1,4-dioxa- spiro[4.5]decyl, l-oxa-3,8-diaza-spiro[4.5]decyl, 2,6-diaza-spiro[3.3]heptyl, 2,7-diaza-spiro[4.4]nonyl, 2,6-diaza-spiro[3.4]octyl, 3,9-diaza-spiro[5.5]undecyl, 2.8-diaza-spiro[4,5]decyl etc. Further examples are the structures illustrated below, which may be attached via each hydrogencarrying atom (exchanged for hydrogen):

If the free valency of a heterocyclyl is saturated, then a heterocycle is obtained.

Heteroaryl denotes monocyclic heteroaromatic rings or polycyclic rings with at least one heteroaromatic ring, which compared with the corresponding aryl or cycloalkyl (cycloalkenyl) contain, 12-0519-WO-1 instead of one or more carbon atoms, one or more identical or different heteroatoms, selected independently of one another from among nitrogen, sulphur and oxygen, wherein the resulting group m_{ust be chemically stable. The prerequisite for the presence of heteroaryl is a heteroatom and a} heteroaromatic system. 5_{If a heteroaryl is to be substituted, the substitutions may take place independently of one another, in} _{the form of mono- or polysubstitutions in each case, on all the hydrogen-carrying carbon and/or nitrogen} _{atoms. Heteroaryl itself may be linked as a substituent to the molecule via every suitable position of} _{the ring system, both carbon and nitrogen. Substituents on heteroaryl do not count for the number of} members of a heteroaryl. _{10 Examples of heteroaryl are furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl,} pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxadiazolyl, thiadiazolyl, pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, triazinyl, pyridyl-N-oxide, pyrrolyl-N-oxide, pyrimidinyl-N-oxide, pyridazinyl-N-oxide, pyrazinyl-N-oxide, imidazolyl-N-oxide, isoxazolyl-N-oxide, oxazolyl-N-oxide, thiazolyl-N-oxide, oxadiazolyl-N-oxide, thiadiazolyl-N-oxide, triazolyl-N-oxide, tetrazolyl-N-oxide, indolyl, isoindolyl, 15 benzofuryl, benzothienyl, benzoxazolyl, benzothiazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolyl, indazolyl, isoquinolinyl, quinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, benzotriazinyl, indolizinyl, oxazolopyridyl, imidazopyridyl, naphthyridinyl, benzoxazolyl, pyridopyridyl, pyrimidopyridyl, purinyl, pteridinyl, benzothiazolyl, imidazopyridyl, imidazothiazolyl, quinolinyl-N-oxide, indolyl-N-oxide, isoquinolyl-N-oxide, quinazolinyl-N-oxide,20 quinoxalinyl-N-oxide, phthalazinyl-N-oxide, indolizinyl-N-oxide, indazolyl-N-oxide, benzothiazolyl- N-oxide, benzimidazolyl-N-oxide etc. F_{urther examples are the structures illustrated below, which may be attached via each hydrogen-} carrying atom (exchanged for hydrogen): 38

Preferably, heteroaryls are 5-6 membered monocyclic or 9-10 membered bicyclic, each with 1 to 4 heteroatoms independently selected from oxygen, nitrogen and sulfur.

If the free valency of a heteroaryl is saturated, a heteroarene is obtained. single or double bond.

By substituted is meant that a hydrogen atom which is bound directly to the atom under consideration, is replaced by another atom or another group of atoms (substituent). Depending on the starting conditions (number of hydrogen atoms) mono- or polysubstitution may take place on one atom. Substitution with a particular substituent is only possible if the permitted valencies of the substituent and of the atom that is to be substituted correspond to one another and the substitution leads to a stable compound (i.e. to a compound which is not converted spontaneously, e.g. by rearrangement, cyclisation or elimination).

Bivalent substituents such as =S, =NR, =NOR, =NNRR, =NN(R)C(O)NRR, =N2 or the like, may only be substituents on carbon atoms, whereas the bivalent substituents =0 and =NR may also be a substituent on sulphur. Generally, substitution may be carried out by a bivalent substituent only at ring systems and requires replacement of two geminal hydrogen atoms, i.e. hydrogen atoms that are bound to the same carbon atom that is saturated prior to the substitution. Substitution by a bivalent substituent is therefore only possible at the group -CH2- or sulphur atoms (=0 group or =NR group only, one or two =0 groups possible or, e.g., group and group, each group replacing a free electron pair) of a ring system.

Stereochemistry/solvates/hydrates: Unless specifically indicated, throughout the specification and appended claims, a given chemical formula or name shall encompass tautomers and all stereo, optical and geometrical isomers (e.g. enantiomers, diastereomers, cis/trans/Z isomers, etc.) and racemates thereof as well as mixtures in different proportions of the separate enantiomers, mixtures of diastereomers, or mixtures of any of the foregoing forms where such isomers and enantiomers exist, and solvates thereof such as for instance hydrates.

Unless specifically indicated, also “pharmaceutically acceptable salts” as defined in more detail below shall encompass solvates thereof such as for instance hydrates.

In general, substantially pure stereoisomers can be obtained according to synthetic principles known to a person skilled in the field, e.g. by separation of corresponding mixtures, by using stereochemically pure starting materials and/or by stereoselective synthesis. It is known in the art how to prepare optically active forms, such as by resolution of racemic forms or by synthesis, e.g. starting from optically active starting materials and/or by using chiral reagents.

Enantiomerically pure compounds of this invention or intermediates may be prepared via asymmetric synthesis, for example by preparation and subsequent separation of appropriate diastereomeric compounds or intermediates which can be separated by known methods (e.g. by chromatographic separation or crystallization) and/or by using chiral reagents, such as chiral starting materials, chiral catalysts or chiral auxiliaries.

Further, it is known to the person skilled in the art how to prepare enantiomerically pure compounds from the corresponding racemic mixtures, such as by chromatographic separation of the corresponding racemic mixtures on chiral stationary phases, or by resolution of a racemic mixture using an appropriate resolving agent, e.g. by means of diastereomeric salt formation of the racemic compound with optically active acids or bases, subsequent resolution of the salts and release of the desired compound from the salt, or by derivatization of the corresponding racemic compounds with optically active chiral auxiliary reagents, subsequent diastereomer separation and removal of the chiral auxiliary group, or by kinetic resolution of a racemate (e.g. by enzymatic resolution); by enantioselective crystallization from a conglomerate of enantiomorphous crystals under suitable conditions, or by (fractional) crystallization from a suitable solvent in the presence of an optically active chiral auxiliary.

Salts: The phrase “pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgement, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, and commensurate with a reasonable benefit/risk ratio.

As used herein “pharmaceutically acceptable salts” refers to derivatives of the disclosed compounds wherein the parent compound is modified by making acid or base salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. For example, such salts include salts from benzenesulfonic acid, benzoic acid, citric acid, ethanesulfonic acid, fumaric acid, gentisic acid, hydrobromic acid, hydrochloric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, 4-methyl-benzenesulfonic acid, phosphoric acid, salicylic acid, succinic acid, sulfuric acid and tartaric acid.

Further pharmaceutically acceptable salts can be formed with cations from ammonia, L-arginine, calcium, 2,2’-iminobisethanol, L-lysine, magnesium, A-methyl-D-glucamine, potassium, sodium and tris(hydroxymethyl)-aminomethane .

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base form of these compounds with a sufficient amount of the appropriate base or acid in water or in an organic diluent like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile, or a mixture thereof.

Salts of other acids than those mentioned above which for example are useful for purifying or isolating the compounds of the present invention (e.g. trifluoro acetate salts), also comprise a part of the invention.

By a therapeutically effective amount for the purposes of this invention is meant a quantity of substance that is capable of obviating symptoms of illness or of preventing or alleviating these symptoms, or which prolong the survival of a treated patient.

RAS-family proteins are meant to include KRAS (V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog), NRAS (neuroblastoma RAS viral oncogene homolog), HRAS (Harvey murine sarcoma virus oncogene) and MRAS (muscle RAS oncogene homolog) and any mutants thereof.

A SOS1 inhibitor compound is a compound, which binds to SOS1 and thereby prevents the SOS1 mediated nucleotide exchange and subsequently reduces the levels of RAS in its GTP bound form. More specifically, a SO SI inhibitor compound shows a pharmacological inhibition of the binding of the catalytic site of SOS1 to RAS-family proteins. Thus, such a compound interacts with SOS1, e.g. the catalytic site on SOS1, and reduces the level of binding to the RAS-family protein in relation to said binding without addition of a SOS1 inhibitor compound. Accordingly, it is envisaged that a SOS1 inhibitor compound at least reduces the level of binding to the RAS-family protein about 5 %, 10 %, 20 %, 30 %, 40 %, 50 %, 60 %, 70 %, 80 %, 90 % or even 100 % when compared to the binding that is achieved without the addition of said inhibitor compound. Suitable test systems to measure the binding to the catalytic site of SOS1 are disclosed herein. Said compound may be chemically synthesized (e.g. a small molecule) or microbiologically produced (e.g. a monoclonal antibody) and/or comprised in, for example, samples, e.g., cell extracts from, e.g., plants, animals or microorganisms. Preferably, the S0S1 inhibitor compound is a small molecule.

Combination Therapy

It is to be understood that the combinations, compositions, kits, methods, uses or compounds for use according to this invention may envisage the simultaneous, concurrent, sequential, successive, alternate or separate administration of the active agents or components. It will be appreciated that zongertinib or the pharmaceutically acceptable salt thereof and the S0S1 inhibitor can be formulated either together or independently. For example, zongertinib or the pharmaceutically acceptable salt thereof and the SOS 1 inhibitor may be administered either as part of the same pharmaceutical composition/dosage form or, preferably, in separate pharmaceutical compositions/dosage forms.

Zongertinib or the pharmaceutically acceptable salt thereof and the S0S1 inhibitor thus may be formulated, alone or together, in suitable dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, excipients and/or vehicles appropriate for each route of administration. Typical pharmaceutical compositions for administering zongertinib or the pharmaceutically acceptable salt thereof and the S0S1 inhibitor, separately or jointly, include for example tablets, capsules, suppositories, solutions, e.g. solutions for injection and infusion, elixirs, emulsions or dispersible powders. Dosage forms and formulations of active ingredients are known in the art and further described herein.

In this context, “combination” or “combined” and grammatical variants thereof within the meaning of this invention include, but are not limited to, a product, product for use, use or method that results from the mixing or combining of more than one active agent, in particular zongertinib and the SOS 1 inhibitor as defined herein. The expressions “combination”, “combined” and grammatical variants thereof comprise both fixed (e.g. pharmaceutical composition) and non-fixed (e.g. free) combinations (e.g. kits), products, products for use, uses and methods, such as e.g. the simultaneous, concurrent, sequential, successive, alternate or separate use of zongertinib and the SOS 1 inhibitor as defined herein. As used herein, “combination” or “combined” and grammatical variants thereof refer in particular to a combination that takes place within the same line of treatment. The term “fixed combination” means that zongertinib and the SOS1 inhibitor as defined herein are both administered to a patient simultaneously in the form of a single entity or dosage form. The term “nonfixed combination” means that the active agents are both administered to a patient as separate entities or dosage forms either simultaneously, concurrently, sequentially, successively, alternatively or otherwise separately with no specific time limits.

The term “simultaneous” refers to the administration of both compounds/compositions at substantially the same time. This form of administration may also be referred to as “concomitant” administration. The term “concurrent” refers to administration of the active ingredients within the same general time period, for example on the same day(s) but not necessarily at the same time. The term “sequential” administration includes administration of one active ingredient during a first time period, for example over the course of a few hours, days or a week, using one or more doses, followed by administration of the other active ingredient during a second time period, for example over the course of a few hours, days or a week, using one or more doses. An overlapping schedule may also be employed, which includes administration of the active ingredients on different days over the treatment period, not necessarily according to a regular sequence. The term “successive” administration, alternatively, refers to an administration where the second administration step is carried out immediately once the administration of the first compounds has been finished. Alternate administration includes administration of one active ingredient during a time period, for example over the course of a few hours, days or a week, followed by administration of the other active ingredient during a subsequent period of time, for example over the course of a few hours, days or a week, and then repeating the pattern for one or more cycles, wherein the overall number of repeats depends on the chosen dosage regimen. Variations of these general administration forms may also be employed.

Accordingly, in another aspect, zongertinib or the pharmaceutically acceptable salt thereof and the SOS1 inhibitor are administered simultaneously, concurrently, sequentially, successively, alternately or separately.

In another aspect, zongertinib or the pharmaceutically acceptable salt thereof and SOS1 inhibitor are administered simultaneously.

In another aspect, zongertinib or the pharmaceutically acceptable salt thereof and the SOS1 inhibitor are administered concurrently.

In another aspect, zongertinib or the pharmaceutically acceptable salt thereof and the SOS1 inhibitor are administered sequentially.

In another aspect, zongertinib or the pharmaceutically acceptable salt thereof and the SOS1 inhibitor are administered successively.

In another aspect, zongertinib or the pharmaceutically acceptable salt thereof and the SOS1 inhibitor are administered alternately. In another aspect, zongertinib or the pharmaceutically acceptable salt thereof and the SOS1 inhibitor are administered separately.

In another aspect, zongertinib or the pharmaceutically acceptable salt thereof is administered before the SOS1 inhibitor.

In another aspect, the SOS1 inhibitor is administered immediately after zongertinib or the pharmaceutically acceptable salt thereof. In this context, “immediately after” means, e.g. 30 minutes, 1 hour, 2 hours, 3 hours or 4 hours after.

In another aspect, zongertinib or the pharmaceutically acceptable salt thereof is administered after the SOS1 inhibitor. Preferably, the treatment comprises one day when both zongertinib or the pharmaceutically acceptable salt thereof and the SOS 1 inhibitor are administered and zongertinib or the pharmaceutically acceptable salt thereof is administered after the SOS1 inhibitor.

Additional intervention

The combination of zongertinib, or a pharmaceutically acceptable salt thereof, and a SOS 1 inhibitor as described herein may be administered in combination with additional intervention selected from the group consisting of radiation, surgery, an additional therapeutic agent such as chemotherapy and a combination thereof.

In one aspect, the present invention relates to a use of the combination of zongertinib, or a pharmaceutically acceptable salt thereof, and a SOS 1 inhibitor as described herein in combination with a cytostatic and/or cytotoxic active substance and/or in combination with radiotherapy and/or in combination with surgery and/or in combination with chemotherapy and/or in combination with immunotherapy in the treatment and/or prevention of cancer.

For the treatment of diseases of oncological nature, anticancer agents may be combined with radiotherapy, e.g. irradiation treatment, and/or surgery. The combination of zongertinib, or a pharmaceutically acceptable salt thereof, and a SO SI inhibitor, as well as the relative compounds for use, methods of treatment, uses, pharmaceutical compositions and kits as described herein, can be used in combination with radiotherapy. For example, a cancer patient may receive radiotherapy before and/or after or simultaneously with receiving therapy with the combination of zongertinib, or a pharmaceutically acceptable salt thereof, and a SO SI inhibitor, as well as with the relative compounds for use, methods of treatment, uses, pharmaceutical compositions and kits as described herein.

In embodiments, the combination of zongertinib, or a pharmaceutically acceptable salt thereof, and a SOS1 inhibitor, as well as the relative compounds for use, methods of treatment, uses, pharmaceutical compositions and kits as described herein, are used as adjuvant therapy in combination with a surgical procedure. The combination of zongertinib, or a pharmaceutically acceptable salt thereof, and a SOS1 inhibitor, the relative compounds for use, methods of treatment, uses, pharmaceutical compositions and kits described herein may be administered for the purpose of diminishing the size of a tumor before surgical procedure (referred to as pre-operative adjuvant chemotherapy or neoadjuvant therapy), or may be administered after a surgical procedure for the purpose.

In embodiments, the combination of zongertinib, or a pharmaceutically acceptable salt thereof, and a SOS1 inhibitor, the relative compounds for use, methods of treatment, uses, pharmaceutical compositions and kits as described herein are administered in combination with a cytostatic and/or cytotoxic active substance and/or in combination with immunotherapy.

In embodiments, the combination of zongertinib, or a pharmaceutically acceptable salt thereof, and a SOS1 inhibitor, as well as the relative compounds for use, methods of treatment, uses, pharmaceutical compositions and kits as described herein may be used in combination with one or several other pharmacologically active substances such as state-of-the-art or standard-of-care compounds, such as e.g. cell proliferation inhibitors, anti -angiogenic substances, steroids or immune modulators/checkpoint inhibitors, and the like.

Pharmacologically active substances which may be administered in combination with the combination of zongertinib, or a pharmaceutically acceptable salt thereof, and a SO SI inhibitor, as well as the relative compounds for use, methods of treatment, uses, pharmaceutical compositions and kits as described herein, include, without being restricted thereto, hormones, hormone analogues and antihormones (e.g. tamoxifen, toremifene, raloxifene, fulvestrant, megestrol acetate, flutamide, nilutamide, bicalutamide, aminoglutethimide, cyproterone acetate, finasteride, buserelin acetate, fludrocortisone, fluoxyme sterone, medroxyprogesterone, octreotide), aromatase inhibitors (e.g. anastrozole, letrozole, liarozole, vorozole, exemestane, atamestane), LHRH agonists and antagonists (e.g. goserelin acetate, luprolide), inhibitors of growth factors and/or of their corresponding receptors (growth factors such as for example platelet derived growth factor (PDGF), fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), insuline-like growth factors (IGF), human epidermal growth factor (HER, e.g. HER2, HER3, HER4) and hepatocyte growth factor (HGF) and/or their corresponding receptors), inhibitors are for example (anti-)growth factor antibodies, (anti-)growth factor receptor antibodies and tyrosine kinase inhibitors, such as for example cetuximab, gefitinib, afatinib, nintedanib, imatinib, lapatinib, bosutinib, bevacizumab, pertuzumab and trastuzumab); antimetabolites (e.g. antifolates such as methotrexate, raltitrexed, pyrimidine analogues such as 5fluorouracil (5fluorineU), ribonucleoside and deoxyribonucleoside analogues, capecitabine and gemcitabine, purine and adenosine analogues such as mercaptopurine, thioguanine, cladribine and pentostatin, cytarabine (ara C), fludarabine); antitumor antibiotics (e.g. anthracyclins such as doxorubicin, doxil (pegylated liposomal doxorubicin hydrochloride, myocet (non-pegylated liposomal doxorubicin), daunorubicin, epirubicin and idarubicin, mitomycin-C, bleomycin, dactinomycin, plicamycin, streptozocin); platinum derivatives (e.g. cisplatin, oxaliplatin, carboplatin); alkylation agents (e.g. estramustin, meclorethamine, melphalan, chlorambucil, busulphan, dacarbazin, cyclophosphamide, ifosfamide, temozolomide, nitrosoureas such as for example carmustin and lomustin, thiotepa); antimitotic agents (e.g. Vinca alkaloids such as for example vinblastine, vindesin, vinorelbin and vincristine; and taxanes such as paclitaxel, docetaxel); angiogenesis inhibitors (e.g. tasquinimod), tubuline inhibitors; DNA synthesis inhibitors, PARP inhibitors, topoisomerase inhibitors (e.g. epipodophyllotoxins such as for example etoposide and etopophos, teniposide, amsacrin, topotecan, irinotecan, mitoxantrone), serine/threonine kinase inhibitors (e.g. PDK 1 inhibitors, Raf inhibitors, A-Raf inhibitors, B-Raf inhibitors, C-Raf inhibitors, mTOR inhibitors, mTORCl/2 inhibitors, PI3K inhibitors, PI3Ka inhibitors, dual mT0R/PI3K inhibitors, STK 33 inhibitors, AKT inhibitors, PLK 1 inhibitors, inhibitors of CDKs, Aurora kinase inhibitors), tyrosine kinase inhibitors (e.g. PTK2/FAK inhibitors), protein protein interaction inhibitors (e.g. IAP activator, Mcl-1, MDM2/MDMX), MEK inhibitors, ERK inhibitors, KRAS inhibitors (e.g. KRAS G12C inhibitors), signalling pathway inhibitors (e.g. SOS1 inhibitors), FLT3 inhibitors, BRD4 inhibitors, IGF-1R inhibitors, TRAILR2 agonists, Bcl-xL inhibitors, Bcl-2 inhibitors, Bcl-2/Bcl-xL inhibitors, ErbB receptor inhibitors, BCR-ABL inhibitors, ABL inhibitors, Src inhibitors, rapamycin analogs (e.g. everolimus, temsirolimus, ridaforolimus, sirolimus), androgen synthesis inhibitors, androgen receptor inhibitors, DNMT inhibitors, HDAC inhibitors, ANG1/2 inhibitors, CYP17 inhibitors, radiopharmaceuticals, proteasome inhibitors, immunotherapeutic agents such as immune checkpoint inhibitors (e.g. CTLA4, PD1, PD-L1, PD-L2, LAG3, and TIM3 binding molecules/immunoglobulins, such as e.g. ipilimumab, nivolumab, pembrolizumab), ADCC (antibody-dependent cell-mediated cytotoxicity) enhancers (e.g. anti-CD33 antibodies, anti-CD37 antibodies, anti-CD20 antibodies), T- cell engagers (e.g. bi-specific T-cell engagers (BiTEs®) like e.g. CD3 x BCMA, CD3 x CD33, CD3 x CD19), PSMA x CD3), tumor vaccines and various chemotherapeutic agents such as amifostin, anagrelid, clodronat, fdgrastin, interferon, interferon alpha, leucovorin, procarbazine, levamisole, mesna, mitotane, pamidronate and porfimer.

Cancer

In embodiments, the cancer is one of the following, without being restricted thereto: Cancers/tumors/carcinomas of the head and neck: e.g. tumors/carcinomas/cancers of the nasal cavity, paranasal sinuses, nasopharynx, oral cavity (including lip, gum, alveolar ridge, retromolar trigone, floor of mouth, tongue, hard palate, buccal mucosa), oropharynx (including base of tongue, tonsil, tonsillar pilar, soft palate, tonsillar fossa, pharyngeal wall), middle ear, larynx (including supraglottis, glottis, subglottis, vocal cords), hypopharynx, salivary glands (including minor salivary glands); cancers/tumors/carcinomas of the lung: e.g. non-small cell lung cancer (NSCLC) (squamous cell carcinoma, spindle cell carcinoma, adenocarcinoma, large cell carcinoma, clear cell carcinoma, bronchioalveolar), small cell lung cancer (SCLC) (oat cell cancer, intermediate cell cancer, combined oat cell cancer); neoplasms of the mediastinum: e.g. neurogenic tumors (including neurofibroma, neurilemoma, malignant schwannoma, neurosarcoma, ganglioneuroblastoma, ganglioneuroma, neuroblastoma, pheochromocytoma, paraganglioma), germ cell tumors (including seminoma, teratoma, nonseminoma), thymic tumors (including thymoma, thymolipoma, thymic carcinoma, thymic carcinoid), mesenchymal tumors (including fibroma, fibrosarcoma, lipoma, liposarcoma, myxoma, mesothelioma, leiomyoma, leiomyosarcoma, rhabdomyosarcoma, xanthogranuloma, mesenchymoma, hemangioma, hemangioendothelioma, hemangiopericytoma, lymphangioma, lymphangiopericytoma, lymphangiomyoma) ; cancers/tumors/carcinomas of the gastrointestinal (GI) tract: e.g. tumors/carcinomas/ cancers of the esophagus, stomach (gastric cancer), pancreas, liver and biliary tree (including hepatocellular carcinoma (HCC), e.g. childhood HCC, fibrolamellar HCC, combined HCC, spindle cell HCC, clear cell HCC, giant cell HCC, carcinosarcoma HCC, sclerosing HCC; hepatoblastoma; cholangiocarcinoma; cholangiocellular carcinoma; hepatic cystadenocarcinoma; angiosarcoma, hemangioendothelioma, leiomyosarcoma, malignant schwannoma, fibrosarcoma, Klatskin tumor), gall bladder, extrahepatic bile ducts, small intestine (including duodenum, jejunum, ileum), large intestine (including cecum, colon, rectum, anus; colorectal cancer, gastrointestinal stroma tumor (GIST)), appendix, genitourinary system (including kidney, e.g. renal pelvis, renal cell carcinoma (RCC), nephroblastoma (Wilms' tumor), hypernephroma, Grawitz tumor; ureter; urinary bladder, e.g. urachal cancer, urothelial cancer; urethra, e.g. distal, bulbomembranous, prostatic; prostate (androgen dependent, androgen independent, castration resistant, hormone independent, hormone refractory), penis); cancers/tumors/carcinomas of the testis: e.g. seminomas, non-seminomas;

Gynecologic cancers/tumors/carcinomas: e.g. tumors/carcinomas/cancers of the ovary, fallopian tube, peritoneum, cervix, vulva, vagina, uterine body (including endometrium, fundus); cancers/tumors/carcinomas of the breast: e.g. mammary carcinoma (infiltrating ductal, colloid, lobular invasive, tubular, adenocystic, papillary, medullary, mucinous), hormone receptor positive breast cancer (estrogen receptor positive breast cancer, progesterone receptor positive breast cancer), HER2 positive breast cancer, triple negative breast cancer, Paget's disease of the breast; cancers/tumors/carcinomas of the endocrine system: e.g. tumors/carcinomas/cancers of the endocrine glands, thyroid gland (thyroid carcinomas/tumors; papillary, follicular, anaplastic, medullary), parathyroid gland (parathyroid carcinoma/tumor), adrenal cortex (adrenal cortical carcinoma/tumors), pituitary gland (including prolactinoma, craniopharyngioma), thymus, adrenal glands, pineal gland, carotid body, islet cell tumors, paraganglion, pancreatic endocrine tumors (PET; nonfluorineunctional PET, PPoma, gastrinoma, insulinoma, VIPoma, glucagonoma, somatostatinoma, GRFoma, ACTHoma), carcinoid tumors; sarcomas of the soft tissues: e.g. fibrosarcoma, fibrous histiocytoma, liposarcoma, leiomyosarcoma, rhabdomyosarcoma, angiosarcoma, lymphangiosarcoma, Kaposi's sarcoma, glomus tumor, hemangiopericytoma, synovial sarcoma, giant cell tumor of tendon sheath, solitary fibrous tumor of pleura and peritoneum, diffuse mesothelioma, malignant peripheral nerve sheath tumor (MPNST), granular cell tumor, clear cell sarcoma, melanocytic schwannoma, plexosarcoma, neuroblastoma, ganglioneuroblastoma, neuroepithelioma, extraskeletal Ewing's sarcoma, paraganglioma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, mesenchymoma, alveolar soft part sarcoma, epithelioid sarcoma, extrarenal rhabdoid tumor, desmoplastic small cell tumor; sarcomas of the bone: e.g. myeloma, reticulum cell sarcoma, chondrosarcoma (including central, peripheral, clear cell, mesenchymal chondrosarcoma), osteosarcoma (including parosteal, periosteal, high-grade surface, small cell, radiation-induced osteosarcoma, Paget's sarcoma), Ewing's tumor, malignant giant cell tumor, adamantinoma, (fibrous) histiocytoma, fibrosarcoma, chordoma, small round cell sarcoma, hemangioendothelioma, hemangiopericytoma, osteochondroma, osteoid osteoma, osteoblastoma, eosinophilic granuloma, chondroblastoma; mesothelioma: e.g. pleural mesothelioma, peritoneal mesothelioma; cancers of the skin: e.g. basal cell carcinoma, squamous cell carcinoma, Merkel's cell carcinoma, melanoma (including cutaneous, superficial spreading, lentigo maligna, acral lentiginous, nodular, intraocular melanoma), actinic keratosis, eyelid cancer; neoplasms of the central nervous system and brain: e.g. astrocytoma (cerebral, cerebellar, diffuse, fibrillary, anaplastic, pilocytic, protoplasmic, gemistocytary), glioblastoma, gliomas, oligodendrogliomas, oligoastrocytomas, ependymomas, ependymoblastomas, choroid plexus tumors, medulloblastomas, meningiomas, schwannomas, hemangioblastomas, hemangiomas, hemangiopericytomas, neuromas, ganglioneuromas, neuroblastomas, retinoblastomas, neurinomas (e.g. acoustic), spinal axis tumors; peripheral nervous system cancer; lymphomas and leukemias: e.g. B-cell non-Hodgkin lymphomas (NHL) (including small lymphocytic lymphoma (SLL), lymphoplasmacytoid lymphoma (LPL), mantle cell lymphoma (MCL), follicular lymphoma (FL), diffuse large cell lymphoma (DLCL), Burkitt's lymphoma (BL)), T-cell non-Hodgkin lymphomas (including anaplastic large cell lymphoma (ALCL), adult T-cell leukemia/lymphoma (ATLL), cutaneous T-cell lymphoma (CTCL), peripheral T-cell lymphoma (PTCL)), lymphoblastic T- cell lymphoma (T-LBL), adult T-cell lymphoma, lymphoblastic B-cell lymphoma (B-LBL), immunocytoma, chronic B-cell lymphocytic leukemia (BchlorineL), chronic T-cell lymphocytic leukemia (TchlorineL) B-cell small lymphocytic lymphoma (B-SLL), cutaneous T-cell lymphoma (CTLC), primary central nervous system lymphoma (PCNSL), immunoblastoma, Hodgkin's disease (HD) (including nodular lymphocyte predominance HD (NLPHD), nodular sclerosis HD (NSHD), mixed-cellularity HD (MCHD), lymphocyte-rich classic HD, lymphocyte-depleted HD (LDHD)), large granular lymphocyte leukemia (LGL), chronic myelogenous leukemia (CML), acute myelogenous/myeloid leukemia (AML), acute lymphatic/lymphoblastic leukemia (ALL), acute promyelocytic leukemia (APL), chronic lymphocytic/lymphatic leukemia (CLL), prolymphocytic leukemia (PLL), hairy cell leukemia, chronic myelogenous/myeloid leukemia (CML), myeloma, plasmacytoma, multiple myeloma (MM), plasmacytoma, myelodysplastic syndromes (MDS), chronic myelomonocytic leukemia (CMML); cancers of unknown primary site (CUP).

All cancers/tumors/carcinomas mentioned above which are characterized by their specific location/origin in the body are meant to include both the primary tumors and the metastatic tumors derived therefrom. Preferably, the cancer as defined herein (including in any embodiment referring to e.g. cancer types) is metastatic, advanced, and/or unresectable.

All cancers/tumors/carcinomas mentioned above may be further differentiated by their histopathological classification:

Epithelial cancers, e.g. squamous cell carcinoma (SCC) (carcinoma in situ, superficially invasive, verrucous carcinoma, pseudosarcoma, anaplastic, transitional cell, lymphoepithelial), adenocarcinoma (AC) (well-differentiated, mucinous, papillary, pleomorphic giant cell, ductal, small cell, signet-ring cell, spindle cell, clear cell, oat cell, colloid, adenosquamous, mucoepidermoid, adenoid cystic), mucinous cystadenocarcinoma, acinar cell carcinoma, large cell carcinoma, small cell carcinoma, neuroendocrine tumors (small cell carcinoma, paraganglioma, carcinoid); oncocytic carcinoma;

Nonepithelial cancers, e.g. sarcomas (fibrosarcoma, chondrosarcoma, rhabdomyosarcoma, leiomyosarcoma, hemangiosarcoma, giant cell sarcoma, lymphosarcoma, fibrous histiocytoma, liposarcoma, angiosarcoma, lymphangiosarcoma, neurofibrosarcoma), lymphoma, melanoma, germ cell tumors, hematological neoplasms, mixed and undifferentiated carcinomas.

In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is manifested by at least one solid tumor.

In some embodiments, the cancer is selected from the group consisting of brain cancer, breast cancer, endocrine cancer, gastrointestinal cancer, gynecologic cancer, head and neck tumor, lung cancer, nervous system cancer, and skin cancer.

Preferably, said brain cancer is a glioblastoma or a glioma.

Preferably, said breast cancer is lobular breast cancer. In addition or in alternative, said breast cancer is preferably metastatic.

Preferably, said endocrine cancer is nerve sheath tumor, more preferably HER2 mutant nerve sheath tumor.

Preferably, said gastrointestinal cancer is selected from the group consisting of anal cancer, appendix cancer, biliary tract cancer, bladder cancer, colorectal cancer, esophagogastric cancer, gastric cancer, esophagus tumor, gastroesophageal cancer, gallbladder tumor, hepatobiliary cancer, kidney cancer, liver cancer, pancreatic cancer, prostate cancer and small bowel cancer. In addition or in alternative, said gastrointestinal cancer may be a gastrointestinal neuroendocrine tumor, preferably HER2 mutant. Still preferably, said gastrointestinal cancer is selected from the group consisting of gastric adenocarcinoma, gastroesophageal junction adenocarcinoma and esophageal adenocarcinoma, in particular metastatic gastric adenocarcinoma, metastatic gastroesophageal junction adenocarcinoma and metastatic esophageal adenocarcinoma.

Preferably, said gynecologic cancer is selected from the group consisting of cervical cancer, uterine cancer, endometrial cancer and ovarian cancer.

As used herein, “head and neck tumor” preferably refers to a head and neck cancer. Preferably, said head and neck tumor is a salivary gland cancer or tumor.

Preferably, said lung cancer is non-small cell lung cancer (NSCLC).

Preferably, said nervous system cancer is peripheral nervous system cancer, more preferably HER2 amplified peripheral nervous system cancer.

Preferably, said skin cancer is not a melanoma, i.e. non-melanoma skin cancer.

In some embodiments, the cancer is selected from the group consisting of glioblastoma, glioma, lobular breast cancer, metastatic breast cancer, nerve sheath tumor, anal cancer, appendix cancer, biliary tract cancer, bladder cancer, colorectal cancer, esophagogastric cancer, gastric cancer, esophagus tumor, gastroesophageal cancer, gallbladder tumor, hepatobiliary cancer, kidney cancer, liver cancer, pancreatic cancer, prostate cancer, small bowel cancer, neuroendocrine gastrointestinal cancer, metastatic gastric adenocarcinoma, metastatic gastroesophageal junction adenocarcinoma, metastatic esophageal adenocarcinoma, cervical cancer, uterine cancer, endometrial cancer, ovarian cancer, salivary gland cancer, non-small cell lung cancer (NSCLC), peripheral nervous system cancer and nonmelanoma skin cancer.

In some embodiments, the cancer is HER2 aberrant - preferably overexpressed, HER2 amplified and/or HER2 mutant (in particular HER2 exon 20 mutant) - cancer selected from the group consisting of glioblastoma, glioma, lobular breast cancer, metastatic breast cancer, nerve sheath tumor, anal cancer, appendix cancer, biliary tract cancer, bladder cancer, colorectal cancer, esophagogastric cancer, gastric cancer, esophagus tumor, gastroesophageal cancer, gallbladder tumor, hepatobiliary cancer, kidney cancer, liver cancer, pancreatic cancer, prostate cancer, small bowel cancer, neuroendocrine gastrointestinal cancer, metastatic gastric adenocarcinoma, metastatic gastroesophageal junction adenocarcinoma, metastatic esophageal adenocarcinoma, cervical cancer, uterine cancer, endometrial cancer, ovarian cancer, salivary gland cancer, non-small cell lung cancer (NSCLC), peripheral nervous system cancer and non-melanoma skin cancer.

In another aspect, the cancer is selected from the group consisting of brain cancer, breast cancer, biliary tract cancer, bladder cancer, cervical cancer, uterine cancer, colorectal cancer, endometrial cancer, ovarian cancer, skin cancer, gastric cancer, esophagus tumor, head and neck tumor, salivary gland cancer, gastrointestinal cancer, small bowel cancer, gallbladder tumor, kidney cancer, liver cancer, lung cancer and prostate cancer.

In some embodiments, the cancer is HER2 aberrant - preferably overexpressed, HER2 amplified and/or HER2 mutant (in particular HER2 exon 20 mutant) - cancer selected from the group consisting of brain cancer, breast cancer, biliary tract cancer, bladder cancer, cervical cancer, uterine cancer, colorectal cancer, endometrial cancer, ovarian cancer, skin cancer, gastric cancer, esophagus tumor, head and neck tumor, salivary gland cancer, gastrointestinal cancer, small bowel cancer, gallbladder tumor, kidney cancer, liver cancer, lung cancer and prostate cancer.

In other embodiments, the cancer is selected from the group consisting of breast cancer, bladder cancer, colorectal cancer, gastrointestinal cancer, esophageal cancer or lung cancer.

In further embodiments, the cancer is selected from cancers/tumors/carcinomas of the lung: e.g. nonsmall cell lung cancer (NSCLC) (squamous cell carcinoma, spindle cell carcinoma, adenocarcinoma, large cell carcinoma, clear cell carcinoma, bronchioalveolar), small cell lung cancer (SCLC) (oat cell cancer, intermediate cell cancer, combined oat cell cancer). In still further embodiments, the cancer is NSCLC. In still further embodiments, the cancer is HER2 exon 20 mutant NSCLC.

In another aspect, the cancer is selected from the group consisting of breast cancer, esophageal cancer, gastric cancer, gastroesophageal junction cancer, and lung cancer. Preferably, said lung cancer is nonsmall cell lung cancer (NSCLC).

In another aspect, said cancer is advanced, unresectable and/or metastatic.

In another aspect, said cancer is an adenocarcinoma.

In embodiments, the cancer or tumor comprises a HER2 aberration. This means that the cells of the cancer or tumor harbor an aberration of HER2. As used herein, the expressions “HER2 aberration”, “aberration of HER2” and grammatical variants thereof have the meaning commonly attributed to them in the art and include any variation or alteration in the HER2 protein or its encoding gene, such as: overexpression of the HER2 protein, amplification of the HER2 -encoding gene, mutations in the HER2- encoding gene and/or in the HER2 protein (in particular non-synonymous mutations, somatic mutations, mutations in specific regions, e.g. in the tyrosine kinase domain, in exon 20, etc.) as well as gene rearrangements of HER2 and/or NRG1. When the cancer comprises a HER2 aberration, it can be referred to as HER2 aberrant. When the cancer comprises an overexpression of the HER2 protein, it can be referred to as HER2 overexpressed. When the cancer comprises an amplification of the HER2- encoding gene, it can be referred to as HER2 amplified. When the cancer comprises a mutation in the HER2 -encoding gene and/or in the HER2 protein, it can be referred to as HER2 mutant.

In another aspect, the cancer is HER2 overexpressed, HER2 amplified and/or HER2 mutant.

In another aspect, the cancer is HER2 overexpressed and/or HER2 amplified.

In embodiments, the cancer comprises a mutation in the tyrosine kinase domain of HER2. In embodiments, the cancer is HER2 exon 20 mutant cancer. In embodiments, the cancer comprises a gene rearrangement of HER2 and/or NRG1.

The presence or absence of HER2 alterations, including overexpression, amplification or mutations, can be determined using methods known in the art.

“HER2 overexpressed” as used herein refers to a cancer comprising cells that express HER2 at levels detectable by immunohistochemistry (e.g. IHC 2+ and IHC 3+) and/or methods assaying ERBB2 messenger RNA.

“HER2 amplified” as used herein refers to a cancer comprising cells exhibiting more than 2, in particular more than 3, 4, 5, 6, 7, 8, 9 or 10, preferably more than 6, copies of the HER2 gene ERBB2.

HER2 expression, gene copy number or amplification can be measured, for example, by determining nucleic acid sequencing (e.g., sequencing of genomic DNA or cDNA), measuring mRNA expression, measuring protein abundance, or a combination thereof. HER2 testing methods include immunohistochemistry (IHC), in situ hybridization - including fluorescence in situ hybridization (FISH) and chromogenic in situ hybridization (CISH) - ELISAs, and RNA quantification using techniques such as Reverse Transcription- Polymerase Chain Reaction (RT-PCR), microarray analysis and Next Generation Sequencing (NGS). HER2 expression can be compared to a reference cell. The reference cell can be a non-cancer cell obtained from the same subject as the sample cell. The reference cell can be a non-cancer cell obtained from a different subject or a population of subjects.

When the cancer is HER2 overexpressed and/or HER2 amplified in or on a cell, the cancer can be referred to as being “HER2 positive”. The level of HER2 amplification or overexpression in HER2 positive cancers is commonly expressed as a score ranging from 0 to 3 (i.e., HER2 0, HER2 1+, HER2 2+, or HER2 3+), with higher scores corresponding to greater degrees of expression.

In an aspect, the cancer is HER2 positive.

Preferably, “HER2 overexpressed”, “HER2 amplified” and “HER2 positive” mean that the cancer comprises cells having an immunohistochemistry score of 2+ or 3+.

Preferably, “HER2 overexpressed”, “HER2 amplified” and “HER2 positive” mean that the cancer comprises cells having HER2 amplification defined by in situ hybridization.

In some embodiments, the HER2 status of the cancer, specifically of a sample cell within the cancer, is determined. The determination can be made before the combination treatment begins, during treatment, or after treatment has been completed. In some instances, determination of the HER2 status results in a decision to change therapy.

The sample cell can be obtained as a biopsy specimen, by surgical resection, or as a fine needle aspirate (FNA).

In some embodiments, the sample cell is determined to be HER2 positive when HER2 is expressed at a higher level in the sample cell compared to a reference cell. In some embodiments, the cell is determined to be HER2 positive when HER2 is overexpressed at least about 1.5-fold (e.g., about 1.5- fold, 2-fold, 2.5-fold, 3-fold, 3.5-fold, 4-fold, 4.5-fold, 5- fold, 5.5-fold, 6-fold, 6.5-fold, 7-fold, 7.5- fold, 8-fold, 8.5-fold, 9-fold, 9.5-fold, 10-fold, 1 1- fold, 12-fold, 13-fold, 14-fold, 15-fold, 16-fold, 17- fold, 18-fold, 19-fold, 20-fold, 25-fold, 30-fold, 35-fold, 40-fold, 45-fold, 50-fold, 55-fold, 60-fold, 65- fold, 70-fold, 75-fold, 80- fold, 85-fold, 90-fold, 95-fold, 100-fold, or more) compared to a reference cell. In particular embodiments, the cell is determined to be HER2 positive when HER2 is overexpressed at least about 1.5-fold compared to the reference cell.

In some embodiments, the sample cell is determined to be HER2 positive when the FISH or CISH signal ratio is greater than 2.

“HER2 mutant” as used herein refers to a cancer harbouring at least one mutation, i.e. an alteration in the nucleic acid sequence of the HER2 gene and/or an alteration in the amino acid sequence of the HER2 protein, including but not limited to those listed below. Mutations can be found with any method known to the skilled person, such as molecular diagnostic methods including but not limited to Polymerase Chain Reaction (PCR), Single Strand Conformational Polymorphism (SSCP), Denaturing Gradient Gel Electrophoresis (DGGE), Heteroduplex analysis, Restriction fragment length polymorphism (RFLP), Next Generation Sequencing and Whole Exome Sequencing.

In embodiments of said HER2 mutant cancer, the mutation is a non-synonymous mutation. As used herein, the term “non-synonymous” has the meaning commonly attributed to it in the art and in particular refers to a mutation in the nucleic acid sequence of the HER2 -encoding gene that alters the amino acid sequence of the HER2 protein.

In embodiments of said HER2 mutant cancer, the mutation is a somatic mutation. As used herein, the term “somatic” has the meaning commonly attributed to it in the art and in particular refers to a mutation in the nucleic acid sequence of the HER2 -encoding gene occurring in a cell other than a gamete, a germ cell or a gametocyte.

In embodiments of said HER2 mutant cancer, the mutation is a non-synonymous somatic mutation.

In embodiments of said HER2 mutant cancer, the mutation is a non-synonymous somatic mutation in the tyrosine kinase domain of HER2.

In embodiments of said HER2 mutant cancer, the mutation is in the tyrosine kinase domain of HER2, in particular in the exon 20 of HER2. In the latter case, the cancer can be referred to as HER2 exon 20 mutant.

As used herein, “HER2 mutant” may also refer to a rearrangement involving the HER2 gene ERBB2 and/or the NRG1 gene.

As used herein, a cancer comprising a mutation in the tyrosine kinase domain of HER2 is a cancer where the cancer or tumor cells harbour at least one mutation in the tyrosine kinase domain of HER2, which ranges from amino acids 694 to 883 and/or exons 18 to 21.

“Cancer with HER2 exon 20 mutation” or “HER2 exon 20 mutant cancer” as used herein refers to a cancer where the cancer or tumor cells harbour at least one HER2 exon 20 mutation including but not limited to the mutations listed below. ERBB2 (HER2) exon 20 encodes for a part of the kinase domain and ranges from amino acids 769 to 835. Every mutation, insertion, duplication or deletion within this region is defined as an exon 20 mutation including the following mutations: p.A772_G773insMMAY; p.Y772_A775_dup (YVMA); p.A775_G776insYVMA; p.Y772insYVMA; p.M774delinsWLV; p.A775_G776insSVMA; p.A775_G776insVVMA; p.A775_G776insYVMS; p.A775_G776insC; p.A776_delinsVC; p.A776_delinsLC; p.A776_delinsVV; p.A776_delinsAVGC; p.A776_delinsIC; p.A776_V777delinsCVC; p.V777_insE; p.V777_G778insV; p.V777_G778insC; p.V777_G778insCG; p.V777_S779dup; p.V777L; p.V777M; p.G778_P780dup (GSP); p.G778_S779insCPG; p.G778_S779insG; p.G776_delinsVC; p.G776_V777delinsAVGCV; p.G776delinsLC; p.G776_V777delinsAVCV; p.G776delinsVV; p.G776_V777insL; p.G776_V777insVGC; p.G776C; p.G776A; p.G776L; p.G776V; p.P780_Y781insGSP (“p.” is referring to the HER2 protein).

In addition HER2 mutations exist outside of exon 20 including the following mutations: p.S310A; p.S310F; p.S310Y; p.R678Q; p.G727A; p.T733I; p.L755S; p.L755A; p.L755F; p.L755P; p.L755S; p.V842I; p.D769Y; p.D769H; p.R103Q; p.G1056S; p.I767M; p.L869R; p.L869R; p.T733I; p.T862A; p.V697L; p.R929W; p.D277H; p.D277Y; p.G660D (“p.” is referring to the HER2 protein).

Of these, examples of tyrosine kinase mutations include: p.G727A; p.T733I; p.L755S; p.L755A; p.L755F; p.L755P; p.L755S; p.V842I; p.D769Y; p.D769H; p.I767M; p.L869R; p.L869R; p.T733I; p.T862A; p.V697L.

In an aspect, the cancer is HER2 positive breast cancer, in particular advanced HER2 positive breast cancer or HER2 positive metastatic breast cancer, preferably advanced HER2 positive metastatic breast cancer. Preferably, in this embodiment, the combination as described herein is administered as first line of therapy. Still preferably, in this embodiment, the combination as described herein is administered as second or further line of therapy.

In an aspect, the cancer is HER2 positive esophageal cancer, HER2 positive gastric cancer, or HER2 positive gastroesophageal junction cancer, in particular HER2 positive esophageal adenocarcinoma, HER2 positive gastric adenocarcinoma, or HER2 positive gastroesophageal junction adenocarcinoma, preferably metastatic HER2 positive esophageal adenocarcinoma, metastatic HER2 positive gastric adenocarcinoma, or metastatic HER2 positive gastroesophageal junction adenocarcinoma. Preferably, in this embodiment, the combination as described herein is administered as first line of therapy. Still preferably, in this embodiment, the combination as described herein is administered as second or further line of therapy.

In an aspect, the cancer is HER2 mutant lung cancer, in particular HER2 exon 20 mutant lung cancer or HER2 mutant NSCLC, preferably HER2 exon 20 mutant NSCLC.

In an embodiment, the cancer is advanced, unresectable or metastatic NSCLC harbouring a HER2 mutation, wherein said HER2 mutation is in the tyrosine kinase domain. Preferably, in this embodiment, the combination as described herein is administered as first line of therapy. Still preferably, in this embodiment, the combination as described herein is administered as second or further line of therapy.

Pharmaceutical composition and kit

It is provided a pharmaceutical composition comprising zongertinib or a pharmaceutically acceptable salt thereof, a SOS1 inhibitor and a pharmaceutically acceptable excipient. Preferably, said pharmaceutical composition is for use in the treatment and/or prevention of cancer.

The term “pharmaceutically acceptable excipient” refers to a non-toxic component that does not destroy the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable excipients that may be used in the compositions of this invention include fillers, disintegrants, glidants, lubricants, and coating agents. The compositions may comprise further pharmaceutically acceptable excipients selected from buffers, dispersion agents, surfactants, wetting agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, processing aids, colorants, sweeteners, perfuming agents, flavoring agents, diluents and other known additives usable in the manufacturing of a pharmaceutical product. Pharmaceutical compositions as referred to herein may contain conventional non-toxic pharmaceutically acceptable excipients.

Also provided herein is a pharmaceutical composition comprising zongertinib or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient, for use in the treatment and/or prevention of cancer, wherein the pharmaceutical composition is administered in combination with a SOS1 inhibitor.

The terms “first” and “second” with respect to pharmaceutical compositions, as used herein, are solely intended to indicate that these compositions are two different compositions. Thus, these terms shall not be understood to refer to the order or sequence of administration.

In an embodiment, the kit as defined herein additionally comprises a package insert. Preferably, the package insert comprises instructions. Still preferably, the instructions provide guidance on simultaneous, concurrent, sequential, successive, alternate or separate administration of zongertinib, its pharmaceutically acceptable salt or pharmaceutical composition and the SOS1 inhibitor.

In an embodiment, the kit is for use as a medicament.

In an embodiment, the kit is for use in the treatment and/or prevention of cancer.

Also provided herein is a method of treating and/or preventing cancer, the method comprising administering to a patient in need thereof a kit comprising:

(i) a first pharmaceutical composition comprising zongertinib or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient; and

(ii) a second pharmaceutical composition comprising a SO SI inhibitor and a pharmaceutically acceptable excipient. In an embodiment, the first pharmaceutical composition comprises a therapeutically effective amount of zongertinib or the pharmaceutically acceptable salt thereof and the second pharmaceutical composition comprises a therapeutically effective amount of the SOS1 inhibitor.

Also provided herein is a use of the kit as defined herein for the manufacture of a medicament for the treatment and/or prevention of cancer.

Also provided herein is a method of treating and/or preventing cancer, the method comprising administering to a patient in need thereof a pharmaceutical composition comprising: (i) zongertinib or a pharmaceutically acceptable salt thereof, (ii) a SOS1 inhibitor and (iii) a pharmaceutically acceptable excipient.

In an embodiment, said pharmaceutical composition comprises a therapeutically effective amount of zongertinib or the pharmaceutically acceptable salt thereof and a therapeutically effective amount of the SOS 1 inhibitor.

Also provided herein is a use of the pharmaceutical composition as defined herein for the manufacture of a medicament for the treatment and/or prevention of cancer.

Features and advantages of the present invention will become apparent from the following detailed examples, which illustrate the principles of the invention by way of example without restricting its scope:

Preparation of the SOS1 inhibitors

The compound of formula (I) and intermediates are prepared by the methods of synthesis described hereinafter in which the substituents of the general formulae have the meanings given hereinbefore. These methods are intended as an illustration of the invention without restricting its subject matter and the scope of the compounds claimed to these examples. Where the preparation of starting compounds is not described, they are commercially obtainable or their synthesis is described in the prior art or they may be prepared analogously to known prior art compounds or methods described herein, i. e. it is within the skills of an organic chemist to synthesize these compounds. Substances described in the literature can be prepared according to the published methods of synthesis.

The general processes for preparing the compounds according to the formula (I) will become apparent to the one skilled in the art studying the following schemes. Starting materials may be either commercially available or may be prepared by methods that are described in the literature or herein or may be prepared in an analogous or similar manner. Any functional groups in the starting materials or intermediates may be protected using conventional protecting groups. These protecting groups may be cleaved again at a suitable stage within the reaction sequence using methods familiar to the one skilled in the art.

The scheme below (Scheme 1) illustrates the possible synthesis of the compounds of general formula (I) and its intermediates. The first step can be an amide coupling using carboxylic acid derivatives of structure B and benzylic amines of structure A as reaction partners to form bicyclic intermediates of structure C containing an aromatic bicyclic 5-6 fused ring system with an attached functional group 1 (FG1). The functional group 1 enables use of cross coupling reactions such as Suzuki reactions, Buchwald couplings, or alike in the next reaction step the. This allows either to form directly compounds of general formula (I) or the intermediate structures D. The functional group 2 (FG2) in intermediate D can be further derivatized using for example reductive amination reactions, nucleophilic substitution reactions or alike with reagents known to the people skilled in the art to form compounds of general formula (I). Scheme 1: Schematic synthesis of compounds of general formula (I).

As described for some exemplified compounds, a reaction called Dimroth rearrangement (described for example herein: ACS Med. Chem. Lett. 2017, 8, 12, 1320-1325) can also be used to form intermediates of structure C, which can be used in the synthesis of compound of general formula (I).

Abbreviations

Experimental part - Chemical synthesis

Other features and advantages of the present invention will become apparent from the following more detailed examples which exemplarily illustrate the principles of the invention without restricting its scope.

General

The terms "cis" and "trans" are used in accordance with the IUPAC Gold Book's guidelines to denote the stereochemical information of the substituents, which varies based on the positions of atoms (or groups) relative to a reference plane in a ring system. In the cis-isomer, the atoms are located on the same side, while in the trans-isomer, they are on opposing sides. This follows the traditional order of priority for substituents/ligands. (PAC, 1996, 68, 2193. (Basic terminology of stereochemistry (IUPAC Recommendations 1996)) on page 2203).

Unless stated otherwise, all the reactions are carried out in commercially obtainable apparatuses using methods that are commonly used in chemical laboratories. Starting materials that are sensitive to air and/or moisture are stored under protective gas and corresponding reactions and manipulations therewith are carried out under inert gas (nitrogen or argon).

Room temperature in the following schemes means the temperature ranging from 19 °C to 24 °C.

If a compound is to be represented both by a structural formula and by its nomenclature, in the event of a conflict the structural formula is decisive.

Some compounds according to the exemplified preparation are filtered through carbonate functionalized MP resin (carbonate cartridge) by Agilent Technologies (PU-HCOs MP SPE; Part. No. PE3540-C603) as indicated.

Some compounds according to the exemplified preparation are filtered through thiol functionalized SPEmedia (thiol resin) by Agilent Technologies (PL-Thiol MP SPE Part. No. 3582-CM89) prior to chromatography, if indicated.

The following catalyst, termed catalyst I, is used for some exemplified coupling reactions of this invention [ 1 , 3 -bis [2, 6-bis( 1 -ethylpropyl)phenyl]-4,5-dichloro-imidazol-2-yl]-dichloro-(2 -methyl- 1 - pyridyl)palladium (catalyst I; CAS: 1612891-29-8).

The following catalyst, termed catalyst II, is used for some exemplified coupling reactions of this invention [l,T-bis(diphenylphosphino)ferrocene-palladium(II)dichloride dichloromethane complex (CAS: 95464-05-4).

NMR method

NMR spectra were recorded on a Bruker AVANCE IIIHD 400 MHz instrument using TopSpin 3.2 pl6 software. Chemical shifts are given in parts per million (ppm) downfield from an internal reference like trimethylsilane and/or water and/or solvent (eg. d⁶-DMSO) in 5 units. Selected data are reported in the following manner: chemical shift (number of hydrogens). Chromatography

Thin layer chromatography is carried out on ready-made TLC plates of silica gel 60 on glass (with fluorescence indicator F-254) made by Merck.

Prep. RP-HPLC is carried out with columns made by Waters (Sunfire C18, 10 pm, 30x100 mm Part. No. 186003971 or X-Bridge C18, 10 pm, 30x100 mm Part. No. 186003930). The compounds are eluted using either different gradients of H2O/ACN or FFO/McOH. where 0.1% TFA is added to the water, or with different gradients utilizing a basic aqueous buffer solution (I L water contains 5 mL of an ammonium hydrogencarbonate solution (158 g per 1 L H2O) and 2 mL NH3 (7 mol/L solution in MeOH)) instead of the water-TFA-mixture. Mass spectroscopy

Low resolution mass spectra are obtained using a high performance liquid chromatography coupled to a quadrupole mass spectrometer (HPLC-MS; electrospray positive ionization). The reported mass spectrometry (MS) data correspond to the observed mass peaks of the monoisotopic masses of the respective compound ([M+H]⁺) or a fragment thereof (e.g., [M-Boc+H ]⁺, [M-NH3+H ]⁺). Analytical HPLC Methods (A.M. ) 12-0519-WO-1 Mobile phase preparations 5 Examples: The mobile phase “Water 0.1% TFA (v/v)” is prepared by adding 1 mL of a commercially available TFA solution to 999 mL water. The mobile phase “Water 0.1% NH3” is prepared by adding 4 mL of a commercially available concentrated ammonium hydroxide solution (25 wt%) to 996 mL water. 10 63 Analytical SFC Methods (A.M.)

Mobile phase preparations

The mobile phase “MEOH 20 mM NH3” is prepared by adding 3 ml of a commercially available solution of ammonia (7 M in methanol) to 997 ml methanol. The mobile phase “IP A 20 mM NH3” is prepared by adding 3 ml of a commercially available solution of ammonia (7 M in methanol) to 997 ml isopropyl alcohol.

The mobile phase “ETOH 20 mM NH3” is prepared by adding 3 ml of a commercially available solution of ammonia (7 M in methanol) to 997 ml ethanol.

Preparations

Synthesis scheme of intermediate 9 Methyl 6-amino-5 -bromonicotinate 1 (14.8 g, 61 mmol) and A. '-dimcthylfonnamidc dimethyl acetal (17.2 mL, 122 mmol) in DMF (141 mL) is stirred for 4 h at 100 °C. The solvents are evaporated under reduced pressure to obtain intermediate 2. The intermediate is used without further purification.

Synthesis of intermediate 3

Intermediate 2 (17.5 g, 61 mmol), sodium acetate (11 g, 134 mmol) and hydroxylamine hydrochloride (6.4 g, 91 mmol) in ethanol (249 mL) are stirred for 3 h at 50 °C. The mixture is poured into water and is further stirred for 1 h at rt. The solids are filtered off, washed with water and dried in vacuo at 55 °C to obtain intermediate 3. The intermediate is used without further purification.

Intermediate 3

Analytical HPLC-MS Method: D

Rt [min]: 0.83 MS [m/z]: 274 [M+H]⁺

Synthesis of intermediate 4

Intermediate 3 (16.7 g, 61 mmol) in THF (201 mL) is cooled to 0 °C. Trifluoroacetic anhydride (11 mL, 79 μmol) is added dropwise. After complete addition, the mixture is allowed to reach rt and stirring is continued for 16 h. The mixture is basified with NaHCOs solution under ice cooling, diluted with water and stirred for 2 h. The solvent is reduced under reduced pressure and the precipitate is filtered off, washed with water and dried in vacuo at 55 °C to obtain intermediate 4. The intermediate 4 is used without further purification.

Intermediate 4

Analytical HPLC-MS Method: D

Rt [min]: 0.75 MS [m/z]: 256 [M+H]⁺

An aqueous NaOH solution (4 M; 14.5 mL, 68 mmol) is added to intermediate 4 (13.5 g, 53 mmol) in MeOH (149 mL) and stirred for 1 h at 50 °C. The mixture is further stirred at rt for 16 h. MeOH is evaporated under reduced pressure before the mixture is diluted with water and fdtered. The aqueous mixture is acidified with aqueous 4 M HC1 solution. The mixture is stirred for 1 h. The precipitate is filtered off, washed with water and dried in vacuo at 55 °C to obtain intermediate 5. The intermediate 5 is used without further purification.

Intermediate 5

Analytical HPLC-MS Method: E

Rt [min] : 0.66 MS [m/z] : 242 [M+H]⁺

Synthesis of intermediate 7

Intermediate 5 (140 mg, 578 μmol) in DMF (9.4 mL) is treated with HATU (330 mg, 868 μmol) and triethylamine (325 pL, 2.3 mmol) and stirred for 15 h at rt. Benzylamine 6 (CAS: 1389852-29-2; 131 mg, 578 μmol) is added and the mixture is stirred for 16 h. MeOH/THF is added, solids are filtered off and the mixture is purified by prep. RP-HPLC (acidic conditions) to obtain intermediate 7.

Intermediate 7

Analytical HPLC-MS Method: D

Rt [min] : 0.94 MS [m/z] : 413 [M+H]⁺ 12-0519-WO-1 Synthesis of intermediate 9 A_{mixture of intermediate 7 (7.0 g, 16 mmol), piperidin-4-one hydrochloride 8 (2.5 g, 18 mmol), K3PO4} 5 (10.4 g, 48 mmol), and catalyst I (406 mg, 483 µmol) in 1,4-dioxane (70 mL) is stirred under argon for 16 h at 110 °C. The mixture is diluted with EtOAc and filtered. The solvent is removed under reduced pressure. The material is taken up in EtOAc and extracted with water and saturated aqueous NaCl solution. The separated organic layer is dried over Na2SO4, filtered and the solvent is evaporated under reduced pressure. The material is purified by column chromatography (SiO2; EtOAc/petroleum ether:10 1:3 à 1:0) to obtain intermediate 9. Intermediate 9 Analytical HPLC-MS Method: D R_t [min]: 0.91 MS [m/z]: 432 [M+H]⁺ 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.48 - 1.57 (3 H), 2.51 - 2.57 (4 H), 3.97 - 4.07 (4 H),} _{5.37 - 5.47 (1 H), 7.07 - 7.38 (3 H), 7.49 - 7.57 (1 H), 7.65 - 7.74 (1 H), 8.50 - 8.59 (1 H), 9.05 -} 9.14 (2 H). 77 Synthesis of intermediate 11

10 11

Hydrazine hydrate (45 ml, 603 mmol, 64-65%) is added to a mixture of 5-bromo-6-chloronicotinic acid 10 (50 g, 201 mmol) in ethanol (410 mL). The mixture is refluxed for 16 h. After cooling to rt, the solids are filtered off, washed with ethanol, and dried in vacuo to obtain intermediate 11. The material is used without further purification.

Intermediate 11

Analytical HPLC-MS Method: E

Rt [min]: 0.16 MS [m/z]: 232 [M+H]⁺

Synthesis of intermediate 12

Intermediate 11 (16.9 g, 73 mmol) is treated with formic acid (40 mL, 1.1 mmol) and stirred at 100 °C for 2 h. The mixture is cooled to 0 °C. The solid material is filtered off and dried under reduced pressure to obtain the intermediate 12. The material is used without further purification.

Intermediate 12

Analytical HPLC-MS Method: E

Rt [min] : 0.56 MS [m/z] : 242 [M+H]⁺

Synthesis of intermediate 13

A mixture of intermediate 12 (6.3 g, 25 mmol), benzylamine 6 (CAS: 1389852-29-2; 6.0 g, 26 mmol) and 4-methyhnorpholine (11 mL, 99 mmol) in ACN (60 mL) is cooled down to 0 °C. 1- Propanephosphonic acid anhydride (37 mL, 62 mmol) is added dropwise. The mixture is allowed to reach rt before it is poured into water and stirred for 20 min. The solids are filtered and washed with 12-0519-WO-1 A_{CN/water (1:1). The material is dried in vacuo (55 °C) to obtain intermediate 13. The material is used} without further purification. Intermediate 13 Analytical HPLC-MS Method: E R_{[min]: 0.92 MS [m/z] +} _{t : 413 [M+H]} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.48 - 1.72 (9 H), 1.85 - 1.94 (2 H), 2.43 - 2.48 (1 H),} _{2.62 - 2.72 (1 H), 2.80 - 2.92 (4 H), 3.02 - 3.10 (1 H), 4.28 - 4.41 (3 H), 5.36 - 5.46 (1 H), 7.07 -} _{7.37 (3 H), 7.49 - 7.56 (1 H), 7.65 - 7.72 (1 H), 8.47 - 8.53 (1 H), 9.02 - 9.10 (2 H), missing} proton(s) presumably hidden by/ overlapping with solvent signals. 5 Synthesis of intermediate 7 1_{3 7} _{A mixture of intermediate 13 (710 mg, 1.7 mmol), Cs2CO3 (280 mg, 859 µmol), piperidine (85 µL, 859} µmol) in 1,4-dioxane (10 mL) is stirred 6 h at 100 °C. EtOAc and water are added. The aqueous layer 10 is separated and extracted with EtOAc. The combined organic layers are dried over Na2SO4. The solids are filtered, and the solvent is evaporated under reduced pressure. The material is purified by column chromatography (SiO2; EtOAc/cyclohexane: 50:50 à 100:0) to obtain intermediate 7. I_{ntermediate 7} Analytical HPLC-MS Method: D Rt [min]: 0.94 MS [m/z]: 413 [M+H]⁺ 15 79 12-0519-WO-1 A_{mixture of Intermediate 9 (34.5 mg, 80 µmol), D-prolinol 14 (12.1 mg, 120 µmol), AcOH (30.0 µL,} 51 µmol), and 2-picoline-borane complex (8.6 mg, 80 µmol) in MeOH (2.0 mL) is stirred for 12 h at rt. T_{he mixture was diluted with DMF und purified by prep. RP-HPLC (basic conditions) to obtain the} example E1. Example E1 HPLC-MS Method: A R [min]: 0.73 MS [ + t m/z]: 517 [M+H] Analytical SFC method: AU R_{t [min]: 1.66 d.e. > 95%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.48 - 1.72 (9 H), 1.85 - 1.94 (2 H), 2.43 - 2.48 (1 H),} _{2.62 - 2.72 (1 H), 2.80 - 2.92 (4 H), 3.02 - 3.10 (1 H), 4.28 - 4.41 (3 H), 5.36 - 5.46 (1 H), 7.07 -} _{7.37 (3 H), 7.49 - 7.56 (1 H), 7.65 - 7.72 (1 H), 8.47 - 8.53 (1 H), 9.02 - 9.10 (2 H), missing} proton(s) presumably hidden by/ overlapping with solvent signals. 5 E_{xample E2 is prepared in analogy to example E1: Intermediate 9 (80 µmol), (R)-3-hydroxypyrrolidine} _{10 hydrochloride 15 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane complex (80 µmol), 2 mL} MeOH. Example E2 Analytical HPLC-MS Method: D Rt [min]: 0.91 MS [m/z]: 503 [M+H]⁺ Analytical SFC method: L R_{t [min]: 6.60 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.47 - 1.61 (6 H), 1.88 - 2.02 (3 H), 2.17 - 2.27 (1 H),} _{2.35 - 2.42 (1 H), 2.60 - 2.69 (1 H), 2.74 - 2.83 (1 H), 2.92 - 3.03 (2 H), 4.13 - 4.25 (3 H), 4.58 -} _{4.70 (1 H), 5.36 - 5.46 (1 H), 7.06 - 7.38 (3 H), 7.49 - 7.56 (1 H), 7.65 - 7.73 (1 H), 8.50 (1 H),} _{9.01 - 9.11 (2 H), missing proton(s) presumably hidden by/ overlapping with solvent signals.} 80 12-0519-WO-1 5_{Example E3 is prepared in analogy to example E1: Intermediate 9 (80 µmol), (7R)-5-} _{azaspiro[2.4]heptan-7-ol 16 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane complex (80} µmol), 2 mL MeOH. Example E3 Analytical HPLC-MS Method: A R_t [min]: 0.76 MS [m/z]: 529 [M+H]⁺ Analytical SFC method: L R_{t [min]: 2.26 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.30 - 0.40 (1 H), 0.45 - 0.58 (2 H), 0.77 - 0.86 (1 H),} _{1.45 - 1.61 (5 H), 1.87 - 1.98 (2 H), 2.18 - 2.27 (1 H), 2.41 - 2.48 (1 H), 2.60 - 2.66 (1 H), 2.93 -} _{3.04 (2 H), 3.04 - 3.12 (1 H), 3.68 - 3.77 (1 H), 4.16 - 4.26 (2 H), 4.47 - 4.56 (1 H), 5.36 - 5.46 (1} _{H), 7.07 - 7.38 (3 H), 7.49 - 7.56 (1 H), 7.64 - 7.72 (1 H), 8.47 - 8.52 (1 H), 9.02 - 9.11 (2 H),} missing proton(s) presumably hidden by/ overlapping with solvent signals. 10 E_{xample E4 is prepared in analogy to example E1: Intermediate 9 (80 µmol), (3S)-3-methylpyrrolidin-} _{3-ol hydrochloride 17 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane complex (80 µmol), 2} mL MeOH. Example E4 Analytical HPLC-MS Method: A R_t [min]: 0.75 MS [m/z]: 517 [M+H]⁺ 81 12-0519-WO-1 Analytical SFC method: F R_{t [min]: 1.56 d.e. > 90%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.21 - 1.27 (3 H), 1.48 - 1.61 (5 H), 1.62 - 1.77 (2 H),} _{1.87 - 1.98 (2 H), 2.18 - 2.29 (1 H), 2.52 - 2.61 (3 H), 2.65 - 2.74 (1 H), 2.94 - 3.05 (2 H), 4.14 -} _{4.25 (2 H), 4.41 - 4.50 (1 H), 5.36 - 5.46 (1 H), 7.07 - 7.38 (3 H), 7.48 - 7.57 (1 H), 7.64 - 7.72 (1} _{H), 8.48 - 8.53 (1 H), 9.02 - 9.10 (2 H).} 5_{Example E5 is prepared in analogy to example E1: Intermediate 9 (80 µmol), (3R)-3-methylpyrrolidin-} _{3-ol hydrochloride 18 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane complex (80 µmol), 2} mL MeOH. Example E5 Analytical HPLC-MS Method: A R [min]: 0. + t 75 MS [m/z]: 517 [M+H] Analytical SFC method: L R_{t [min]: 2.02 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.19 - 1.28 (3 H), 1.46 - 1.60 (5 H), 1.62 - 1.77 (2 H),} _{1.87 - 1.97 (2 H), 2.19 - 2.29 (1 H), 2.52 - 2.62 (3 H), 2.65 - 2.74 (1 H), 2.95 - 3.05 (2 H), 4.14 -} _{4.26 (2 H), 4.41 - 4.49 (1 H), 5.36 - 5.46 (1 H), 7.06 - 7.38 (3 H), 7.48 - 7.56 (1 H), 7.64 - 7.72 (1} _{H), 8.47 - 8.52 (1 H), 9.02 - 9.12 (2 H).} 10 82 12-0519-WO-1 E_{xample E6 is prepared in analogy to example E1: Intermediate 9 (80 µmol), L-prolinol 19 (0.12} mmol), AcOH (0.51 mmol), and 2-picoline-borane complex (80 µmol), 2 mL MeOH. Example E6 Analytical HPLC-MS Method: A Rt [min]: 0.76 MS [m/z]: 517 [M+H]⁺ Analytical SFC method: AP R_{t [min]: 1.91 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.48 - 1.72 (9 H), 1.85 - 1.94 (2 H), 2.62 - 2.71 (1 H),} _{2.79 - 2.93 (4 H), 3.01 - 3.11 (1 H), 4.27 - 4.41 (3 H), 5.36 - 5.47 (1 H), 7.07 - 7.37 (3 H), 7.49 -} _{7.56 (1 H), 7.65 - 7.72 (1 H), 8.48 - 8.53 (1 H), 9.02 - 9.11 (2 H), missing proton(s) presumably} hidden by/ overlapping with solvent signals. Synthesis of example E7 5 E_{xample E7 is prepared in analogy to example E1: Intermediate 9 (80 µmol), 2-methyl-1-} _{(methylamino)propan-2-ol 20 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane complex (80} µmol), 2 mL MeOH. Example E7 Analytical HPLC-MS Method: A Rt [min]: 0.84 MS [m/z]: 519 [M+H]⁺ Analytical SFC method: AR R_{t [min]: 5.33 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.04 - 1.11 (6 H), 1.49 - 1.54 (3 H), 1.55 - 1.65 (2 H),} _{1.77 - 1.86 (2 H), 2.26 - 2.31 (2 H), 2.31 - 2.35 (3 H), 2.53 - 2.62 (1 H), 2.73 - 2.83 (2 H), 3.92 -} _{3.99 (1 H), 4.36 - 4.46 (2 H), 5.36 - 5.46 (1 H), 7.08 - 7.38 (3 H), 7.49 - 7.56 (1 H), 7.64 - 7.72 (1} _{H), 8.49 - 8.53 (1 H), 9.02 - 9.11 (2 H).} 10 83 12-0519-WO-1 Synthesis of example E8 E_{xample E8 is prepared in analogy to example E1: Intermediate 9 (80 µmol), 5-azaspiro[2.4]heptane} _{5 hydrochloride 21 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane complex (80 µmol), 2 mL} MeOH. Example E8 Analytical HPLC-MS Method: A R_t [min]: 0.94 MS [m/z]: 513 [M+H]+ Analytical SFC method: AF R_{t [min]: 0.98 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.44 - 0.55 (4 H), 1.49 - 1.62 (5 H), 1.68 - 1.75 (2 H),} _{1.89 - 1.98 (2 H), 2.17 - 2.26 (1 H), 2.68 - 2.75 (2 H), 2.93 - 3.03 (2 H), 4.17 - 4.26 (2 H), 5.36 -} _{5.45 (1 H), 7.08 - 7.38 (3 H), 7.50 - 7.56 (1 H), 7.65 - 7.72 (1 H), 8.49 - 8.52 (1 H), 9.04 - 9.09 (2} H), missing proton(s) presumably hidden by/ overlapping with solvent signals. ₁₀ E_{xample E10 is prepared in analogy to example E1: Intermediate 9 (80 µmol), (S)-3-} _{methoxypyrrolidine hydrochloride 22 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane} complex (80 µmol), 2 mL MeOH. Example E10 Analytical HPLC-MS Method: A R_{[min]: 0.81 MS [m/z +} _{t ]: 531 [M+H]} Analytical SFC method: AP R_{t [min]: 1.07 d.e. > 98%} 84 12-0519-WO-1 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.49 - 1.69 (6 H), 1.89 - 2.01 (3 H), 2.17 - 2.27 (1 H),} _{2.58 - 2.66 (1 H), 2.73 - 2.80 (1 H), 2.92 - 3.03 (2 H), 3.15 - 3.19 (3 H), 3.81 - 3.89 (1 H), 4.15 -} _{4.26 (2 H), 5.36 - 5.46 (1 H), 7.07 - 7.38 (3 H), 7.49 - 7.56 (1 H), 7.65 - 7.73 (1 H), 8.47 - 8.54 (1} _{H), 9.02 - 9.10 (2 H), missing proton(s) presumably hidden by/ overlapping with solvent signals.} 9_{cis-23 cis-E11a cis-E11b} _{5 A mixture of Intermediate 9 (250 mg, 579 µmol), (1r,2s,5s)-rel-3-azabicyclo[3.1.0]hexan-2-ylmethanol} _{hydrochloride cis-23 (134 mg, 869 µmol), AcOH (68 µL, 1.16 mmol), triethylamine (117 µL, 1.16} mmol) in DMF (2.9 mL) is stirred for 4 h at rt, before sodium triacetoxyborohydride (246 mg, 1.16 mmol) is added at rt. The mixture is stirred for 2 h at rt. Water, MeOH, THF are added, the mixture filtered, and directly purified by prep. RP-HPLC (basic conditions) and chiral SFC to obtain example _{10 cis-E11a and cis-E11b as single stereoisomers. The absolute configuration of the hydroxymethyl} substituent and [3.1.0] ring system is not known; their relative configuration is cis. Example cis-E11a Analytical HPLC-MS Method: D R_{[ +} _{t min]: 0.96 MS [m/z]: 529 [M+H]} Analytical SFC method: N R_{t [min]: 4.47 d.e. > 96%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.14 - 0.25 (1 H), 0.48 - 0.56 (1 H), 1.29 - 1.37 (1 H),} _{1.47 - 1.73 (7 H), 1.78 - 1.86 (1 H), 2.69 - 2.88 (5 H), 2.98 - 3.06 (1 H), 3.18 - 3.26 (1 H), 3.54 -} _{3.62 (1 H), 4.34 - 4.48 (3 H), 5.36 - 5.46 (1 H), 7.07 - 7.38 (3 H), 7.49 - 7.57 (1 H), 7.64 - 7.73 (1} _{H), 8.46 - 8.54 (1 H), 9.02 - 9.10 (2 H).} Example cis-E11b Analytical HPLC-MS Method: D R_{[min]: 0.96 MS [m/z]: 529 [M +} _{t +H]} Analytical SFC method: N R_{t [min]: 3.53 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.15 - 0.24 (1 H), 0.48 - 0.56 (1 H), 1.28 - 1.38 (1 H),} _{1.48 - 1.72 (7 H), 1.77 - 1.86 (1 H), 2.69 - 2.89 (5 H), 2.99 - 3.07 (1 H), 3.19 - 3.27 (1 H), 3.54 -} 85 12-0519-WO-1 3_{.62 (1 H), 4.34 - 4.49 (3 H), 5.36 - 5.46 (1 H), 7.06 - 7.38 (3 H), 7.49 - 7.57 (1 H), 7.64 - 7.72 (1} _{H), 8.46 - 8.54 (1 H), 9.00 - 9.10 (2 H)} 5_{Example E12 is prepared in analogy to example E1: Intermediate 9 (80 µmol), (3S,5S)-5-} _{methylpyrrolidin-3-ol hydrochloride 24 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane} complex (80 µmol), 2 mL MeOH. Example E12 Analytical HPLC-MS Method: A R [min]: 0. + t 72 MS [m/z]: 517 [M+H] Analytical SFC method: AL R_{t [min]: 2.30 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.48 - 1.69 (6 H), 1.89 - 2.00 (3 H), 2.17 - 2.26 (1 H),} _{2.58 - 2.66 (1 H), 2.73 - 2.80 (1 H), 2.92 - 3.03 (2 H), 3.15 - 3.19 (3 H), 3.81 - 3.90 (1 H), 4.15 -} _{4.26 (2 H), 5.36 - 5.46 (1 H), 7.07 - 7.38 (3 H), 7.48 - 7.56 (1 H), 7.64 - 7.72 (1 H), 8.47 - 8.53 (1} _{H), 9.00 - 9.12 (2 H), missing proton(s) presumably hidden by/ overlapping with solvent signals.} 10 A_{mixture of Intermediate 9 (200 mg, 464 µmol), 1-[(methylamino)methyl]cyclopropan-1-ol} _{hydrochloride 25 (101 mg, 695 µmol), AcOH (54 µL, 927 µmol), triethylamine (94 mg, 927 µmol) in} DMF (2.9 mL) is stirred for 4 h at rt, before sodium triacetoxyborohydride (197 mg, 927 µmol) is added 15 at rt. The mixture is stirred for 2 h at rt. Water, MeOH, THF are added, the mixture is filtered, and directly purified by prep. RP-HPLC (basic conditions) to obtain example E13. 86 12-0519-WO-1 Example E13 Analytical HPLC-MS Method: D R [min]: 0.97 M + t S [m/z]: 517 [M+H] Analytical SFC method: T R_{t [min]: 4.34 e.e. = 90%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.35 - 0.42 (2 H), 0.52 - 0.58 (2 H), 1.48 - 1.66 (5 H),} _{1.78 - 1.89 (2 H), 2.25 - 2.35 (3 H), 2.52 - 2.55 (2 H), 2.60 - 2.70 (1 H), 2.77 - 2.86 (2 H), 4.33 -} _{4.46 (2 H), 4.78 - 4.81 (1 H), 5.37 - 5.46 (1 H), 7.07 - 7.38 (3 H), 7.49 - 7.56 (1 H), 7.66 - 7.72 (1} _{H), 8.48 - 8.55 (1 H), 9.03 - 9.10 (2 H).} 5_{A mixture of Intermediate 9 (90 mg, 209 µmol), (S)-morpholin-2-ylmethanol hydrochloride 26 (67 mg,} 417 µmol) is stirred in MeOH (2 mL) for 2 h at 35 °C. AcOH (18 µL, 313 µmol) and sodium cyanoborohydride (26 mg, 417 µmol) are added and stirred for 16 h at rt. The mixture is diluted with water and MeOH, filtered and basified with triethylamine. The mixture is purified by prep. RP-HPLC (acidic conditions) to obtain the desired compound. The compound is dissolved in MeOH und filtered 10 through a carbonate cartridge, the solvent is evaporated, dissolved in ACN/H2O and lyophilized to obtain example E14. Example E14 Analytical HPLC-MS Method: D R_{[min]: 0.73 MS [m/z]: 533 +} _{t [M+H]} Analytical SFC method: Y R_{t [min]: 5.10 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.48 - 1.64 (5 H), 1.87 - 1.97 (3 H), 2.16 - 2.25 (1 H),} _{2.34 - 2.46 (1 H), 2.70 - 2.77 (1 H), 2.81 - 2.92 (3 H), 3.35 - 3.52 (3 H), 3.74 - 3.81 (1 H), 4.31 -} _{4.43 (2 H), 4.56 - 4.63 (1 H), 5.36 - 5.46 (1 H), 7.08 - 7.38 (3 H), 7.49 - 7.56 (1 H), 7.63 - 7.73 (1} _{H), 8.48 - 8.53 (1 H), 9.03 - 9.10 (2 H), missing proton(s) presumably hidden by/ overlapping} with solvent signals. 87 12-0519-WO-1 Synthesis of example E15 E_{xample E15 is prepared in analogy to example E14: Intermediate 9 (209 µmol), (R)-morpholin-2-} _{5 ylmethanol hydrochloride 27 (417 µmol), AcOH (313 µmol), and sodium cyanoborohydride (417} µmol), 2 mL MeOH. Example E15 Analytical HPLC-MS Method: D R_{[min] +} _{t : 0.79 MS [m/z]: 533 [M+H]} Analytical SFC method: Y R_{t [min]: 4.57 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.47 - 1.65 (5 H), 1.87 - 2.00 (3 H), 2.14 - 2.26 (1 H),} _{2.34 - 2.46 (1 H), 2.69 - 2.77 (1 H), 2.82 - 2.93 (3 H), 3.35 - 3.52 (3 H), 3.72 - 3.82 (1 H), 4.31 -} _{4.44 (2 H), 4.54 - 4.64 (1 H), 5.35 - 5.47 (1 H), 7.06 - 7.38 (3 H), 7.47 - 7.57 (1 H), 7.64 - 7.72 (1} _{H), 8.47 - 8.54 (1 H), 9.00 - 9.12 (2 H), missing proton(s) presumably hidden by/ overlapping} with solvent signals. _{10 9 cis-28 cis-E16a cis-E16b} _{A mixture of intermediate 9 (100 mg, 232 µmol), cis-4-aminotetrahydrofuran-3-ol cis-28 (27 mg, 255} µmol) and AcOH (15 µL, 255 µmol) in DCM (1 mL) is stirred for 15 min at rt before sodium triacetoxyborohydride (76 mg, 348 µmol) is added. The mixture is stirred for 3 h at rt. Saturated aqueous sodium bicarbonate solution and DCM are added. The separated aqueous layer is extracted with DCM 15 and the combined organic layers are concentrated in vacuo. The material is purified by prep. RP-HPLC (_{acidic conditions) and by chiral SFC to obtain examples cis-E16a and cis-E16b as single} 88 12-0519-WO-1 stereoisomers. The absolute configuration of the amino and hydroxy substituents at the THF ring is not known; their relative configuration is cis. Example cis-E16a HPLC-MS Method: D R_{[min] +} _{t : 0.87 MS [m/z]: 519 [M+H]} Analytical SFC method: J R_{t [min]: 4.38 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.34 - 1.56 (5 H), 1.90 - 2.03 (2 H), 2.67 - 2.77 (1 H),} _{2.91 - 3.02 (2 H), 3.56 - 3.63 (1 H), 3.77 - 3.86 (2 H), 4.02 - 4.09 (1 H), 4.18 - 4.30 (2 H), 4.74 -} _{5.07 (1 H), 5.36 - 5.46 (1 H), 7.07 - 7.38 (3 H), 7.48 - 7.56 (1 H), 7.64 - 7.72 (1 H), 8.47 - 8.54 (1} _{H), 9.02 - 9.11 (2 H), missing proton(s) presumably hidden by/ overlapping with solvent signals.} Example cis-E16b HPLC-MS Method: D R_{[min]: 0.87 MS [m/ +} _{t z]: 519 [M+H]} Analytical SFC method: J R_{t [min]: 5.00 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.37 - 1.56 (5 H), 1.90 - 2.01 (2 H), 2.65 - 2.77 (1 H),} _{2.90 - 3.03 (2 H), 3.54 - 3.63 (1 H), 3.76 - 3.87 (2 H), 4.01 - 4.09 (1 H), 4.17 - 4.30 (2 H), 4.79 -} _{5.00 (1 H), 5.36 - 5.47 (1 H), 7.07 - 7.39 (3 H), 7.48 - 7.57 (1 H), 7.64 - 7.72 (1 H), 8.45 - 8.54 (1} _{H), 9.01 - 9.11 (2 H), missing proton(s) presumably hidden by/ overlapping with solvent signals.} 5 A_{mixture of Intermediate 9 (200 mg, 464 µmol), azepin-4-ol 29 (111 mg, 695 µmol), AcOH (53 µL,} 927 µmol), triethylamine (130 µL, 927 µmol) in DMF (2 mL) is stirred for 4 h at rt, before sodium 10 triacetoxyborohydride (197 mg, 927 µmol) is added at rt. The mixture is stirred for 2 h at rt. Water, MeOH, THF are added, the mixture is filtered and directly purified by prep. RP-HPLC (basic c_{onditions) and chiral SFC to obtain examples E17a andE17b as single stereoisomers. The absolute} configuration of the hydroxy substituent is not known. 89 12-0519-WO-1 Example E17a Analytical HPLC-MS Method: D R [min]: 0.93 MS [m/z]: 531 [M+ + t H] Analytical SFC method: AC R_{t [min]: 5.73 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.36 - 1.85 (13 H), 2.57 - 2.73 (4 H), 2.77 - 2.88 (2 H),} _{3.63 - 3.74 (1 H), 4.25 - 4.43 (3 H), 5.36 - 5.47 (1 H), 7.07 - 7.38 (3 H), 7.49 - 7.56 (1 H), 7.65 -} _{7.72 (1 H), 8.47 - 8.53 (1 H), 9.01 - 9.10 (2 H), missing proton(s) presumably hidden by/} overlapping with solvent signals. Example E17b Analytical HPLC-MS Method: D R_{[min +} _{t ]: 0.93 MS [m/z]: 531 [M+H]} Analytical SFC method: AC R_{t [min]: 4.48 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.37 - 1.86 (13 H), 2.58 - 2.73 (4 H), 2.77 - 2.87 (2 H),} _{3.63 - 3.74 (1 H), 4.28 - 4.44 (3 H), 5.36 - 5.46 (1 H), 7.08 - 7.40 (3 H), 7.50 - 7.57 (1 H), 7.65 -} _{7.73 (1 H), 8.50 - 8.53 (1 H), 9.03 - 9.09 (2 H), missing proton(s) presumably hidden by/} overlapping with solvent signals. 5 A_{mixture of Intermediate 9 (40 mg, 93 µmol), (S)-3-hydroxypiperidine hydrochloride 30 (20 mg, 139} _{µmol) and AcOH (5.3 µL, 93 µmol) in DMF (0.8 mL) is stirred at rt for 1 h before sodium} triacetoxyborohydride (79 mg, 371 µmol) is added. The mixture is stirred for 4 h at rt. Water is added and the mixture is basified with aqueous NH₃ solution (10 %). The mixture is further diluted with 10 THF/MeOH, filtered and directly purified by prep. RP-HPLC (basic conditions) to obtain example E18. Example E18 Analytical HPLC-MS Method: D R_{[min]: 0.93 +} _{t MS [m/z]: 517 [M+H]} 90 12-0519-WO-1 Analytical SFC method: S R_{t [min]: 7.84 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.99 - 1.15 (1 H), 1.32 - 1.45 (1 H), 1.47 - 1.69 (6 H),} _{1.75 - 1.90 (3 H), 1.90 - 2.01 (1 H), 2.01 - 2.15 (1 H), 2.69 - 2.77 (1 H), 2.78 - 2.93 (3 H), 3.38 -} _{3.51 (1 H), 4.34 - 4.47 (2 H), 4.48 - 4.59 (1 H), 5.36 - 5.47 (1 H), 7.07 - 7.39 (3 H), 7.49 - 7.58 (1} _{H), 7.65 - 7.74 (1 H), 8.49 - 8.55 (1 H), 9.00 - 9.14 (2 H), missing proton(s) presumably hidden} by/ overlapping with solvent signals. Synthesis of intermediate 33 5_{A mixture of bromide 7 (80 mg, 194 µmol), tert‐butyl 1,9‐diazaspiro[5.5]undecane‐1‐carboxylate 32} (78 mg, 290 µmol) and catalyst I (7 mg, 8 µmol) in 1,4-dioxane (3 mL) is degassed with argon. K₃PO₄ (126 mg, 581 µmol) is added, and the mixture is heated for 3.5 h at 95 °C. The mixture is filtered, rinsed w_{ith EtOAc, and concentrated in vacuo. Intermediate 33 is used directly for the next step.} Intermediate 33 Analytical HPLC-MS Method: D R_{[min]: 1.19 +} _{t MS [m/z]: 587 [M+H]} 10 Synthesis of example E19 A_{mixture of intermediate 33 (140 mg, 167 µmol) and TFA (0.4 mL) in DCM (2 mL) is stirred for 1.5} _{h at rt. The mixture is concentrated in vacuo and the residue is dissolved in ACN. Cs2CO3 is added, and} 15 the resulting suspension is filtered. The filtrate is purified by prep. RP-HPLC (basic conditions) to obtain example E19. 91 12-0519-WO-1 Example E19 Analytical HPLC-MS Method: D R_{t [min]: 1.01 MS [m/z]: 487 [M+H]}+ Chiral SFC Method: H R_{t [min]: 3.67 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.30 - 1.79 (13 H), 2.52 - 2.80 (3 H), 3.40 - 3.48 (2 H),} _{3.64 - 3.80 (2 H), 5.42 (1 H), 7.08 - 7.38 (3 H), 7.52 (1 H), 7.69 (1 H), 8.49 (1 H), 9.03 (1 H),} 9.07 (1 H). Synthesis of example E20 5_{A mixture of intermediate 7 (100 mg, 230 µmol), 1-methyl-1,8-diazaspiro[4.5]decane dihydrochloride} _{52 (60 mg, 253 µmol) and Cs2CO3 (300 mg, 920 µmol) in 1,4-dioxane (2 mL) is degassed with argon.} Then catalyst I (19 mg, 23 µmol) is added, and the mixture is heated under argon for 16 h at 120°C. Aqueous NaHCO3 solution is added, the aqueous layer is extracted with EtOAc and the combined organic layers are dried with MgSO4. After filtration and evaporation of the solvent the material is10 purified by prep. RP-HPLC (basic conditions) to obtain example E20. Example E20 Analytical HPLC-MS Method: D R_{[min]: 1.02 MS [m/z]: 487 [M+H +} _{t ]} Analytical SFC method: AD Rt [min]: 1.55 e.e. > 98% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.48 - 1.57 (3 H), 1.64 - 1.85 (6 H), 2.04 - 2.09 (1 H),} _{2.16 - 2.24 (3 H), 2.65 - 2.74 (2 H), 2.78 - 2.91 (2 H), 4.29 - 4.41 (2 H), 5.36 - 5.48 (1 H), 7.05 -} _{7.39 (3 H), 7.49 - 7.57 (1 H), 7.64 - 7.73 (1 H), 8.45 - 8.55 (1 H), 8.99 - 9.11 (2 H), missing} proton(s) presumably hidden by/ overlapping with solvent signals. 92 12-0519-WO-1 Synthesis of examples cis-E21a and cis-E21b I_{ntermediate 9 (200 mg, 464 µmol) is dissolved in THF (7.5 mL). Then rac-cis azabicyclo[3.1.0]hexan-} _{5 1-ylmethanole hydrochloride cis-34 (347 mg, 2.32 mmol), and molecular sieves 4 Å are added and} stirring is continued for 1 h at 60°C. After cooling to rt, sodium triacetoxyborohydride (202 mg, 927 µmol) is added and the reaction mixture stirred for 1 h at rt. The crude mixture is filtered through Celite and directly purified by prep. RP-HPLC (basic conditions) and by chiral SFC to obtain example cis- E_{21a and example cis-E21b as single stereoisomers. The absolute configuration of the bridged carbon} _{10 atoms is not known; the relative configuration of the [3.1.0] ring system is cis.} .Example cis-E21a Analytical HPLC-MS Method: D R_{[min] +} _{t : 0.95 MS [m/z]: 529 [M+H]} Analytical SFC method: T Rt [min]: 4.58 d.e. = 98% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 0.33 - 0.41 (1 H), 0.69 - 0.76 (1 H), 1.17 - 1.25 (1 H),} _{1.44 - 1.57 (5 H), 1.82 - 1.94 (2 H), 2.24 - 2.38 (3 H), 2.93 - 3.08 (4 H), 3.39 - 3.53 (2 H), 4.09 -} _{4.20 (2 H), 4.44 - 4.50 (1 H), 5.35 - 5.45 (1 H), 7.08 - 7.38 (3 H), 7.49 - 7.56 (1 H), 7.65 - 7.72 (1} _{H), 8.46 - 8.53 (1 H), 9.01 - 9.10 (2 H).} Example cis-E21b Analytical HPLC-MS Method: D R_{[min]: 0.95 MS [m/z]: 529 [M+ +} _{t H]} Analytical SFC method: T Rt [min]: 4.95 d.e. = 94% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 0.34 - 0.40 (1 H), 0.70 - 0.77 (1 H), 1.17 - 1.26 (1 H),} _{1.42 - 1.58 (5 H), 1.82 - 1.94 (2 H), 2.25 - 2.39 (3 H), 2.94 - 3.08 (4 H), 3.40 - 3.54 (2 H), 4.09 -} _{4.20 (2 H), 4.38 - 4.54 (1 H), 5.35 - 5.46 (1 H), 7.07 - 7.39 (3 H), 7.49 - 7.57 (1 H), 7.64 - 7.74 (1} _{H), 8.47 - 8.54 (1 H), 9.00 - 9.13 (2 H).} 93 12-0519-WO-1 Synthesis of example E22 (R)-3-Hydroxypiperidine hydrochloride 35 (130 mg, 927 µmol) is dissolved in MeOH (5 mL) and 5_{filtered through a carbonate cartridge. Intermediate 9 (200 mg, 464 µmol) is added and the mixture is} stirred for 2 h at 35 °C. AcOH (41 µl, 695 µmol) and sodium cyanoborohydride (58 mg, 927 µmol) are added and the mixture stirred for 16 h at rt. Water is added, filtered and the filtrate is directly purified b_{y prep. RP-HPLC (acidic conditions). After lyophilization, the material is redissolved in MeOH and} filtered through a carbonate cartridge to obtain example E22. Example E22 Analytical HPLC-MS Method: D R_t [min]: 0.80 MS [m/z]: 517 [M+H]+ Analytical SFC method: Z R_{t [min]: 4.85 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.00 - 1.14 (1 H), 1.30 - 1.47 (1 H), 1.49 - 1.67 (6 H),} _{1.74 - 1.89 (3 H), 1.89 - 1.98 (1 H), 2.03 - 2.13 (1 H), 2.65 - 2.77 (1 H), 2.77 - 2.92 (3 H), 3.37 -} _{3.49 (1 H), 4.32 - 4.46 (2 H), 4.49 - 4.56 (1 H), 5.36 - 5.47 (1 H), 7.07 - 7.40 (3 H), 7.48 - 7.57 (1} _{H), 7.66 - 7.73 (1 H), 8.46 - 8.53 (1 H), 9.02 - 9.09 (2 H), missing proton(s) presumably hidden} by/ overlapping with solvent signals. 10 94 Synthesis of intermediate 37

To a mixture of intermediate 5 (12.5 g, 51.6 mmol), (lR)-l-[2-fhioro-3-(trifluoromethyl)phenyl]ethan- 1-amine hydrochloride 36 (CAS: 2230840-52-3; 13.8 g, 51.6 mmol) and N-methyl morpholine (20.1 mL, 181 mmol) in ACN (152 mL) is added 1-propanephosphonic anhydride (50%; 36.9 mL, 62.0 mmol) at 0 °C dropwise. The mixture is allowed to reach rt and is stirred for 3 h. ACN is evaporated under reduced pressure. Water and EtOAc are added to the remaining material. The organic layer is separated and extracted with 0.5 M KHSO4 solution and water sequentially. The organic layer is dried with MgSO4, filtered, and the solvent is evaporated. The remaining material is dissolved in warm EtOAc and is allowed to cool down over night. The solids are filtered to obtain intermediate 37. The liquid layer is separated, and the solvent evaporated. The remaining solid is stirred in MTBE. The solids are filtered to obtain intermediate 37, and the two batches are combined.

Intermediate 37

HPLC-MS Method: E

Rt [min] : 1.04 MS [m/z] : 431 [M+H]⁺

Synthesis of intermediate 38

A mixture of intermediate 37 (1.3 g, 3.1 mmol), piperidin-4-one hydrochloride 8 (881 mg, 6.2 mmol) and CS2CO3 (3.5 g, 10.8 mmol) in 1,4-dioxane (18 mL) is degassed with argon. Catalyst I (78 mg, 93 μmol) is added, and the mixture is heated under argon for 3.5 h at 95 °C. The mixture is fdtered, the solids are washed with EtOAc, and the combined solvents are evaporated under reduced pressure. The material is purified by column chromatography (SiCh; EtOAc/cyclohexane: 80:20 a 100:0) to obtain intermediate 38. Intermediate 38

Analytical HPLC-MS Method: D

Rt [min]: 1.00 MS [m/z]: 450 [M+H]⁺ Synthesis of intermediate 39

HATU (189 mg, 496 μmol) and triethylamine (232 pL, 1.7 μmol) are added to intermediate 12 (100 mg, 413 μmol) in DMF (1.6 mL). The mixture is stirred for 15 min at rt before (lR)-l-[2-fluoro-3- (trifluoromethyl)phenyl]ethan-l -amine hydrochloride 36 (CAS: 2230840-52-3; 112 mg, 454 μmol) is added. The mixture is stirred for 16 h at rt before it is diluted with MeOH, filtered, and purified by prep. RP-HPLC (acidic conditions) to obtain intermediate 39.

Intermediate 39

HPLC-MS Method: E

Rt [min] : 0.98 MS [m/z] : 431 [M+H]⁺ 12-0519-WO-1 3_{9 37} _{A mixture of intermediate 39 (12.0 g, 19.5 mmol), Cs2CO3 (6.3 g, 19.5 mmol), piperidine (1.9 mL, 19.5} 5 mmol) in 1,4-dioxane (90 mL) is stirred at 100 °C for 16 h. EtOAc and water are added. The aqueous layer is separated and extracted with EtOAc. The combined organic layers are dried over Na2SO4. The solids are filtered, and the solvent is evaporated under reduced pressure. The material is purified by column chromatography (SiO2; EtOAc/petroleum ether: 10:90 à 30:70) to obtain intermediate 37. Intermediate 37 Analytical HPLC-MS Method: D R_t [min]: 1.00 MS [m/z]: 431 [M+H]+ 10 E_{xample E24 is prepared in analogy to example E1: Intermediate 38 (80 µmol), (3R,5S)-5-} _{methylpyrrolidin-3-ol hydrochloride 40 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane}15 complex (80 µmol), 2 mL MeOH. Example E24 HPLC-MS Method: A Rt [min]: 0.80 MS [m/z]: 535 [M+H]⁺ Analytical SFC method: L R_{t [min]: 2.25 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.42 - 1.49 (3 H), 1.50 - 1.56 (3 H), 1.62 - 1.70 (1 H),} _{1.77 - 1.92 (2 H), 2.04 - 2.12 (1 H), 2.18 - 2.27 (1 H), 2.37 - 2.45 (1 H), 2.80 - 2.90 (2 H), 3.82 -} _{3.93 (1 H), 4.36 - 4.43 (1 H), 4.43 - 4.57 (2 H), 5.38 - 5.47 (1 H), 7.14 - 7.20 (1 H), 7.36 - 7.45 (1} 97 12-0519-WO-1 H_{), 7.63 - 7.73 (1 H), 7.80 - 7.86 (1 H), 8.52 - 8.59 (1 H), 9.09 - 9.19 (2 H), 9.45 - 9.58 (1 H),} missing protons hidden by/ overlapping with by solvent signals. 5_{Example E25 is prepared in analogy to example E1: Intermediate 38 (80 µmol), (2R,3R)-2-} _{methylazetidin-3-ol hydrochloride 41 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane complex} (80 µmol), 2 mL MeOH. Example E25 HPLC-MS Method: A Rt [min]: 0.77 MS [m/z]: 521 [M+H]⁺ Analytical SFC method: Q R_{t [min]: 1.76 d.e. > 97%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.02 - 1.09 (3 H), 1.24 - 1.43 (2 H), 1.49 - 1.55 (3 H),} _{1.65 - 1.83 (2 H), 2.27 - 2.37 (1 H), 2.93 - 3.04 (3 H), 4.05 - 4.19 (3 H), 5.35 - 5.43 (1 H), 7.08 -} _{7.14 (1 H), 7.35 - 7.44 (1 H), 7.63 - 7.70 (1 H), 7.77 - 7.84 (1 H), 8.45 - 8.52 (1 H), 9.00 - 9.05 (1} H), missing proton(s) presumably hidden by/ overlapping with solvent signals. 10 E_{xample E26 is prepared in analogy to example E1: Intermediate 38 (80 µmol), (S)-pyrrolidin-3-} _{ylmethanol 42 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane complex (80 µmol), 2 mL} MeOH; purification by prep. RP-HPLC (acidic conditions). Example E26 HPLC-MS Method: A 98 12-0519-WO-1 R_t [min]: 0.78 MS [m/z]: 535 [M+H]⁺ Analytical SFC method: AP R_{t [min]: 1.67 e.e./d.e. >98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.49 - 1.88 (6 H), 1.90 - 2.24 (3 H), 2.35 - 2.46 (0.5 H),} _{2.82 - 2.93 (2.5 H), 3.07 - 3.23 (1 H), 4.42 - 4.55 (2 H), 5.36 - 5.47 (1 H), 7.15 - 7.21 (1 H), 7.38} _{- 7.45 (1 H), 7.64 - 7.72 (1 H), 7.80 - 7.88 (1 H), 8.52 - 8.58 (1 H), 9.09 - 9.20 (2 H), 9.50 - 9.76} (1 H), missing proton(s) presumably hidden by/ overlapping with solvent signals. 5_{Example E27 is prepared in analogy to example E1: Intermediate 38 (80 µmol), (R)-pyrrolidin-3-} _{ylmethanol 43 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane complex (80 µmol), 2 mL} MeOH; purification by prep. RP-HPLC (acidic conditions). Example E27 HPLC-MS Method: A Rt [min]: 0.78 MS [m/z]: 535 [M+H]⁺ Analytical SFC method: L R_{t [min]: 2.11 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.48 - 1.89 (6 H), 1.89 - 2.24 (3 H), 2.35 - 2.46 (0.5 H),} _{2.80 - 2.95 (2.5 H), 3.07 - 3.22 (1 H), 4.42 - 4.55 (2 H), 5.37 - 5.47 (1 H), 7.15 - 7.22 (1 H), 7.37} _{- 7.45 (1 H), 7.64 - 7.72 (1 H), 7.79 - 7.88 (1 H), 8.51 - 8.59 (1 H), 9.08 - 9.22 (2 H), 9.47 - 9.76} (1 H), missing proton(s) presumably hidden by/ overlapping with solvent signals. 10 3_{8 16 E28} 99 12-0519-WO-1 E_{xample E28 is prepared in analogy to example E1: Intermediate 38 (80 µmol), (7R)-5-} _{azaspiro[2.4]heptan-7-ol 16 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane complex (80} µmol), 2 mL MeOH; purification by prep. RP-HPLC (acidic conditions). Example E28 HPLC-MS Method: A R [min]: 0.80 MS + t [m/z]: 547 [M+H] Analytical SFC method: L R_{t [min]: 2.16 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.63 - 0.82 (3 H), 0.89 - 0.98 (1 H), 1.48 - 1.59 (3 H),} _{1.65 - 1.89 (2 H), 2.07 - 2.29 (2 H), 2.78 - 2.95 (2 H), 3.18 - 3.31 (1 H), 3.86 - 3.94 (1 H), 4.44 -} _{4.55 (2 H), 5.37 - 5.47 (1 H), 7.15 - 7.20 (1 H), 7.37 - 7.45 (1 H), 7.65 - 7.72 (1 H), 7.79 - 7.87 (1} _{H), 8.55 (1 H), 9.09 - 9.20 (2 H), 9.97 - 10.29 (1 H), missing proton(s) presumably hidden by/} overlapping with solvent signals. 5 Synthesis of example cis-E29 E_{xample cis-E29 is prepared in analogy to example E1: Intermediate 38 (80 µmol), diol cis-44 (0.12} _{10 mmol), AcOH (0.51 mmol), and 2-picoline-borane complex (80 µmol), 2 mL MeOH; purification by} _{prep. RP-HPLC (acidic conditions) and is obtained as a mixture of cis-diastereomers.} Example cis-E29 HPLC-MS Method: A R_{[min]: 0.76 MS [m +} _{t /z]: 565 [M+H]} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.15 - 1.21 (3 H), 1.49 - 1.58 (3 H), 1.68 - 2.03 (4 H),} _{2.04 - 2.13 (1 H), 2.16 - 2.25 (1 H), 2.81 - 2.99 (3 H), 3.01 - 3.12 (1 H), 4.44 - 4.61 (2 H), 5.12 -} _{5.31 (1 H), 5.37 - 5.47 (1 H), 7.14 - 7.20 (1 H), 7.38 - 7.45 (1 H), 7.64 - 7.73 (1 H), 7.79 - 7.87 (1} _{H), 8.53 - 8.58 (1 H), 8.74 - 8.87 (1 H), 9.08 - 9.18 (2 H), missing proton(s) presumably hidden} by/ overlapping with solvent signals. 100 12-0519-WO-1 Synthesis of example E30 3_{8 45 E30} _{Example E30 is prepared in analogy to example E1: Intermediate 38 (80 µmol), (7S)-5-} _{5 azaspiro[2.4]heptan-7-ol 45 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane complex (80} µmol), 2 mL MeOH; purification by prep. RP-HPLC (acidic conditions). Example E30 HPLC-MS Method: A R_{t [min]: 0.80 MS [m/z]: 547 [M+H]}+ Analytical SFC method: F R_{t [min]: 1.49 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.61 - 0.82 (3 H), 0.88 - 0.98 (1 H), 1.48 - 1.59 (3 H),} _{1.66 - 1.90 (2 H), 2.06 - 2.28 (2 H), 2.78 - 2.94 (2 H), 3.17 - 3.31 (1 H), 3.86 - 3.95 (1 H), 4.40 -} _{4.55 (2 H), 5.38 - 5.47 (1 H), 7.14 - 7.21 (1 H), 7.37 - 7.46 (1 H), 7.64 - 7.72 (1 H), 7.78 - 7.87 (1} _{H), 8.51 - 8.58 (1 H), 9.09 - 9.20 (2 H), 9.94 - 10.28 (1 H), missing proton(s) presumably hidden} by/ overlapping with solvent signals. ₁₀ E_{xample E31 is prepared in analogy to example E1: Intermediate 38 (80 µmol), L-prolinol 19 (0.12} mmol), AcOH (0.51 mmol), and 2-picoline-borane complex (80 µmol), 2 mL MeOH; purification by prep. RP-HPLC (acidic conditions). Example E31 HPLC-MS Method: A R₊ _{t [min]: 0.80 MS [m/z]: 535 [M+H]} 101 12-0519-WO-1 Analytical SFC method: L R_{t [min]: 2.06 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.49 - 1.57 (3 H), 1.73 - 2.00 (5 H), 2.00 - 2.10 (1 H),} _{2.10 - 2.26 (2 H), 2.82 - 2.93 (2 H), 3.23 - 3.34 (1 H), 3.38 - 3.47 (1 H), 4.48 - 4.57 (2 H), 5.38 -} _{5.47 (1 H), 7.14 - 7.20 (1 H), 7.37 - 7.45 (1 H), 7.65 - 7.72 (1 H), 7.81 - 7.87 (1 H), 8.52 - 8.57 (1} _{H), 9.10 - 9.17 (2 H), 9.17 - 9.24 (1 H), missing proton(s) presumably hidden by/ overlapping} with solvent signals. Synthesis of example E32 5 E_{xample E32 is prepared in analogy to example E1: Intermediate 38 (80 µmol), (2S,3R)-2-} _{methylazetidin-3-ol hydrochloride 46 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane complex} (80 µmol), 2 mL MeOH; purification by prep. RP-HPLC (acidic conditions). Example E32 HPLC-MS Method: A R_{[m +} _{t in]: 0.73 MS [m/z]: 521 [M+H]} Analytical SFC method: L R_{t [min]: 1.83 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.12 - 1.20 (3 H), 1.26 - 1.45 (2 H), 1.48 - 1.55 (3 H),} _{1.69 - 1.78 (1 H), 1.80 - 1.89 (1 H), 2.24 - 2.31 (1 H), 2.86 - 2.93 (1 H), 2.93 - 3.02 (2 H), 4.07 -} _{4.21 (2 H), 5.35 - 5.43 (1 H), 7.07 - 7.13 (1 H), 7.36 - 7.43 (1 H), 7.63 - 7.69 (1 H), 7.77 - 7.83 (1} _{H), 8.45 - 8.50 (1 H), 9.00 - 9.03 (1 H), missing proton(s) presumably hidden by/ overlapping} with solvent signals. 10 102 12-0519-WO-1 Synthesis of example E33 E_{xample E33 is prepared in analogy to example E1: Intermediate 38 (80 µmol), 5-azaspiro[2.4]heptane} 5 hydrochloride 21 (0.12 mmol), AcOH (0.51 mmol), and 2-picoline-borane complex (80 µmol), 2 mL MeOH; purification by prep. RP-HPLC (acidic conditions). Example E33 HPLC-MS Method: A R_{t [min]: 0.98 MS [m/z]: 531 [M+H]}+ Analytical SFC method: L R_{t [min]: 2.02 d.e. >98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.58 - 0.79 (4 H), 1.50 - 1.60 (3 H), 1.68 - 1.90 (3 H),} _{2.01 - 2.18 (2 H), 2.18 - 2.26 (1 H), 2.81 - 2.93 (2 H), 3.25 - 3.33 (2 H), 3.34 - 3.47 (2 H), 4.44 -} _{4.54 (2 H), 5.37 - 5.47 (1 H), 7.16 - 7.21 (1 H), 7.37 - 7.45 (1 H), 7.65 - 7.72 (1 H), 7.80 - 7.87 (1} _{H), 8.52 - 8.59 (1 H), 9.09 - 9.20 (2 H), 9.85 - 10.00 (1 H), missing proton(s) presumably hidden} by/ overlapping with solvent signals. ₁₀ A_{mixture of bromide 37 (200 mg, 464 µmol), tert-butyl 1,8-diazaspiro[4.5]decane-1-carboxylate} _{hydrochloride 47 (149 mg, 510 µmol) and Cs2CO3 (382 mg, 1.16 mmol) and catalyst I (19 mg, 23 µmol)} in degassed 1,4-dioxane (2 mL) is stirred under argon for 16 h at 95 °C. The mixture is diluted with 1,4- dioxane, filtered through a thiol resin, and washed with DMF/MeOH. The material is purified by prep. 15 RP-HPLC (acidic conditions) to obtain intermediate 48. 103 12-0519-WO-1 Intermediate 48 Analytical HPLC-MS Method: D R_{t [min]: 1.17 MS [m/z]: 591 [M+H]}+ 5 Intermediate 48 (134 mg, 227 µmol) in DCM (3 mL) is treated with 4 N HCl in 1,4-dioxane (142 µL) at 0 °C and stirred for 3 h at 0 °C. The solvent is evaporated under reduced pressure and the material is purified by prep. RP-HPLC (basic conditions) to obtain example E34. Example E34 Analytical HPLC-MS Method: D R_{[min +} _{t ]: 1.06 MS [m/z]: 491 [M+H]} Analytical SFC method: X R_{t [min]: 4.10 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.48 - 1.76 (11 H), 1.90 - 2.16 (1 H), 2.78 - 2.88 (2 H),} _{3.55 - 3.67 (4 H), 5.36 - 5.47 (1 H), 7.11 - 7.16 (1 H), 7.38 - 7.45 (1 H), 7.64 - 7.71 (1 H), 7.79 -} _{7.86 (1 H), 8.47 - 8.53 (1 H), 9.02 - 9.06 (1 H), 9.06 - 9.13 (1 H).} 10 A_{mixture of bromide 38 (80 mg, 186 µmol), N,N-dimethylpiperidine-4-amine 49 (29 mg, 223 µmol)} _{and Cs2CO3 (121 mg, 371 µmol) and catalyst I (16 mg, 19 µmol) in degassed 1,4-dioxane is stirred} under argon for 18 h at 105 °C. The mixture is diluted with THF and ACN/water and filtered. The15 material is purified by prep. RP-HPLC (basic conditions) to obtain example E35. 104 12-0519-WO-1 Example E35 Analytical HPLC-MS Method: A R_{t [min]: 0.83 MS [m/z]: 479 [M+H]}+ Analytical SFC method: X R_{t [min]: 4.14 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.46 - 1.61 (5 H), 1.84 - 1.93 (2 H), 2.17 - 2.22 (6 H),} _{2.23 - 2.35 (1 H), 2.80 - 2.91 (2 H), 4.30 - 4.37 (2 H), 5.36 - 5.47 (1 H), 7.14 - 7.19 (1 H), 7.36 -} _{7.44 (1 H), 7.63 - 7.70 (1 H), 7.82 - 7.90 (1 H), 8.47 - 8.52 (1 H), 9.04 - 9.39 (2 H), missing} proton(s) presumably hidden by/ overlapping with solvent signals. 5_{A mixture of bromide 37 (300 mg, 647 µmol), tert-butyl 2,8-diazaspiro[4.5]decane-2-carboxylate} _{hydrochloride 50 (246 mg, 971 µmol) and Cs2CO3 (532 mg, 1.62 mmol) and catalyst I (27 mg, 32 µmol)} in degassed 1,4-dioxane (2 mL) is stirred under argon for 18 h at 105 °C. The mixture is diluted with 1,4-dioxane, filtered through a thiol resin and washed with MeOH. The material is purified by prep. RP-HPLC (basic conditions) to obtain intermediate 51. 10 Intermediate 51 Analytical HPLC-MS Method: D R_{[min +} _{t ]: 1.15 MS [m/z]: 591 [M+H]} 105 12-0519-WO-1 I_{ntermediate 51 (370 mg, 626 µmol) in 1,4-dioxane (1 mL) is treated with 4 N HCl in 1,4-dioxane (2} mL) and stirred for 16 h at rt. The mixture is filtered through a carbonate cartridge, the solids are washed with MeOH and the combined solvents are evaporated under reduced pressure. The material is purified by prep. RP-HPLC (basic conditions) to obtain example E36. Example E36 Analytical HPLC-MS Method: D R_{t [min]: 1.06 MS [m/z]: 491 [M+H]}+ Analytical SFC method: H R_{t [min]: 4.02 e.e. = 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.49 - 1.58 (5 H), 1.60 - 1.71 (4 H), 2.59 - 2.64 (2 H),} _{2.78 - 2.85 (2 H), 3.48 - 3.63 (4 H), 5.37 - 5.47 (1 H), 7.11 - 7.20 (1 H), 7.38 - 7.44 (1 H), 7.64 -} _{7.71 (1 H), 7.80 - 7.86 (1 H), 8.48 - 8.53 (1 H), 9.04 - 9.07 (1 H), 9.07 - 9.14 (1 H missing proton(s)} presumably hidden by/ overlapping with solvent signals. 5 A_{mixture of bromide 37 (200 mg, 464 µmol), 1-methyl-1,8-diazaspiro[4.5]decane dihydrochloride 52} _{10 (122 mg, 510 µmol) and Cs2CO3 (534 mg, 1.62 mmol) and catalyst I (19 mg, 23 µmol) in degassed 1,4-} dioxane (2 mL) is stirred under argon for 16 h at 90 °C. The mixture is diluted with 1,4-dioxane, filtered through thiol resin, and washed with MeOH. The material is purified by prep. RP-HPLC (basic conditions) to obtain example E37. Example E37 Analytical HPLC-MS Method: D R_{[min]: 1.06 MS [ +} _{t m/z]: 505 [M+H]} Analytical SFC method: AB R_{t [min]: 3.63 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.28 - 1.37 (2 H), 1.49 - 1.57 (3 H), 1.65 - 1.84 (6 H),} _{2.17 - 2.23 (3 H), 2.65 - 2.74 (2 H), 2.80 - 2.91 (2 H), 4.30 - 4.40 (2 H), 5.37 - 5.48 (1 H), 7.13 -} _{7.21 (1 H), 7.37 - 7.45 (1 H), 7.64 - 7.71 (1 H), 7.79 - 7.87 (1 H), 8.47 - 8.56 (1 H), 9.04 - 9.13 (2} H). 106 Synthesis of intermediate 54

A mixture of intermediate 53 (WO2016147011; 188 mg, 465 μmol), Pd/C (5 wt.%, 50 mg) and 2 drops aqueous NHs solution (7 N) in MeOH (10 mL) is stirred under 1 bar hydrogen for 16 h at rt. The mixture is fdtered and the solids are washed with MeOH. The solvent is removed under reduced pressure and intermediate 54 is used in the next reaction step without further purification.

Intermediate 54

Analytical HPLC-MS Method: D

Rt [min] : 0.98 MS [m/z] : 271 [M+H]⁺

Synthesis of intermediate 55

A mixture of bromide 7 (80 mg, 194 μmol), intermediate 54 (60 mg, 222 μmol) and potassium phosphate (84 mg, 387 μmol) and catalyst I (8 mg, 10 μmol) in degassed 1,4-dioxane (3 mL) is stirred under argon for 18 h at 105 °C. The mixture is diluted with DCM, filtered through Celite, and washed with MeOH. The solvent is removed under reduced pressure and the material is used in the next reaction step without further purification.

Intermediate 55

Analytical HPLC-MS Method: D

Rt [min] : 1. 10 MS [m/z] : 603 [M+H]⁺ 12-0519-WO-1 Synthesis of examples E38a and E38b 5_{5 E38a E38b} _{Intermediate 55 (117 mg, 194 µmol) in 1,4-dioxane (2 mL) is treated with HCl in 1,4-dioxane (4 M;} 5 242 µL) and stirred for 6 h at rt, then aqueous HCl solution (4 M; 300 µL) is added, and the mixture stirred for 18 h before aqueous HCl solution (6 M; 500 µL) is added and the mixture is stirred for 16 h at rt. The mixture is neutralized with NaOH solution (4 M), basified with aqueous NH3 solution and extracted with DCM. The organic solvent is evaporated under reduced pressure and the material is p_{urified by prep. RP-HPLC (basic conditions) and chiral SFC to obtain examples E38a and E38b as}10 single stereoisomers. The absolute configuration at the pyrrolidine is not known. Example E38a Analytical HPLC-MS Method: D R_{[min]: 0.98 MS [m/z]: 50 +} _{t 3 [M+H]} Analytical SFC method: AI R_{t [min]: 2.02 d.e.: > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.48 - 1.74 (10 H), 2.74 - 2.88 (2 H), 2.89 - 2.95 (1 H),} _{3.13 - 3.24 (2 H), 3.54 - 3.59 (1 H), 4.01 - 4.29 (3 H), 5.35 - 5.47 (1 H), 7.06 - 7.38 (3 H), 7.48 -} _{7.56 (1 H), 7.64 - 7.74 (1 H), 8.47 - 8.53 (1 H), 9.01 - 9.11 (2 H), missing proton(s) presumably} hidden by/ overlapping with solvent signals. Example E38b Analytical HPLC-MS Method: D R_{[min]: 0.98 MS [m/z]: 503 [M+H]+} _t Analytical SFC method: AI R_{t [min]: 1.38 d.e. = 96%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.48 - 1.74 (11 H), 2.72 - 2.86 (2 H), 2.86 - 2.92 (1 H),} _{3.12 - 3.22 (2 H), 3.83 - 4.34 (3 H), 5.36 - 5.47 (1 H), 7.06 - 7.38 (3 H), 7.49 - 7.57 (1 H), 7.64 -} _{7.74 (1 H), 8.46 - 8.53 (1 H), 9.01 - 9.10 (2 H), missing proton(s) presumably hidden by/} overlapping with solvent signals. 108 12-0519-WO-1 A_{mixture of intermediate 38 (50 mg, 111 µmol), (2R)-1-aminopropan-2-ol 56 (13 mg, 167 µmol) and} _{5 AcOH (9.6 µL, 167 µmol) in DCM (0.5 mL) is stirred for 15 min at rt before sodium} triacetoxyborohydride (36 mg, 167 µmmol) is added. The mixture is stirred for 3 h at rt, diluted with DCM, and extracted with aqueous saturated NaHCO3 solution. The volume of the separated organic layer is decreased under reduced pressure. The material is purified by prep. RP-HPLC (acidic conditions). After lyophilization, the material is redissolved in MeOH and filtered through a carbonate_{10 cartridge to obtain example E39.} Example E39 Analytical HPLC-MS Method: D R_{[min] +} _{t : 0.97 MS [m/z]: 509 [M+H]} Analytical SFC method: AO R_{t [min]: 5.90 d.e. > 98 %} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.02 - 1.09 (3 H), 1.36 - 1.48 (2 H), 1.51 - 1.57 (4 H),} _{1.89 - 2.00 (2 H), 2.41 - 2.49 (1 H), 2.58 - 2.70 (1 H), 2.92 - 3.04 (2 H), 3.60 - 3.70 (1 H), 4.16 -} _{4.29 (2 H), 4.37 - 4.57 (1 H), 5.36 - 5.47 (1 H), 7.10 - 7.18 (1 H), 7.38 - 7.46 (1 H), 7.64 - 7.72 (1} _{H), 7.80 - 7.87 (1 H), 8.49 - 8.54 (1 H), 9.05 - 9.16 (2 H), missing proton(s) presumably hidden} by/ overlapping with solvent signals. _{15 Example E40 is prepared in analogy to example E39: Intermediate 38 (111 µmol), (2S)-1-aminopropan-} _{2-ol 57 (167 µmol), AcOH (167 µmol), and sodium triacetoxyborohydride (167 µmol), 0.5 mL DCM.} 109 12-0519-WO-1 Example E40 Analytical HPLC-MS Method: D R [min]: 0.97 MS [m + t /z]: 509 [M+H] Analytical SFC method: AG R_{t [min]: 5.00 d.e. > 98 %} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.00 - 1.09 (3 H), 1.31 - 1.48 (2 H), 1.48 - 1.67 (4 H),} _{1.88 - 1.99 (2 H), 2.42 - 2.48 (1 H), 2.56 - 2.65 (1 H), 2.93 - 3.04 (2 H), 3.58 - 3.69 (1 H), 4.14 -} _{4.27 (2 H), 4.39 - 4.44 (1 H), 5.36 - 5.47 (1 H), 7.10 - 7.16 (1 H), 7.37 - 7.45 (1 H), 7.64 - 7.71 (1} _{H), 7.79 - 7.87 (1 H), 8.48 - 8.53 (1 H), 9.04 - 9.13 (2 H), missing proton(s) presumably hidden} by/ overlapping with solvent signals. Synthesis of examples cis-E41a and cis-E41b 5_{Examples cis-E41 and cis-E41b are prepared in analogy to example E39: Intermediate 38 (223 µmol),} _{cis-4-aminotetrahydrofuran-3-ol cis-28 (245 µmol), AcOH (245 µmol), and sodium} triacetoxyborohydride (334 µmol), 1 mL DCM; in addition: chiral SFC separation (basic conditions). C_{is-E41a and cis-E41b are isolated as single stereoisomers. The absolute configuration of the amino} and hydroxy substituents at the THF ring is not known; their relative configuration is cis. Example cis-E41a Analytical HPLC-MS Method: D Rt [min]: 0.83 MS [m/z]: 537 [M+H]⁺ Analytical SFC method: J R_{t [min]: 3.22 d.e. = 97 %} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.36 - 1.58 (5 H), 1.89 - 2.03 (2 H), 2.67 - 2.77 (1 H),} _{2.92 - 3.03 (2 H), 3.57 - 3.64 (1 H), 3.76 - 3.87 (2 H), 4.03 - 4.09 (1 H), 4.20 - 4.30 (2 H), 4.68 -} _{5.11 (1 H), 5.37 - 5.48 (1 H), 7.12 - 7.17 (1 H), 7.38 - 7.45 (1 H), 7.64 - 7.72 (1 H), 7.80 - 7.87 (1} _{H), 8.48 - 8.55 (1 H), 9.04 - 9.16 (2 H), missing proton(s) presumably hidden by/ overlapping} with solvent signals. 10 Example cis-E41b Analytical HPLC-MS Method: D 110 12-0519-WO-1 R_{[min]: 0. +} _{t 83 MS [m/z]: 537 [M+H]} Analytical SFC method: J R_{t [min]: 3.61 d.e. = 84 %} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.38 - 1.58 (5 H), 1.90 - 2.03 (2 H), 2.69 - 2.79 (1 H),} _{2.92 - 3.03 (2 H), 3.56 - 3.63 (1 H), 3.76 - 3.87 (2 H), 4.04 - 4.10 (1 H), 4.20 - 4.31 (2 H), 5.37 -} _{5.47 (1 H), 7.12 - 7.16 (1 H), 7.39 - 7.45 (1 H), 7.64 - 7.72 (1 H), 7.80 - 7.87 (1 H), 8.47 - 8.54 (1} _{H), 9.05 - 9.15 (2 H), missing proton(s) presumably hidden by/ overlapping with solvent signals.} 3_{8 trans-28 trans-E42a trans-E42b} _{5 Examples trans-E42 and trans-E42b are prepared in analogy to example E39: Intermediate 38 (445} _{µmol), trans-4-aminotetrahydrofuran-3-ol trans-28 (668 µmol), AcOH (668 µmol), and sodium} triacetoxyborohydride (668 µmol), 2 mL DCM; in addition: chiral SFC separation (basic conditions). T_{rans-E42a and trans-E42b are isolated as single stereoisomers. The absolute configuration of the} amino and hydroxy substituents at the THF ring is not known; their relative configuration is trans. Example trans-E42a HPLC-MS Method: E R_t [min]: 0.83 MS [m/z]: 537 [M+H]⁺ Analytical SFC method: G R_{t [min]: 3.64 d.e. > 98 %} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.33 - 1.48 (2 H), 1.49 - 1.57 (3 H), 1.91 - 2.03 (2 H),} _{2.69 - 2.79 (1 H), 2.90 - 3.01 (2 H), 3.15 - 3.22 (1 H), 3.37 - 3.43 (1 H), 3.43 - 3.50 (1 H), 3.77 -} _{3.84 (1 H), 3.86 - 3.92 (1 H), 3.94 - 3.99 (1 H), 4.20 - 4.30 (2 H), 4.87 - 4.96 (1 H), 5.37 - 5.47 (1} _{H), 7.10 - 7.15 (1 H), 7.37 - 7.44 (1 H), 7.64 - 7.71 (1 H), 7.79 - 7.87 (1 H), 8.49 - 8.53 (1 H),} _{9.03 - 9.14 (2 H), missing proton(s) presumably hidden by/ overlapping with solvent signals.} 10 Example trans-E42b Analytical HPLC-MS Method: C Rt [min]: 0.83 MS [m/z]: 537 [M+H]⁺ Analytical SFC method: G R_{t [min]: 5.52 d.e. > 98 %} 111 12-0519-WO-1 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.33 - 1.48 (2 H), 1.49 - 1.58 (3 H), 1.92 - 2.03 (2 H),} _{2.68 - 2.79 (1 H), 2.90 - 3.02 (2 H), 3.15 - 3.22 (1 H), 3.37 - 3.44 (1 H), 3.44 - 3.50 (1 H), 3.77 -} _{3.84 (1 H), 3.86 - 3.93 (1 H), 3.93 - 4.01 (1 H), 4.20 - 4.31 (2 H), 4.85 - 4.96 (1 H), 5.36 - 5.47 (1} _{H), 7.10 - 7.17 (1 H), 7.38 - 7.45 (1 H), 7.64 - 7.71 (1 H), 7.79 - 7.86 (1 H), 8.48 - 8.54 (1 H),} _{9.03 - 9.14 (2 H), missing proton(s) presumably hidden by/ overlapping with solvent signals.} 3_{8 29 E43a E43b} _{5 Examples E43a and E43b are prepared in analogy to example E39: Intermediate 38 (556 µmol), azepin-} _{4-ol 29 (834 µmol), AcOH (834 µmol), and sodium triacetoxyborohydride (1.11 mmol), 2 mL DCM;} _{in addition: chiral SFC separation (basic conditions). E43a and E43b are isolated as single} stereoisomers. The absolute configuration of the hydroxy substituent is not known. Example E43a HPLC-MS Method: E R_{t [min]: 0.83 MS [m/z]: 549 [M+H]}+ Analytical SFC method: AC R_{t [min]: 4.46 d.e. > 98 %} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.36 - 1.85 (13 H), 2.58 - 2.73 (4 H), 2.77 - 2.88 (2 H),} _{3.63 - 3.74 (1 H), 4.17 - 4.47 (3 H), 5.35 - 5.46 (1 H), 7.10 - 7.15 (1 H), 7.38 - 7.44 (1 H), 7.64 -} _{7.71 (1 H), 7.79 - 7.87 (1 H), 8.48 - 8.52 (1 H), 9.03 - 9.14 (2 H), missing proton(s) presumably} hidden by/ overlapping with solvent signals. Example E43b HPLC-MS Method: E R_t [min]: 0.83 MS [m/z]: 549 [M+H]⁺ Analytical SFC method: AC R_{t [min]: 3.51 d.e. > 98 %} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.35 - 1.90 (13 H), 2.56 - 2.75 (4 H), 2.75 - 2.89 (2 H),} _{3.61 - 3.76 (1 H), 4.16 - 4.47 (3 H), 5.37 - 5.48 (1 H), 7.08 - 7.16 (1 H), 7.36 - 7.45 (1 H), 7.60 -} _{7.73 (1 H), 7.78 - 7.88 (1 H), 8.44 - 8.59 (1 H), 9.00 - 9.18 (2 H), missing proton(s) presumably} hidden by/ overlapping with solvent signals. 112 12-0519-WO-1 Synthesis of example E44 2_{-Picoline-borane complex (6.2 mg, 60 µmol) is added to a mixture of intermediate 38 (26 mg, 158} _{5 µmol), (S)-3-hydroxypiperidine hydrochloride 30 (17 mg, 116 µmol) and AcOH (10 µL, 514 µM) in} MeOH (0.5 mL). The mixture is stirred for 16 h at rt. The mixture is neutralized with AcOH, diluted with MeOH and purified by prep. RP-HPLC (acidic conditions) to obtain example E44. Example E44 Analytical HPLC-MS Method: E R_{[min]: 0. +} _{t 79 MS [m/z]: 535 [M+H]} Analytical SFC method: AE R_{t [min]: 1.48 d.e. > 98 %} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.50 - 2.25 (11 H), 2.80 - 2.94 (3 H), 2.98 - 3.14 (2 H),} _{4.53 (2 H), 5.42 (1 H), 7.18 (1 H), 7.41 (1 H), 7.64 - 7.72 (1 H), 7.83 (1 H), 8.55 (1 H), 8.90 -} 9.34 (3 H), missing proton(s) presumably hidden by/ overlapping with solvent signals. 10 A mixture of bromide 37 (120 mg, 278 µmol), tert‐butyl 1,9‐diazaspiro[5.5]undecane‐1‐carboxylate 32 (112 mg, 418 µmol) and Cs₂CO₃ (272 mg, 835 µmol) in 1,4-dioxane (4 mL) is degassed with argon. Catalyst I (14 mg, 17 µmol) is added, and the mixture is heated under argon for 3 h at 90 °C. The 15 mixture is diluted with ACN, filtered, and the solvent is evaporated under reduced pressure. The desired material is used directly for the next step without further purification. Intermediate 58 Analytical HPLC-MS Method: D R_{[min]: 1.23 MS +} _{t [m/z]: 605 [M+H]} 113 12-0519-WO-1 5_{8 E45} _{A mixture of intermediate 58 (168 mg, 278 µmol) and TFA (0.4 mL) in DCM (1.5 mL) is stirred for} 5 1.5 h at rt. The mixture is concentrated under reduced pressure, saturated aqueous Na2CO3 solution is added, and the mixture is extracted with EtOAc. The combined organic layers are washed with saturated NaCl solution, dried over MgSO4, and concentrated under reduced pressure. The residue is purified by prep. RP-HPLC (basic conditions) to give example E45. Example E45 HPLC-MS Method: E R_{[min]: 0.89 MS [ +} _{t m/z]: 505 [M+H]} Analytical SFC method: AL R_{t [min]: 2.75 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.38 (4 H), 1.51 - 1.78 (10 H), 2.64 - 2.70 (2 H), 3.40 -} _{3.48 (2 H), 3.70 - 3.81 (2 H), 5.39 - 5.46 (1 H), 7.12 - 7.16 (1 H), 7.39 - 7.43 (1 H), 7.66 - 7.69 (1} _{H), 7.81 - 7.84 (1 H), 8.49 (1 H), 9.02 - 9.04 (1 H), 9.08 - 9.10 (1 H).} 10 E_{xample E46 is prepared in analogy to example E39: E45 (142 µmol), aqueous formaldehyde solution} _{(37 %, 213 µmol), AcOH (142 µmol), and sodium triacetoxyborohydride (213 µmol), 3 mL DCE;}15 purification by prep. RP-HPLC (acidic conditions). Example E46 Analytical HPLC-MS Method: D R₊ _{t [min]: 1.09 MS [m/z]: 519 [M+H]} 114 12-0519-WO-1 Analytical SFC method: AL R_t [min]: 2.57 e.e. > 98% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.53 - 1.55 (3 H), 1.58 - 1.71 (3 H), 1.74 - 1.97 (3 H),} _{1.99 - 2.21 (4 H), 2.80 - 2.82 (3 H), 2.95 - 3.24 (3 H), 3.32 - 3.48 (1 H), 5.39 - 5.46 (1 H), 7.17 -} _{7.20 (1 H), 7.39 - 7.43 (1 H), 7.66 - 7.70 (1 H), 7.82 - 7.85 (1 H), 8.55 (1 H), 9.13 - 9.15 (2 H),} 9.41 (1 H), missing proton(s) presumably hidden by/ overlapping with solvent signals. 5_{Examples E47a and E47b are prepared in analogy to example E1: Intermediate 38 (223 µmol), 3-} methylpiperidin-3-ol 59 (334 µmol), AcOH (445 µmol), and 2-picoline-borane complex (445 µmol), 2 m_{L MeOH; reaction time: 16 h and in addition: purification by chiral SFC (basic conditions). E47a and} _{E47b are isolated as single stereoisomers. The absolute configuration of the hydroxy substituent is not} known. Example E47a Analytical HPLC-MS Method: D Rt [min]: 1.08 MS [m/z]: 549 [M+H]⁺ Analytical SFC method: P Rt [min]: 4.80 d.e. > 96% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.08 - 1.12 (3 H), 1.28 - 1.48 (3 H), 1.50 - 1.68 (6 H), 1.78 -} _{1.88 (2 H), 2.21 - 2.34 (2 H), 2.37 - 2.46 (2 H), 2.79 - 2.91 (2 H), 3.97 - 4.06 (1 H), 4.31 - 4.44 (2 H),} _{5.36 - 5.48 (1 H), 7.10 - 7.15 (1 H), 7.37 - 7.45 (1 H), 7.64 - 7.71 (1 H), 7.80 - 7.86 (1 H), 8.48 - 8.54 (1} _{H), 9.05 - 9.14 (2 H), missing proton(s) presumably hidden by/ overlapping with solvent signals.} 10 Example E47b Analytical HPLC-MS Method: D R_t [min]: 1.08 MS [m/z]: 549 [M+H]⁺ Analytical SFC method: P R_t [min]: 5.40 d.e. > 93% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.08 - 1.12 (3 H), 1.29 - 1.49 (3 H), 1.50 - 1.66 (6 H), 1.76 -} _{1.88 (2 H), 2.21 - 2.35 (2 H), 2.37 - 2.46 (2 H), 2.79 - 2.90 (2 H), 3.98 - 4.05 (1 H), 4.31 - 4.44 (2 H),} 115 12-0519-WO-1 5_{.37 - 5.47 (1 H), 7.10 - 7.16 (1 H), 7.37 - 7.45 (1 H), 7.64 - 7.71 (1 H), 7.79 - 7.87 (1 H), 8.48 - 8.53 (1} _{H), 9.05 - 9.13 (2 H), missing proton(s) presumably hidden by/ overlapping with solvent signals.} Synthesis of example E48 5 A_{mixture of intermediate 37 (50 mg, 116 µmol), 4-(1-pyrrolidinyl)piperidine 60 (36 mg, 232 µmol),} Cs2CO3 (94 mg, 290 µmol) in 1,4-dioxane (4 mL) is degassed with argon. Catalyst I (4 mg, 5 µmol) is added, and the mixture is stirred under argon at 90 °C for 16 h. The mixture is filtered and purified by prep. RP-HPLC (basic conditions) to obtain example E48. Example E48 HPLC-MS Method: E R_t [min]: 0.90 MS [m/z]: 505 [M+H]⁺ Analytical SFC method: S R_t [min]: 7.15 e.e. > 98% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.49 - 1.76 (9 H), 1.92 - 2.02 (2 H), 2.15 - 2.25 (1 H), 2.93 -} _{3.04 (2 H), 4.14 - 4.27 (2 H), 5.35 - 5.47 (1 H), 7.11 - 7.17 (1 H), 7.37 - 7.45 (1 H), 7.63 - 7.71 (1 H),} _{7.79 - 7.87 (1 H), 8.48 - 8.52 (1 H), 9.04 - 9.08 (1 H), 9.08 - 9.16 (1 H), missing proton(s) presumably} hidden by/ overlapping with solvent signals. 10 Synthesis of example E49 E_{xample E49 is prepared in analogy to example E48: Intermediate 37 (116 µmol), 4-}15 piperidinopiperidine 61 (232 µmol), Cs2CO3 (290 µmol), and catalyst I (5 µmol), 2 mL 1,4-dioxane. 116 12-0519-WO-1 Example E49 HPLC-MS Method: E R [min]: 0.92 + t MS [m/z]: 519 [M+H] Analytical SFC method: S R_{t [min]: 7.55 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.32 - 1.71 (11 H), 1.81 - 1.89 (2 H), 2.38 - 2.49 (4 H),} _{2.75 - 2.87 (2 H), 4.33 - 4.45 (2 H), 5.37 - 5.47 (1 H), 7.10 - 7.14 (1 H), 7.37 - 7.44 (1 H), 7.64 -} _{7.71 (1 H), 7.79 - 7.87 (1 H), 8.47 - 8.55 (1 H), 9.04 - 9.08 (1 H), 9.08 - 9.16 (1 H), missing} proton(s) presumably hidden by/ overlapping with solvent signals. 5_{2-Picoline borane complex (12 mg, 111 µmol) is added to a mixture of intermediate 38 (50 mg, 111} _{µmol), 1-amino-2-methyl-propan-2-ol 62 (21 mg, 223 µmol) and AcOH (60 µL, 1.03 mmol) in MeOH} (1 mL) and the resulting mixture is stirred for 16 h at rt. The mixture is neutralized with aqueous NaOH solution (4 N), diluted with MeOH, and purified by prep. RP-HPLC (basic conditions) to obtain example E50. Example E50 HPLC-MS Method: E R_{[min]: 0.89 MS [m/z]: 523 [M +} _{t +H]} Analytical SFC method: V R_{t [min]: 6.91 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.02 - 1.15 (6 H), 1.35 - 1.57 (6 H), 1.89 - 2.00 (2 H),} _{2.42 - 2.48 (2 H), 2.55 - 2.64 (1 H), 2.94 - 3.05 (2 H), 4.09 - 4.17 (1 H), 4.17 - 4.29 (2 H), 5.37 -} _{5.48 (1 H), 7.07 - 7.17 (1 H), 7.38 - 7.44 (1 H), 7.64 - 7.72 (1 H), 7.79 - 7.87 (1 H), 8.49 - 8.57 (1} _{H), 9.03 - 9.08 (1 H), 9.08 - 9.16 (1 H).} 10 117 12-0519-WO-1 Synthesis of intermediate cis-64 A_{mixture of intermediate 37 (130 mg, 271 µmol), tert-butyl octahydro-1H-pyrrolo[2,3-c]pyridine-1-} _{5 carboxylate cis-63 (84 mg, 353 µmol) is degassed with argon. Cs2CO3 (354 mg, 1.1 mmol) in 1,4-} dioxane (2.3 mL) and catalyst I (22.8 mg, 27 µmol) are added, and the mixture is heated under argon for 16 h at 120°C. The mixture is diluted with MeOH, filtered, and purified by prep. RP-HPLC (basic c_{onditions) to give intermediate cis-64 as mixture of stereoisomers.} Intermediate cis-64 Analytical HPLC-MS Method: D R₊ _{t [min]: 1.15 MS [m/z]: 577 [M+H]} 10 A_{mixture of intermediate cis-64 (100 mg, 173 µmol) in 1,4-dioxane (0.3 mL) is treated with HCl in} 1,4-dioxane (4 M; 434 µL) and stirring is continued for 2 h at rt. The mixture is diluted with MeOH, 15 neutralized with aqueous NH3 solution, and purified by prep. RP-HPLC (basic conditions) and chiral S_{FC to obtain example cis-E52a and example cis-E52b as single stereoisomers. The absolute} configuration of the bridgehead carbon atoms is not known; their relative configuration is cis. Example cis-E52a Analytical HPLC-MS Method: D R_{[min]: 1.00 MS [m +} _{t /z]: 477 [M+H]} Analytical SFC method: AJ R_t [min]: 2.93 d.e. > 97% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.40 - 1.57 (4 H), 1.68 - 1.94 (3 H), 2.19 - 2.30 (1 H),} _{2.75 - 2.84 (1 H), 2.91 - 3.02 (1 H), 3.10 - 3.16 (1 H), 3.38 - 3.47 (1 H), 3.67 - 3.77 (1 H), 3.93 -} 118 12-0519-WO-1 4_{.03 (1 H), 5.36 - 5.47 (1 H), 7.05 - 7.09 (1 H), 7.37 - 7.45 (1 H), 7.64 - 7.72 (1 H), 7.79 - 7.86 (1} _{H), 8.47 - 8.51 (1 H), 8.98 - 9.03 (1 H), 9.04 - 9.12 (1 H), missing proton(s) presumably hidden} by/ overlapping with solvent signals. Example cis-E52b Analytical HPLC-MS Method: D R_{t [min]: 1.00 MS [m/z]: 477 [M+H]}+ Analytical SFC method: AJ R_t [min]: 3.30 d.e. > 89% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.38 - 1.58 (4 H), 1.66 - 1.95 (3 H), 2.19 - 2.30 (1 H),} _{2.73 - 2.84 (1 H), 2.91 - 3.00 (1 H), 3.09 - 3.15 (1 H), 3.37 - 3.47 (1 H), 3.70 - 3.79 (1 H), 3.93 -} _{4.02 (1 H), 5.34 - 5.47 (1 H), 7.05 - 7.08 (1 H), 7.37 - 7.46 (1 H), 7.63 - 7.71 (1 H), 7.77 - 7.87 (1} _{H), 8.47 - 8.51 (1 H), 8.98 - 9.01 (1 H), 9.05 - 9.10 (1 H), missing proton(s) presumably hidden} by/ overlapping with solvent signals. 5_{38 trans-65 trans-E53a trans-E53b} _{Intermediate 38 (200 mg, 445 µmol) is dissolved in THF (7.2 mL) and 2-amino-1-methylcyclopentan-} 1-ol trans-65 (103 mg, 890 µmol), AcOH (51 µL, 890 µmol) and molecular sieves 4 Å are added and the mixture is stirred for 1 h at 60 °C. After cooling to rt, sodium triacetoxyborohydride (194 mg, 890 µmol) is added and the reaction mixture is stirred 1 h at rt. The reaction mixture is filtered through 10 Celite, the solvent is evaporated, and the mixture is purified by prep. RP-HPLC (basic conditions) and b_{y chiral SFC to obtain example trans-E53a and example trans-E53b as single stereoisomers. The} absolute configuration of the amino and hydroxy substituents is not known; their relative configuration is trans. Example trans-E53a Analytical HPLC-MS Method: D R_{[min]: +} _{t 1.01 MS [m/z]: 549 [M+H]} Analytical SFC method: AE R_{t [min]: 0.85 d.e. > 98%} 119 12-0519-WO-1 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.03 - 1.13 (3 H), 1.17 - 1.64 (11 H), 1.86 - 2.02 (3 H),} _{2.68 - 2.79 (1 H), 2.84 - 2.92 (1 H), 2.92 - 3.02 (2 H), 4.13 - 4.32 (3 H), 5.35 - 5.47 (1 H), 7.10 -} _{7.16 (1 H), 7.37 - 7.44 (1 H), 7.64 - 7.72 (1 H), 7.79 - 7.88 (1 H), 8.46 - 8.55 (1 H), 9.03 - 9.07 (1} _{H), 9.07 - 9.15 (1 H).} Example trans-E53b Analytical HPLC-MS Method: D R_{t [min]: 1.01 MS [m/z]: 549 [M+H]}+ Analytical SFC method: AE R_{t [min]: 1.17 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.02 - 1.11 (3 H), 1.17 - 1.61 (11 H), 1.83 - 2.02 (3 H),} _{2.68 - 2.79 (1 H), 2.84 - 2.91 (1 H), 2.91 - 3.02 (2 H), 4.17 - 4.29 (3 H), 5.37 - 5.47 (1 H), 7.08 -} _{7.17 (1 H), 7.37 - 7.45 (1 H), 7.64 - 7.72 (1 H), 7.79 - 7.87 (1 H), 8.48 - 8.53 (1 H), 9.03 - 9.07 (1} _{H), 9.07 - 9.13 (1 H), missing proton(s) presumably hidden by/ overlapping with solvent signals.} ₅ E_{xample E54 is prepared in analogy to example E1: Intermediate 38 (111 µmol), D-alaninol 66 (223} µmol), AcOH (1.03 mmol), and 2-picoline-borane complex (200 µmol), 2 mL MeOH; purification prep. RP-HPLC (basic conditions). Example E54 Analytical HPLC-MS Method: D Rt [min]: 0.96 MS [m/z]: 509 [M+H]⁺ Analytical SFC method: AH R_{t [min]: 5.51 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.90 - 0.98 (3 H), 1.28 - 1.48 (2 H), 1.50 - 1.57 (3 H),} _{1.87 - 2.01 (2 H), 2.71 - 2.85 (2 H), 2.92 - 3.03 (2 H), 3.22 - 3.26 (2 H), 4.16 - 4.30 (2 H), 4.33 -} _{4.57 (1 H), 5.37 - 5.48 (1 H), 7.11 - 7.17 (1 H), 7.39 - 7.45 (1 H), 7.64 - 7.73 (1 H), 7.80 - 7.87 (1} _{H), 8.48 - 8.55 (1 H), 9.03 - 9.08 (1 H), 9.09 - 9.13 (1 H), missing proton(s) presumably hidden} by/ overlapping with solvent signals. 120 12-0519-WO-1 E_{xample E55 is prepared in analogy to example E1: Intermediate 38 (49 µmol), 2-} _{5 (methylamino)ethanol 67 (98 µmol), AcOH (514 µmol), and 2-picoline-borane complex (49 µmol), 0.5} mL MeOH; reaction time: 16 h; purification by prep. RP-HPLC (acidic conditions). Example E55 HPLC-MS Method: E R_t [min]: 0.79 MS [m/z]: 509 [M+H]+ Analytical SFC method: X R_{t [min]: 4.62 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.46 - 1.59 (3 H), 1.76 - 1.90 (2 H), 2.08 - 2.21 (2 H),} _{2.76 - 2.82 (3 H), 2.83 - 2.95 (2 H), 3.04 - 3.13 (1 H), 3.32 - 3.37 (1 H), 4.48 - 4.59 (2 H), 5.38 -} _{5.47 (1 H), 7.15 - 7.20 (1 H), 7.37 - 7.46 (1 H), 7.64 - 7.71 (1 H), 7.79 - 7.87 (1 H), 8.50 - 8.58 (1} _{H), 9.09 - 9.17 (2 H), 9.20 - 9.29 (1 H), missing proton(s) presumably hidden by/ overlapping} with solvent signals. ₁₀ E_{xample E57 is prepared in analogy to example E1: Intermediate 38 (89 µmol), (R)-3-} _{hydroxypyrrolidine 11 (178 µmol), AcOH (1.03 µmol), and 2-picoline-borane complex (89 µmol), 0.6} mL MeOH; purification by prep. RP-HPLC (basic conditions). Example E57 HPLC-MS Method: E R_t [min]: 0.79 MS [m/z]: 521 [M+H]⁺ Analytical SFC method: AN 121 12-0519-WO-1 R_{t [min]: 1.14 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.47 - 1.60 (6 H), 1.89 - 2.01 (3 H), 2.17 - 2.26 (1 H),} _{2.35 - 2.41 (1 H), 2.60 - 2.69 (1 H), 2.74 - 2.82 (1 H), 2.93 - 3.03 (2 H), 4.13 - 4.27 (3 H), 4.43 -} _{4.86 (1 H), 5.36 - 5.48 (1 H), 7.10 - 7.18 (1 H), 7.38 - 7.45 (1 H), 7.63 - 7.71 (1 H), 7.79 - 7.87 (1} _{H), 8.47 - 8.54 (1 H), 9.04 - 9.08 (1 H), 9.08 - 9.16 (1 H), missing proton(s) presumably hidden} by/ overlapping with solvent signals. Synthesis scheme of intermediate 71 5 Synthesis of intermediate 69 A_{n aqueous Na2CO3 solution (2 M; 7.7 mL, 15 mmol), intermediate 7 (2.0 g, 4.8 mmol) and borane 68} (1.6 g, 5.8 mmol) in 1,4-dioxane (24 mL) is stirred under argon atmosphere. Catalyst II (198 mg, 242 10 µmol) is added, and the mixture is stirred at 95 °C for 1.5 h. Aqueous NaCl solution is added. The layers are separated, and the aqueous layer is extracted with EtOAc. The combined organic layers are washed with saturated NaCl solution, dried over Na2SO4, filtered and the solvent is evaporated. The residue is 122 purified by column chromatography (SiCL; EtOAc/cyclohexane: 60:40 a 100:0) to obtain intermediate

69.

Intermediate 69

Analytical HPLC-MS Method: D

Rt [min]: 0.99 MS [m/z]: 473 [M+H]⁺

Synthesis of intermediate 70

A mixture of intermediate 69 (1.6 g, 3.4 mmol) and Pd/C (I0%w/w, 200 mg) in MeOH (30 mL) is stirred under H2 atmosphere (50 psi) for 18 h at rt. The mixture is filtered, and the solvent evaporated to obtain intermediate 70. The material is used in the next reaction step without further purification.

Intermediate 70

HPLC-MS Method: E

Rt [min] : 1.02 MS [m/z] : 475 [M+H]⁺

Intermediate 70 (1.5 g, 3.2 mmol) in THF (20 mL) is treated with aqueous HC1 solution (4 M; 5 mb, 20 mmol). The mixture is stirred for 24 h at rt. An aqueous NaOH solution (4 M; 5 mL) and saturated NaHCOs solution are added, and the mixture is extracted with EtOAc. The separated organic layers are extracted with aqueous saturated NaCl solution. The solvents are evaporated under reduced pressure. The desired material is treated with MTBE and the solvent evaporated to obtain intermediate 71. The material is used in the next reaction step without further purification. 12-0519-WO-1 Intermediate 71 Analytical HPLC-MS Method: D R [min]: 0.93 + t MS [m/z]: 431 [M+H] 5_{A mixture of intermediate 71 (160 mg, 372 µmol) and (R)-3-hydroxypiperidine hydrochloride 35 (104} mg, 743 µmol) in MeOH (3 mL) is prepared and AcOH (150 µL, 2.6 mmol) added followed by 2- picoline-borane complex (49 mg, 446 µmol). The mixture is stirred for 1.5 h at rt. An aqueous NaOH solution (4 M) and MeOH are added. The mixture is filtered and directly purified by prep. RP-HPLC (basic conditions) to obtain example E58. Example E58 Analytical HPLC-MS Method: D R_t [min]: 0.95 MS [m/z]: 516 [M+H]+ Chiral SFC Method: Y R_{t [min]: 5.21 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.99 - 1.11 (1 H), 1.32 - 1.51 (3 H), 1.51 - 1.56 (3 H),} _{1.56 - 1.84 (4 H), 1.84 - 1.93 (2 H), 1.94 - 2.06 (3 H), 2.08 - 2.18 (1 H), 2.38 - 2.48 (1 H), 2.65 -} _{2.73 (1 H), 2.83 - 2.92 (1 H), 3.02 - 3.14 (1 H), 3.38 - 3.48 (1 H), 4.47 - 4.55 (1 H), 5.37 - 5.47 (1} _{H), 7.07 - 7.39 (2 H), 7.49 - 7.57 (1 H), 7.66 - 7.74 (1 H), 7.80 - 7.86 (1 H), 8.55 - 8.62 (1 H),} _{9.06 - 9.15 (1 H), 9.37 - 9.43 (1 H).} 10 A_{cOH (11 µL, 186 µmol) is added to a mixture of intermediate 71 (80 mg, 186 µmol) and (3S)-3-} _{15 methylpyrrolidin-3-ol 13 (40 mg, 279 µmol) in THF (1.5 mL) and the mixture is stirred for 16 h at rt.} 124 12-0519-WO-1 Sodium triacetoxyborohydride (158 mg, 744 µmol) is added and stirring is continued for 2 h at rt. The crude mixture is directly purified by prep. RP-HPLC (basic conditions) to give example E59. Example E59 Analytical HPLC-MS Method: D R_{[min]: 0.9 +} _{t 6 MS [m/z]: 516 [M+H]} Chiral SFC Method: AT R_{t [min]: 0.65 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.21 - 1.41 (5 H), 1.53 (3 H), 1.61 - 1.79 (4 H), 1.93 -} _{2.19 (5 H), 2.67- 2.75 (1 H) , 3.05 - 3.15 (1 H), 4.44 (1 H), 5.42 (1 H), 7.03 - 7.41 (2 H), 7.53 (1} H), 7.70 (1 H), 7.84 (1 H), 8.58 (1 H), 9.11 (1 H), 9.40 (1 H), missing proton(s) presumably hidden by/ overlapping with solvent signals. 5 A_{cOH (13 µL, 232 µmol) is added to a mixture of intermediate 71 (100 mg, 232 µmol) and [(2R,4R)‐} _{4‐fluoropyrrolidin‐2‐yl]methanol hydrochloride 72 (56 mg, 349 µmol) in THF (1.9 mL) and the mixture} is stirred for 16 h at rt. Sodium triacetoxyborohydride (197 mg, 929 µmol) is added and stirring is10 continued for 2 h at rt. The crude mixture is directly purified by prep. RP-HPLC to give example E60. Example E60 Analytical HPLC-MS Method: D R_{[min]: 0.94 MS [m/z +} _{t ]: 534 [M+H]} Chiral SFC Method: AK R_t [min]: 1.06 d.e. > 98% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.31 - 1.56 (5 H), 1.64 - 2.19 (8 H), 2.62 - 2.88 (2 H),} _{2.95 (1 H), 3.04 - 3.23 (3 H), 3.41 - 3.50 (1 H), 4.41 (1 H), 5.02 - 5.26 (1 H), 5.42 (1 H), 7.08 -} 7.39 (2 H), 7.53 (1 H), 7.70 (1 H), 7.84 (1 H), 8.58 (1 H), 9.12 (1 H), 9.41 (1 H). 125 12-0519-WO-1 7_{1 trans-73 trans-E61a trans-E61b} _{A mixture of intermediate 71 (150 mg, 348 µmol) and trans‐4‐fluoropiperidin‐3‐ol hydrochloride trans-} _{5 73 (114 mg, 697 µmol) in AcOH (100 µL, 1.75 mmol) and isopropanol (2 mL) is stirred for 10 min at} rt.2-Picoline-borane complex (58 mg, 523 µmol) is added, and the reaction mixture is stirred for 2 days at rt. An aqueous NaOH solution (4 M) is added, and the mixture is diluted with MeOH, filtered, and directly purified via RP-HPLC (basic conditions) and chiral SFC to obtain examples trans-E61a and t_{rans-61b as single stereoisomers. The absolute configuration of the fluorine and hydroxy substituents}10 is not known; their relative configuration is trans. Example trans-E61a Analytical HPLC-MS Method: D R_{[min]: 0.95 M +} _{t S [m/z]: 534 [M+H]} Analytical SFC method: R R_{t [min]: 3.49 d.e. > 89%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.37 - 1.61 (6 H), 1.65 - 1.79 (2 H), 1.84 - 1.93 (2 H),} _{1.94 - 2.15 (4 H), 2.26 (1 H), 2.38 - 2.49 (1 H), 2.74 - 2.84 (1 H), 2.86 - 2.95 (1 H), 3.03 - 3.13 (1} _{H), 3.41 - 3.54 (1 H), 4.00 - 4.32 (1 H), 5.11 (1 H), 5.42 (1 H), 7.07 - 7.37 (2 H), 7.53 (1 H), 7.70} (1 H), 7.83 (1 H), 8.58 (1 H), 9.11 (1 H), 9.41 (1 H). Example trans-E61b Analytical HPLC-MS Method: D R_{[min]: 0.95 MS [m/z] +} _{t : 534 [M+H]} Analytical SFC method: R R_{t [min]: 4.40 d.e. > 94%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.35 - 1.62 (6 H), 1.64 - 1.79 (2 H), 1.83 - 1.92 (2 H),} _{1.94 - 2.14 (4 H), 2.26 (1 H), 2.39 - 2.49 (1 H), 2.75 - 2.83 (1 H), 2.85 - 2.94 (1 H), 3.03 - 3.14 (1} _{H), 3.41 - 3.54 (1 H), 4.07 - 4.33 (1 H), 5.11 (1 H), 5.42 (1 H), 7.07 - 7.38 (2 H), 7.55 (1 H), 7.70} (1 H), 7.83 (1 H), 8.58 (1 H), 9.11 (1 H), 9.41 (1 H). 126 12-0519-WO-1 7_{1 74 E62} _{A mixture of intermediate 71 (140 mg, 325 µmol), (3R,5R)-5-methylpyrrolidin-3-ol trifluoroacetate 74} 5 (119 mg, 553 µmol), AcOH (38 µL, 651 µmol), and 2-picoline-borane complex (45 mg, 423 µmol) in MeOH (4 mL) is stirred for 3 days at rt. The mixture is neutralized with aqueous NaOH solution (1 N) and diluted with MeOH. The mixture is separated via prep. RP-HPLC (basic conditions) to give example E62. Example E62 Analytical HPLC-MS Method: D R_{t [min]: 0.96 MS [m/z]: 516 [M+H]}+ Analytical SFC method: AJ R_{t [min]: 4.83 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.98 (3 H), 1.32 - 1.56 (6 H), 1.64 - 2.07 (7 H), 2.41 - 2.47} _{(1 H), 2.60 - 2.70 (1 H), 2.98 - 3.14 (3 H), 4.10 - 4.18 (1 H), 4.59 (1 H), 5.43 (1 H), 7.09 - 7.38 (2 H),} _{7.53 (1 H), 7.70 (1 H), 7.83 - 7.87 (1 H), 8.58 (1 H), 9.12 (1 H), 9.40 (1 H).} 10 Synthesis of intermediate 75 A_{n aqueous HCl solution (4 M; 2 mL, 8 mmol) is added to intermediate 70 (690 mg, 1.46 mmol) in} THF (10 mL) and the mixture is stirred at rt for 16 h. An aqueous NaOH solution (4 M; 1.8 mL) and 15 saturated NaHCO3 solution is added, and the mixture is extracted with EtOAc. The combined organic layers are washed with aqueous saturated NaCl solution, and the solvent is evaporated under reduced pressure. The material is purified by column chromatography (SiO₂; EtOAc/cyclohexane: 80:20) to o_{btain intermediate 75.} 127 12-0519-WO-1 Intermediate 75 HPLC-MS Method: E R [min]: 0.97 MS [m/z + t ]: 429 [M+H] 5_{AcOH (100 µL, 1.7 mmol) and 2-picoline-borane complex (39 mg, 350 µmol) are added to a mixture} _{of intermediate 75 (100 mg, 233 µmol) and (S)-3-hydroxypiperidine hydrochloride 30 (66 mg, 467} µmol) in MeOH (3 mL). The resulting mixture is stirred for 16 h at rt. Additional 2-picoline-borane complex (20 mg) is added, and the reaction mixture is stirred for 2 days at rt. An aqueous NaOH solution (4 M, 400 µL) is added, and the mixture is diluted with MeOH, filtered, and directly purified by prep. 10 RP-HPLC (basic conditions) to obtain example E63. Separation of the diastereomers is possible by standard purification methods. Example E63 Analytical HPLC-MS Method: D R_{t [min]: 0.95 MS [m/z]: 514 [M+H]}+ 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.01 - 1.15 (1 H), 1.32 - 1.47 (1 H), 1.49 - 1.68 (5 H),} _{1.76 - 1.87 (1 H), 1.90 - 2.19 (3 H), 2.24 - 2.46 (2 H), 2.55 - 2.81 (4 H), 2.85 - 2.98 (1 H), 3.39 -} _{3.52 (1 H), 4.51 - 4.56 (1 H), 5.38 - 5.48 (1 H), 7.08 - 7.40 (3 H), 7.50 - 7.57 (1 H), 7.67 - 7.74 (1} _{H), 7.88 - 7.93 (1 H), 8.59 - 8.65 (1 H), 9.12 - 9.20 (1 H), 9.38 - 9.44 (1 H).} ₁₅ A_{cOH (49 µl, 836 µmol) is added to a mixture of intermediate 71 (120 mg, 279 µmol) and 3-} _{azabicyclo[3.1.0]hexan-1-ol hydrochloride 76 (76 mg, 558 µmol) in dimethyl sulfoxide (1 mL) and the} mixture is stirred for 10 min at rt. 2-Picoline-borane complex (49 mg, 446 µmol) is added, and the reaction mixture is stirred at rt for 16 h. The mixture is neutralized with aqueous NaOH solution (1 N), 128 12-0519-WO-1 diluted with ACN, filtered, and directly purified by prep. RP-HPLC (basic conditions) and chiral SFC t_{o obtain example E64a and E64b as single stereoisomers. The absolute configuration of the bridged} carbon atoms is not known; the relative configuration of the [3.1.0] ring system is cis. Example E64a Analytical HPLC-MS Method: D R_{t [min]: 0.94 MS [m/z]: 514 [M+H]}+ Analytical SFC method: AE R_{t [min]: 1.11 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.66 (1 H), 0.79 (1 H), 1.17 - 1.35 (3 H), 1.53 (3 H),} _{1.63 - 1.79 (2 H), 1.92 - 2.06 (4 H), 2.15 - 2.25 (1 H), 2.82 (1 H), 3.01 - 3.13 (2 H), 5.42 (1 H),} _{5.55 (1 H), 7.09 - 7.38 (2 H), 7.53 (1 H), 7.70 (1 H), 7.83 (1 H), 8.58 (1 H), 9.12 (1 H), 9.40 (1} H), missing proton(s) presumably hidden by/ overlapping with solvent signals. Example E64b Analytical HPLC-MS Method: D R_{[min]: 0.94 M +} _{t S [m/z]: 514 [M+H]} Analytical SFC method: AE R_{t [min]: 1.37 d.e. > 96%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.66 (1 H), 0.79 (1 H), 1.17 - 1.34 (3 H), 1.53 (3 H),} _{1.64 - 1.79 (2 H), 1.91 - 2.05 (4 H), 2.14 - 2.24 (1 H), 2.45 - 2.49 (2 H), 2.82 (1 H), 3.01 - 3.14 (2} _{H), 5.42 (1 H), 5.54 (1 H), 7.08 - 7.38 (2 H), 7.53 (1 H), 7.70 (1 H), 7.83 (1 H), 8.58 (1 H), 9.12} (1 H), 9.40 (1 H). 5 129 12-0519-WO-1 5_{69 77} Tert-butanole (103 mL) is added to AD-Mix beta (43 g, 55 mmol) in water (80 mL) and the mixture is stirred 20 min at rt. The mixture is cooled to 0°C. Methanesulfonamide (1.05 g, 11.1 mmol) and a s_{olution of intermediate 69 (5.2 g, 11.1 mmol) in THF (45 mL) are added and stirring is continued for} 3 days at rt. An aqueous Na2S2O3 solution (10%; 100 mL) is added and stirring continued for 30 min at 10 rt. EtOAc and saturated aqueous NaHCO3 solution are added, the organic layer is washed with saturated aqueous NaHCO3 solution and dried over MgSO4. After filtering and evaporation of the solvents, intermediate 77 is used for the next step without further purification. Intermediate 77 Analytical HPLC-MS Method: D R_{[min]: 0.8 +} _{t 9 MS [m/z]: 507 [M+H]} 130 Synthesis of intermediate 78

77 78

Aqueous HC1 (4 M; 19 mL) is added to a solution of intermediate 77 (5.6 g, 11.1 mmol) in THF (27 mL) and stirring continued for 48 h at rt. THF is evaporated and the solution is basified with aqueous NHs solution. EtOAc and water are added, the organic layer is separated and dried over MgSCU After filtration and evaporation, the material is purified by prep. RP-HPLC (acidic conditions) to obtain intermediate 78.

Intermediate 78

HPLC-MS Method: E

Rt [min]: 0.92 MS [m/z]: 445 [M+H]⁺

78 79

A mixture of intermediate 78 (2.6 g, 5.74 mmol) and Pd/C (10%w/w, 200 mg) in EtOAc (168 mL) is stirred under H2 atmosphere (30 psi) for 2 h at rt. The mixture is filtered, the solvent evaporated, and the residue purified by prep. RP-HPLC (acidic conditions) to obtain intermediate 79.

Intermediate 79

HPLC-MS Method: E

Rt [min]: 0.93 MS [m/z]: 447 [M+H]⁺ 12-0519-WO-1 7_{9 80 E65} _{Intermediate 79 (100 mg, 224 µmol) is dissolved in 1.8 mL THF. Then (R)-3-fluoropyrrolidine} _{5 hydrochloride 80 (42 mg, 336 µmol) and AcOH (13 µL, 224 µmol) are added and stirring is continued} for 2 h at rt. Then sodium triacetoxyborohydride (189 mg, 896 µmol) is added and the reaction mixture stirred for additional 2 h at rt. Water is added, filtered and the filtrate is directly purified by prep. RP- HPLC (basic conditions) to give example E65. Example E65 Analytical HPLC-MS Method: D R_{t [min]: 0.96 MS [m/z]: 520 [M+H]}+ Analytical SFC method: K R_{t [min]: 5.78 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.44 - 1.59 (5 H), 1.64 - 1.95 (5 H), 1.99 - 2.17 (1 H),} _{2.19 - 2.30 (1 H), 2.35 - 2.46 (1 H), 2.61 - 2.99 (5 H), 5.06 - 5.33 (2 H), 5.35 - 5.55 (1 H), 7.06 -} _{7.41 (2 H), 7.46 - 7.60 (1 H), 7.64 - 7.82 (1 H), 8.13 - 8.29 (1 H), 8.60 (1 H), 9.06 - 9.26 (1 H),} _{9.39 - 9.54 (1 H).} 10 I_{ntermediate 79 (100 mg, 224 µmol) is dissolved in THF (1.8 mL). Then (S)-3-fluoropyrrolidine} _{hydrochloride 81 (42.2 mg, 336 µmol) and AcOH (12.8 µL, 224 µmol) are added and stirring is} 15 continued for 2 h at rt. Then sodium triacetoxyborohydride (190 mg, 896 µmol) is added and the reaction mixture stirred for additional 2 h at rt. The mixture is filtered and directly purified by prep. RP- HPLC (basic conditions) to give ewxample E66. Example E66 132 12-0519-WO-1 Analytical HPLC-MS Method: D R_{[mi +} _{t n]: 0.96 MS [m/z]: 520 [M+H]} Analytical SFC method: K Rt [min]: 6.06 d.e. > 98% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.42 - 1.63 (5 H), 1.64 - 2.34 (7 H), 2.59 - 3.03 (5 H),} _{5.01 - 5.54 (3 H), 7.03 - 7.39 (2 H), 7.45 - 7.81 (2 H), 8.18 - 8.29 (1 H), 8.51 - 8.71 (1 H), 9.07 -} _{9.24 (1 H), 9.42 - 9.56 (1 H), missing proton(s) presumably hidden by/ overlapping with solvent} signals. 5 A_{mixture of an aqueous Na2CO3 solution (2 M, 7.0 mL, 14 mmol), intermediate 37 (2.0 g, 4.6 mmol)} _{and borane 68 (1.4 g, 5.1 mmol) in 1,4-dioxane (15 mL) is stirred under argon atmosphere. Catalyst II} (227 mg, 0.28 mmol) is added, and the mixture is stirred under argon for 1.5 h at 95 °C. Aqueous 10 saturated NaCl solution is added, and the mixture is extracted with EtOAc. The combined organic layers 133 12-0519-WO-1 are washed with saturated NaCl solution, dried over Na₂SO₄, filtered and the solvent is evaporated. The remaining material is purified by column chromatography (SiO₂; EtOAc/cyclohexane: 50:50 à 100:0) to obtain intermediate 82. Intermediate 82 Analytical HPLC-MS Method: D R [min] + t : 1.04 MS [m/z]: 491 [M+H] 5 I_{ntermediate 82 (2.1 g, 4.3 mmol) in MeOH (50 mL) and Pd/C (10%w/w, 200 mg) is stirred under H2} atmosphere (50 psi) for 18 h at rt. The mixture is filtered, and the solvent evaporated to obtain10 intermediate 83. The material is used in the next reaction step without further purification. Intermediate 83 Analytical HPLC-MS Method: D R_t [min]: 1.04 MS [m/z]: 493 [M+H]⁺ _{15 Intermediate 83 (2.1 g, 4.3 mmol) in THF (20 mL) is treated with aqueous HCl solution (4 M; 5.3 mL).} The mixture is stirred for 16 h at rt. Aqueous NaOH solution and aqueous saturated NaHCO3 solution are added, and the mixture is extracted with EtOAc. The separated organic layer is washed with aqueous saturated NaCl solution. The organic layer is evaporated under reduced pressure. The remaining material is purified by column chromatography (SiO2; EtOAc/MeOH: 100:0 à 80:20) to obtain20 intermediate 84. 134 12-0519-WO-1 Intermediate 84 HPLC-MS Method: E R [min]: 1.01 MS [m/z]: 449 [M+ + t H] 5_{A mixture of intermediate 84 (60 mg, 134 µmol) and (S)-3-hydroxypiperidine hydrochloride 30 (50} mg, 352 µmol) in MeOH (2 mL) is prepared. AcOH (16 µL, 268 µmol) and 2-picoline-borane complex (15 mg, 134 µmol) are added. The mixture is stirred for 16 h at rt. Additional AcOH (30 µL) and 2- picoline-borane complex (15 mg, 134 µmol) are added and the mixture is stirred for 24 h at rt. (S)-3- H_{ydroxypiperidine hydrochloride 30 (20 mg, 141 µmol) and 2-picoline-borane complex (20 mg, 179} 10 µmol) are added, and the mixture is stirred for 24 h at rt. An aqueous NaOH solution (4 M; 230 µL) and MeOH are added. The mixture is filtered and directly purified by reversed phase (basic conditions) to obtain example E67. Example E67 Analytical HPLC-MS Method: D Rt [min]: 1.00 MS [m/z]: 534 [M+H]⁺ Analytical SFC method: Y R_{t [min]: 3.28 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.99 - 1.12 (1 H), 1.31 - 1.52 (3 H), 1.52 - 1.58 (3 H),} _{1.58 - 1.85 (4 H), 1.85 - 1.94 (2 H), 1.94 - 2.07 (3 H), 2.09 - 2.19 (1 H), 2.39 - 2.47 (1 H), 2.64 -} _{2.73 (1 H), 2.84 - 2.91 (1 H), 3.03 - 3.13 (1 H), 3.40 - 3.48 (1 H), 4.33 - 4.71 (1 H), 5.39 - 5.48 (1} _{H), 7.37 - 7.45 (1 H), 7.65 - 7.72 (1 H), 7.80 - 7.88 (2 H), 8.56 - 8.61 (1 H), 9.13 - 9.22 (1 H),} _{9.38 - 9.44 (1 H).} 135 12-0519-WO-1 Synthesis of example E68 8_{4 35 E68} _{To a mixture of intermediate 84 (95 mg, 212 µmol) and (R)-3-hydroxypiperidine hydrochloride 35 (59} 5 mg, 424 µmol) in MeOH (2 mL), AcOH (100 µL, 1.72 mmol) and 2-picoline-borane complex (25 mg, 222 µmol) are added. The mixture is stirred for 16 h at rt. (R)-3-Hydroxypiperidine hydrochloride 35 (20 mg, 141 µmol) and 2-picoline-borane complex (20 mg, 179 µmol) are added, and the mixture is stirred for 24 h at rt. The mixture is neutralized with an aqueous NaOH solution (4 M; 400 µL) and MeOH is added. The solids are filtered off and the filtrate is purified by prep. RP-HPLC (basic10 conditions) to obtain example E68. Example E68 Analytical HPLC-MS Method: D Rt [min]: 1.00 MS [m/z]: 534 [M+H]⁺ Analytical SFC method: S R_{t [min]: 7.04 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.98 - 1.13 (1 H), 1.30 - 1.51 (3 H), 1.51 - 1.58 (3 H),} _{1.58 - 1.84 (4 H), 1.84 - 1.93 (2 H), 1.93 - 2.06 (3 H), 2.09 - 2.18 (1 H), 2.39 - 2.48 (1 H), 2.64 -} _{2.74 (1 H), 2.82 - 2.90 (1 H), 3.03 - 3.14 (1 H), 3.41 - 3.48 (1 H), 4.40 - 4.61 (1 H), 5.37 - 5.49 (1} _{H), 7.36 - 7.47 (1 H), 7.63 - 7.74 (1 H), 7.81 - 7.88 (2 H), 8.56 - 8.61 (1 H), 9.14 - 9.21 (1 H),} _{9.37 - 9.44 (1 H).} _{15 To a mixture of intermediate 84 (95 mg, 212 µmol) and (S)-3-hydroxypyrrolidine 85 (37 mg, 424 µmol)} in MeOH (2 mL), AcOH (100 µL, 1.7 mmol) and 2-picoline-borane complex (25 mg, 222 µmol) are added. The mixture is stirred for 16 h at rt. The mixture is neutralized with an aqueous NaOH solution 136 12-0519-WO-1 (4 M) and MeOH is added. The solids are filtered off and the filtrate is purified by prep. RP-HPLC (basic conditions) to obtain example E69. Example E69 Analytical HPLC-MS Method: D Rt [min]: 0.98 MS [m/z]: 520 [M+H]⁺ Analytical SFC method: T R_{t [min]: 3.74 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.24 - 1.40 (2 H), 1.47 - 1.58 (4 H), 1.65 - 1.79 (2 H),} _{1.90 - 2.02 (3 H), 2.02 - 2.15 (3 H), 2.36 - 2.42 (1 H), 2.61 - 2.69 (1 H), 2.75 - 2.82 (1 H), 3.03 -} _{3.15 (1 H), 4.12 - 4.22 (1 H), 4.45 - 4.82 (1 H), 5.37 - 5.48 (1 H), 7.37 - 7.45 (1 H), 7.63 - 7.72 (1} _{H), 7.82 - 7.88 (2 H), 8.56 - 8.60 (1 H), 9.13 - 9.22 (1 H), 9.40 - 9.44 (1 H), missing proton(s)} presumably hidden by/ overlapping with solvent signals. 5 S_{odium triacetoxyborohydride (180 mg, 849 µmol) is added to a mixture of intermediate 84 (200 mg,} _{446 µmol) and trans‐4‐fluoropiperidin‐3‐ol hydrochloride trans-73 (88 mg, 535 µmol) in DCM. The} reaction mixture is stirred for 18 h at rt. The solvent is evaporated, aqueous NaHCO₃ solution is added, 10 and the mixture is extracted with DCM. The combined extracts are purified via prep. RP-HPLC (basic c_{onditions) and by chiral SFC to obtain example trans-E70a and trans-E70b as single stereoisomers.} The absolute configuration of the fluorine and hydroxy substituent is not known; their relative configuration is trans. Example trans-E70a Analytical HPLC-MS Method: D R_t [min]: 1.00 MS [m/z]: 552 [M+H]⁺ Analytical SFC method: AA R_t [min]: 0.88 d.e. > 98% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.35 - 1.61 (6 H), 1.65 - 1.78 (2 H), 1.89 (2 H), 1.94 - 2.12} _{(4 H), 2.26 (1 H), 2.73 - 2.83 (1 H), 2.86 - 2.96 (1 H), 3.01 - 3.15 (1 H), 3.42 - 3.54 (1 H), 4.07 - 4.29} _{(1 H), 5.11 (1 H), 5.43 (1 H), 7.41 (1 H), 7.68 (1 H), 7.81 - 7.87 (2 H), 8.58 (1 H), 9.15 (1 H), 9.42} (1 H), missing proton(s) presumably hidden by/ overlapping with solvent signals. 137 12-0519-WO-1 Example trans-E70b Analytical HPLC-MS Method: D R_t [min]: 1.00 MS [m/z]: 552 [M+H]⁺ Analytical SFC method: AA R_t [min]: 1.81 d.e. > 98% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.36 - 1.62 (6 H), 1.64 - 1.79 (2 H), 1.88 (2 H), 1.94 - 2.13} _{(4 H), 2.26 (1 H), 2.73 - 2.84 (1 H), 2.86 - 2.96 (1 H), 3.02 - 3.14 (1 H), 3.43 - 3.54 (1 H), 4.06 - 4.30} _{(1 H), 5.11 (1 H), 5.43 (1 H), 7.41 (1 H), 7.68 (1 H), 7.81 - 7.87 (2 H), 8.58 (1 H), 9.16 (1 H), 9.42} (1 H), missing proton(s) presumably hidden by/ overlapping with solvent signals. Synthesis scheme of intermediate 88 5 Tert-butanol (95 mL, 1019 mmol) is added to a mixture of AD-Mix beta (39.7 g, 51 mmol) in water (83 mL) and the mixture is stirred for 20 min at rt. The mixture is cooled to 0°C. Methanesulfonamide (970 m_{g, 10.2 mmol) and a solution of intermediate 82 (5.0 g, 10.2 mol) in THF (83 mL) are added and} 138 12-0519-WO-1 stirring is continued at rt for 16 h. An aqueous Na₂S₂O₃ solution (10%, 100 mL) is added and stirring continued for 30 min at rt. EtOAc is added, the organic layer washed with saturated aqueous NaHCO₃ solution and dried over MgSO₄. After filtering and evaporation of the solvents, the remaining mixture i_{s purified via column chromatography (SiO2; EtOAc/petroleum ether: 50/50 à 0:100) to obtain} 5 intermediate 86. Intermediate 86 Analytical HPLC-MS Method: D R_{t [min]: 0.94 MS [m/z]: 525 [M+H]}+ 10 A mixture of intermediate 86 (4.7 g, 9.0 mmol) in THF (22 mL) is treated with aqueous HCl solution (4 M; 16 mL) and stirring is continued at rt for 24 h. THF is evaporated, and the aqueous layer is extracted with EtOAc. The combined organic layers are dried with MgSO4, filtered and concentrated. T_{he intermediate 87 is used for the next reaction step without further purification.} Intermediate 87 HPLC-MS Method: E R_{t [min]: 0.97 MS [m/z]: 463 [M+H]}+ 15 A mixture of intermediate 87 (4.8 g, 10 mmol) in 1,4-dioxane (85 mL) is treated with HCl in 1,4- dioxane (4 M; 25 mL) and Pd/C (10% wt/wt 500 mg). The mixture is stirred under H₂ atmosphere (50 20 psi) for 24 h at rt. The mixture is filtered, the solvents are evaporated, and the residue purified by prep. RP-HPLC (acidic conditions) to obtain intermediate 88. 139 12-0519-WO-1 Intermediate 88 HPLC-MS Method: E R_{[min]: 0.99 +} _{t MS [m/z]: 465 [M+H]} Synthesis of example E71 5_{Intermediate 88 (123 mg, 265 µmol) in THF (2.1 mL) is treated with (S)-3-fluoropyrrolidine} _{hydrochloride 81 (50 mg, 397 µmol), AcOH (30 µL, 530 µmol) and molecular sieves 4 Å and is stirred} at 55°C for 2 h. After cooling to rt, sodium triacetoxyborohydride (225 mg, 1.06 mmol) is added and the reaction mixture stirred for 1 h at rt. The crude mixture is filtered through Celite and directly purified by prep. RP-HPLC (basic conditions) to give example E71. Example E71 Analytical HPLC-MS Method: D R_{t [min]: 1.01 MS [m/z]: 538 [M+H]}+ Analytical SFC method: I Rt [min]: 1.91 d.e. > 98% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.50 - 1.97 (10 H), 2.02 - 2.21 (1 H), 2.24 - 2.37 (1 H),} _{2.61 - 3.04 (5 H), 5.08 - 5.32 (2 H), 5.37 - 5.50 (1 H), 7.37 - 7.45 (1 H), 7.65 - 7.73 (1 H), 7.80 -} _{7.90 (1 H), 8.20 - 8.25 (1 H), 8.58 - 8.62 (1 H), 9.16 - 9.24 (1 H), 9.47 - 9.52 (1 H), missing} proton(s) presumably hidden by/ overlapping with solvent signals. 10 140 12-0519-WO-1 Synthesis of intermediate 90 5 HATU (1.56 g, 4.09 mmol) and triethylamine (2.09 mL, 14.9 mmol) are added to intermediate 12 (1.00 g, 3.72 mmol) in DMF (5 mL) and the mixture is stirred for 10 min at rt. (R)-1-(3-(1,1-Difluoroethyl)- 2_{-fluorophenyl)ethanamine hydrochloride 89 (CAS: 2569698-48-0; 949 mg, 3.90 mmol) is added and} the mixture is stirred for 16 h at rt. The mixture is diluted with water and extracted with EtOAc. The 10 combined organic layers are washed with aqueous saturated NaCl solution, dried over MgSO4 and concentrated in vacuo. The material is purified by column chromatography (SiO2; EtOAc/MeOH: 96:4 à 80:20) to obtain intermediate 90. Intermediate 90 Analytical HPLC-MS Method: D R_{[min]: 0.92 +} _{t MS [m/z]: 427 [M+H]} 141

A mixture of intermediate 90 (1.00 g, 2.34 mmol), CS2CO3 (385 mg, 1.17 mmol), piperidine (116 pL, 1.17 mmol) in 1,4-dioxane (10 mL) is stirred for 4 h at 80 °C. The mixture is diluted with EtOAc, fdtered over a pad of silica gel, rinsed with EtOAc, and concentrated in vacuo. The residue is dissolved in EtOAc and washed with water. The organic layer is concentrated under reduced pressure and purified by prep. RP-HPLC (basic conditions) to obtain intermediate 91.

Intermediate 91

HPLC-MS Method: E

Rt [min] : 1.03 MS [m/z] : 427 [M+H]⁺

A mixture of intermediate 91 (330 mg, 772 μmol), piperidin-4-one hydrochloride 8 (220 mg, 1.55 mmol) and CS2CO3 (1.01 g, 3.09 mmol) in 1,4-dioxane (6 mL) is degassed with argon. Catalyst I (26 mg, 31 μmol) is added, and the mixture is stirred under argon for 3 h at 95 °C. The mixture is diluted with water and extracted with EtOAc. The combined organic layers are washed with aqueous saturated NaCl solution, dried over MgSO4 and the solvent is evaporated under reduced pressure. The material is purified by prep. RP-HPLC (basic conditions) to obtain intermediate 92.

Intermediate 92

Analytical HPLC-MS Method: D

Rt [min] : 0.96 MS [m/z] : 446 [M+H]⁺ 12-0519-WO-1 9_{2 19 E72} _{2-Picoline borane complex (16 mg, 146 µmol) is added to a mixture of intermediate 92 (50 mg, 112} _{5 µmol), L-prolinol 19 (23 mg, 224 µmol), and AcOH (20 µL, 337 µmol) in MeOH (2 mL) and the} resulting mixture is stirred for 16 h at rt. The reaction mixture is neutralized with aqueous NaOH solution (1 N), diluted with MeOH, filtered, and purified by prep. RP-HPLC (basic conditions) to obtain example E72. Example E72 Analytical HPLC-MS Method: D R_t [min]: 0.99 MS [m/z]: 531 [M+H]⁺ Analytical SFC method: Y R_t [min]: 5.24 d.e. > 98% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.47 - 1.72 (9 H), 1.85 - 1.95 (2 H), 1.95 - 2.10 (3 H), 2.62 -} _{2.73 (1 H), 2.80 - 2.94 (4 H), 3.02 - 3.11 (1 H), 4.30 - 4.40 (3 H), 5.37 - 5.47 (1 H), 7.10 - 7.18 (1 H),} _{7.26 - 7.33 (1 H), 7.42 - 7.51 (1 H), 7.60 - 7.69 (1 H), 8.47 - 8.53 (1 H), 9.01 - 9.08 (2 H), missing} proton(s) presumably hidden by/ overlapping with solvent signals. 10 2-Picoline borane complex (16 mg, 146 µmol) is added to a mixture of intermediate 92 (50 mg, 112 µ_{mol), (R)-pyrrolidin-3-ylmethanol 43 (23 mg, 224 µmol) and AcOH (20 µL, 337 µmol) in MeOH (2} 15 mL) and the resulting mixture is stirred at rt for 16 h. The reaction mixture is neutralized with aqueous NaOH solution (1 N), diluted with MeOH, and purified by prep. RP-HPLC (basic conditions) to obtain example E73. 143 12-0519-WO-1 E_{xample E73} Analytical HPLC-MS Method: D Rt [min]: 0.96 MS [m/z]: 531 [M+H]⁺ Analytical SFC method: Y Rt [min]: 3.74 d.e. > 97% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 1.30 - 1.42 (1 H), 1.48 - 1.63 (5 H), 1.73 - 1.85 (1 H), 1.90 - 2.11} _{(5 H), 2.12 - 2.26 (2 H), 2.30 - 2.37 (1 H), 2.61 - 2.69 (1 H), 2.95 - 3.06 (2 H), 4.14 - 4.30 (2 H), 4.38 - 4.61} _{(1 H), 5.37 - 5.48 (1 H), 7.13 - 7.18 (1 H), 7.26 - 7.33 (1 H), 7.43 - 7.51 (1 H), 7.61 - 7.69 (1 H), 8.49 - 8.53} _{(1 H), 9.03 - 9.11 (2 H), missing proton(s) presumably hidden by/ overlapping with solvent signals.} Synthesis scheme of intermediate 95 5 1_{2 93} _{A mixture of intermediate 12 (1.00 g, 4.13 mmol) in DCM (7 mL) and MeOH (1.5 mL) is treated} dropwise with trimethysilyldiazomethane (0.6 M in hexane; 8.26 mL, 4.96 mmol) whereby the temperature in the reaction mixture is kept below 25°C. After complete addition, stirring is continued 10 for 1 h at rt. AcOH (0.28 mL, 4.96 mmol) is added and stirring is continued for 15 min at rt. Aqueous 2 M NaHCO3 solution is added, the aqueous layer extracted with DCM, and the combined organic layers dried with MgSO4. The mixture is filtered, and the solvent is evaporated to obtain intermediate 93, which is used in the next reaction step without further purification. 144 Intermediate 93

HPLC-MS Method: E

Rt [min]: 0.69 MS [m/z]: 256 [M+H]⁺

Synthesis of intermediate 94

A mixture of intermediate 93 (400 mg, 1.48 mmol), l-methyl-l,8-diazaspiro[4.5]decane dihydrochloride 52 (390 mg, 1.63 mmol) and CS2CO3 (1.93 g, 5.94 mmol) in 1,4-dioxane (13 mL) is degassed with argon. Catalyst I (125 mg, 148 μmol) is added, and the mixture is stirred under argon at 90°C for 16 h. The solvent is evaporated, water is added and the aqueous layer extracted with EtOAc. The organic layer is dried with MgSCE. After filtration and evaporation of the solvent, the material is purified by prep. RP-HPLC (basic conditions) to obtain intermediate 94.

Intermediate 94

Analytical HPLC-MS Method: D

Rt [min] : 0.91 MS [m/z] : 330 [M+H]⁺

A mixture of intermediate 94 (107 mg, 325 μmol) in MeOH (1.3 mL) is treated with aqueous NaOH solution (4 M; 0.24 mL, 975 μmol) and stirring is continued for 16 h at rt. After evaporation of the solvents, water and aqueous HC1 solution (4 M; 0.24 mL, 975 μmol) are added and stirring is continued for 30 min at 0°C. The reaction mixture is filtered and purified by prep. RP-HPLC (acidic conditions) to obtain the desired intermediate 95.

Intermediate 95

HPLC-MS Method: E

Rt [min] : 0.61 MS [m/z] : 314 [M+H]⁺ 12-0519-WO-1 Synthesis of example E74 A_{mixture of intermediate 95 (65 mg, 206 µmol) in THF (3.3 mL) is treated with HATU (94 mg, 247} 5 µmol) and triethylamine (116 µL, 824 µmol) and the mixture is stirred for 15 min at rt. (R)-1-(3-(1,1- D_{ifluoroethyl)-2-fluorophenyl)ethanamine hydrochloride 89 (54 mg, 227 µmol) is added and stirring is} continued for 16 h at rt. The mixture is diluted with MeOH, filtered, and the mixture is purified by prep. RP-HPLC (acidic conditions) and filtered through a carbonate cartridge. The solvent is evaporated to obtain example E74. Example E74 HPLC-MS Method: E R_{[m +} _{t in]: 0.84 MS [m/z]: 501 [M+H]} Analytical SFC method: F R_{t [min]: 1.74 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.28 - 1.38 (2 H), 1.46 - 1.56 (3 H), 1.65 - 1.86 (6 H),} _{1.96 - 2.10 (3 H), 2.18 - 2.24 (3 H), 2.65 - 2.74 (2 H), 2.80 - 2.91 (2 H), 4.29 - 4.40 (2 H), 5.37 -} _{5.48 (1 H), 7.13 - 7.19 (1 H), 7.26 - 7.34 (1 H), 7.43 - 7.50 (1 H), 7.60 - 7.68 (1 H), 8.48 - 8.53 (1} _{H), 9.02 - 9.09 (2 H).} 10 A mixture of intermediate 39 (1.5 g, 3.5 mmol), piperidin-4-one hydrochloride 8 (1.0 g, 7.0 mmol) and 15 K3PO4 (2.7 g, 12.2 mmol) in 1,4-dioxane (50 mL) is degassed with argon. Catalyst I (88 mg, 104 µmol) is added, and the mixture is stirred for 3 h at 95 °C under an argon atmosphere. The mixture is filtered, and the filter is rinsed with EtOAc. The organic solvents are evaporated under reduced pressure. The material is purified by column chromatography (SiO2; EtOAc/MeOH: 95:5 à 80:20) to obtain intermediate 96. 146 12-0519-WO-1 Intermediate 96 Analytical HPLC-MS Method: D R [min]: 0 + t .94 MS [m/z]: 450 [M+H] Synthesis of example E75 5_{96 97 E75} _{A mixture of intermediate 96 (31 mg, 70 µmol), (3R)-3-methoxypyrrolidine trifluoroacetate 97 (24 mg,} _{0.11 mmol), AcOH (30.0 µL, 0.51 mmol), and 2-picoline borane complex (7.5 mg, 70 µmol) in MeOH} (1.5 mL) is stirred for 12 h at rt. The mixture is diluted with DMF und purified by prep. RP-HPLC (basic conditions) to obtain example E75. Example E75 HPLC-MS Method: A Rt [min]: 0.80 MS [m/z]: 535 [M+H]⁺ Analytical SFC method: AH R_{t [min]: 7.22 d.e. > 98%} _{1H NMR (400 MHz, DMSO-d6 + ND4OD) δ (ppm): 1.43 - 1.58 (5 H), 1.58 - 1.69 (1 H), 1.85 - 1.98} _{(3 H), 2.12 - 2.29 (1 H), 2.39 - 2.47 (1 H), 2.52 - 2.58 (2 H), 2.79 - 2.90 (3 H), 3.11 - 3.17 (3 H), 5.28} _{- 5.37 (1 H), 6.66 - 6.80 (1 H), 7.29 - 7.39 (1 H), 7.59 - 7.66 (1 H), 7.68 - 7.75 (1 H), 8.46 - 8.56 (1} _{H), 9.14 - 9.25 (1 H), missing proton(s) presumably hidden by/ overlapping with solvent signals.} 10 A_{mixture of bromide 39 (70 mg, 154 µmol), intermediate 98 (36 mg, 170 µmol), Cs2CO3 (201 mg, 617} _{15 µmol) and catalyst I (13 mg, 15 µmol) in degassed 1,4-dioxane (1.3 mL) is stirred under argon for 16 h} 147 12-0519-WO-1 at 90 °C. Water and MeOH are added, the mixture is filtered and the filtrate is concentrated in vacuo. The residue is purified by prep. RP-HPLC (basic conditions) to obtain example E76. Example E76 Analytical HPLC-MS Method: D Rt [min]: 0.96 MS [m/z]: 477 [M+H]⁺ Analytical SFC method: T R_{t [min]: 4.92 e.e. > 94%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.47 - 1.58 (3 H), 1.71 - 1.83 (2 H), 1.92 - 2.04 (2 H), 2.19} _{- 2.29 (3 H), 2.97 - 3.07 (2 H), 3.18 - 3.27 (2 H), 4.11 - 4.26 (2 H), 5.34 - 5.49 (1 H), 6.58 - 6.68 (1} _{H), 7.35 - 7.47 (1 H), 7.64 - 7.73 (1 H), 7.75 - 7.86 (1 H), 8.58 - 8.66 (1 H), 8.97 - 9.10 (1 H), 9.26 -} 9.35 (1 H). 5 A_{mixture of intermediate 39 (125 mg, 0.275 mmol) and 1-methyl-1,8-diazaspiro[4.5]decane} _{dihydrochloride 52 (72 mg, 303 µmol) in 1,4-dioxane (2.4 mL) is degassed with argon. Cs2CO3 (359} mg, 1.10 mmol) and catalyst I (23 mg, 28 µmol) is added, and the mixture is heated for 16 h at 120°C. 10 Aqueous NaHCO3 solution is added and the mixture is extracted with EtOAc. The organic layer is dried with MgSO4. After filtration and evaporation of the solvent, the mixture is purified by prep. RP-HPLC (basic conditions) to obtain example E77. Example E77 Analytical HPLC-MS Method: D R_{[min]: 1.01 MS [m/z]: 505 [ +} _{t M+H]} Analytical SFC method: AB R_{t [min]: 6.31 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.48 - 1.58 (3 H), 1.65 - 1.88 (6 H), 2.18 - 2.26 (3 H),} _{2.59 - 2.77 (2 H), 2.82 - 2.93 (2 H), 4.39 - 4.52 (2 H), 5.35 - 5.47 (1 H), 6.77 - 6.84 (1 H), 7.37 -} _{7.48 (1 H), 7.64 - 7.73 (1 H), 7.77 - 7.86 (1 H), 8.63 - 8.69 (1 H), 9.02 - 9.11 (1 H), 9.27 - 9.34 (1} H), missing proton(s) presumably hidden by/ overlapping with solvent signals. 148 Synthesis scheme of intermediate 102 11 99

A mixture of intermediate 11 (460 mg, 2.0 mmol) in AcOH (1.7 mL, 30 mmol) is stirred at 140 °C for 5 h. The mixture is cooled to rt and poured into ice-water. The solid material is fdtered off, washed with water and dried under reduced pressure at 55 °C to obtain the intermediate 99. The material is used without further purification.

Intermediate 99

HPLC-MS Method: E

Rt [min]: 0.60 MS [m/z]: 256 [M+H]⁺ Synthesis of intermediate 100

Intermediate 99 is synthesized in analogy to intermediate 39: Intermediate 99 (27 mmol), (lR)-l-[2- fluoro-3-(trifluoromethyl)phenyl]ethan-l-amine hydrochloride 36 (27 mmol), 1-propanephosphonic anhydride (53 mmol), A-methylmorpholine (80 mmol), 70 mL ACN.

Intermediate 100

HPLC-MS Method: E

Rt [min]: 0.99 MS [m/z]: 445 [M+H]⁺

Synthesis of intermediate 101

A mixture of intermediate 100 (150 mg, 337 μmol), CS2CO3 (220 mg, 674 μmol), piperidine (57 mg, 674 μmol) in 1,4-dioxane (3 mL) is stirred for 2.5 h at 90 °C. The mixture is poured into aqueous 0.5 M KHSO4 solution and is extracted with EtOAc. The solvent is evaporated under reduced pressure and the material is used in the next reaction step without purification.

Intermediate 101

Analytical HPLC-MS Method: D

Rt [min] : 1.01 MS [m/z] : 445 [M+H]⁺ 12-0519-WO-1 1_{01 47 102} _{5 A mixture of bromide 101 (60 mg, 135 µmol), tert-butyl 1,8-diazaspiro[4.5]decane-1-carboxylate} _{hydrochloride 47 (47 mg, 162 µmol) and Cs2CO3 (154 mg, 472 µmol) and catalyst I (11 mg, 13 µmol)} in degassed 1,4-dioxane (2 mL) is stirred under argon for 16 h at 90 °C. The mixture is diluted with DMF/MeOH, filtered, and washed with DMF/MeOH. The material is purified by prep. RP-HPLC (basic conditions) to obtain intermediate 102. Intermediate 102 Analytical HPLC-MS Method: D Rt [min]: 1.18 10 I_{ntermediate 102 (68 mg, 112 µmol) in 1,4-dioxane (2 mL) is treated with 4 N HCl in 1,4-dioxane (2} 15 mL) and stirred for 3 h at rt. The solvent is evaporated under reduced pressure and the material is purified by prep. RP-HPLC (basic conditions) to obtain example E78. Example E78 Analytical HPLC-MS Method: D R_{[min]: 1.04 MS [m/z]: 505 [M +} _{t +H]} Analytical SFC method: H R_{t [min]: 3.16 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.48 - 1.77 (11 H), 2.81 - 2.89 (2 H), 3.50 - 3.65 (4 H), 5.34} _{- 5.47 (1 H), 7.08 - 7.15 (1 H), 7.37 - 7.45 (1 H), 7.63 - 7.72 (1 H), 7.78 - 7.86 (1 H), 8.88 - 8.95 (1} _{H), 9.03 - 9.10 (1 H), missing proton(s) presumably hidden by/ overlapping with solvent signals.} 151 12-0519-WO-1 1_{00 52 E79} _{A mixture of bromide 100 (200 mg, 449 µmol), 1-methyl-1,8-diazaspiro[4.5]decane dihydrochloride} 5 52 (161 mg, 674 µmol) and Cs₂CO₃ (585 g, 1.8 mmol) in 1,4-dioxane (8 mL) is degassed with argon. Catalyst I (15 mg, 18 µmol) is added, and the mixture is heated under argon for 3 h at 110 °C. The mixture is filtered, washed with EtOAc, and the solvent is evaporated under reduced pressure. The m_{aterial is purified by prep. RP-HPLC (basic conditions) to obtain example E79.} Example E79 Analytical HPLC-MS Method: D R_{t [min]: 1.00 MS [m/z]: 519 [M+H]}+ Analytical SFC method: U R_{t [min]: 3.15 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.29 - 1.37 (2 H), 1.49 - 1.61 (3 H), 1.66 - 1.86 (6 H), 2.16} _{- 2.26 (3 H), 2.65 - 2.76 (5 H), 2.79 - 2.93 (2 H), 4.32 - 4.53 (2 H), 5.34 - 5.49 (1 H), 6.74 - 6.86 (1} _{H), 7.34 - 7.48 (1 H), 7.64 - 7.74 (1 H), 7.77 - 7.86 (1 H), 8.36 - 8.49 (1 H), 8.93 - 9.09 (1 H).} 10 Synthesis of intermediate 104 A_{mixture of intermediate 99 (7.0 g, 27 mmol), (R)-1-(2-methyl-3-(trifluoromethyl)phenyl)ethanamine} 103 (CAS: 2230840-58-9; 7.4 g, 27 mmol) and 4-methylmorpholine (8.8 mL, 80 mmol) in ACN (70 15 mL) is cooled to 0°C. PPA (50%, 32 mL, 53 mmol) is added dropwise and the mixture is allowed to reach rt. The mixture is poured into ice-water, ACN is evaporated under reduced pressure and water is added to the suspension. The solids are filtered, washed with water and tert-butylmethyl ether and dried to obtain intermediate 104. The material is used without further purification. 152 12-0519-WO-1 Intermediate 104 HPLC-MS Method: E R_{t [min]: 0.99 MS [m/z]: 445 [M+H]}+ 5_{A mixture of bromide 104 (60 mg, 136 µmol), 1-methyl-1,8-diazaspiro[4.5]decane dihydrochloride 52} (49 mg, 204 µmol) and potassium phosphate (133 g, 612 µmol) in 1,4-dioxane (1 mL) is degassed with argon. Catalyst I (6 mg, 7 µmol) is added and the mixture is heated under argon for 16 h at 110 °C. The mixture is diluted with 1,4-dioxane, filtered through a thiol resin, and washed with DMF/MeOH (9:1). The mixture is directly purified by prep. RP-HPLC (basic conditions) to obtain example E80. E_{xample E80} Analytical HPLC-MS Method: D R_{[m +} _{t in]: 1.03 MS [m/z]: 515 [M+H]} Analytical SFC method: AL R_{t [min]: 2.13 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.27 - 1.40 (2 H), 1.45 - 1.55 (3 H), 1.66 - 1.87 (6 H), 2.15} _{- 2.24 (3 H), 2.62 - 2.76 (5 H), 2.79 - 2.92 (2 H), 4.33 - 4.50 (2 H), 5.35 - 5.48 (1 H), 6.74 - 6.84 (1} _{H), 7.37 - 7.46 (1 H), 7.53 - 7.63 (1 H), 7.70 - 7.79 (1 H), 8.36 - 8.46 (1 H), 8.92 - 9.02 (1 H), missing} proton(s) presumably hidden by/ overlapping with solvent signals. 10 153 12-0519-WO-1 Synthesis scheme of intermediate 111 5 A_{mixture of 1,4‐dioxaspiro[4.5]decan‐8‐one 105 (3.0 g, 18.8 mmol) and (S)-3-hydroxypiperidine} _{hydrochloride 30 (2.9 g, 20.7 mmol) in THF (30 mL) is stirred at rt for 1 h. Sodium} triacetoxyborohydride (5.2 g, 24.5 mmol) is added and the reaction mixture is stirred at rt for 16 h. The m_{ixture is concentrated in vacuo and the residue is submitted to column chromatography (SiO2;}10 (DCM/MeOH/7 N NH₃ in MeOH = 50:48:2)/DCM: 10:90 à 60:40) to obtain intermediate 106. Intermediate 106 Analytical HPLC-MS Method: D R_{[min]: 0.70 MS [m +} _{t /z]: 242 [M+H]} 154 12-0519-WO-1 1_{06 107} _{A mixture of intermediate 106 (2.66 g, 11.0 mmol) and aqueous HCl (4 N; 10 mL, 40 mmol) in acetone} 5 (20 mL) is stirred at rt for 16 h. Aqueous NaOH solution (4 N; 10 mL, 40 mmol) and saturated aqueous NaHCO3 solution (10 mL) are added, and the mixture is extracted with EtOAc. The organic layers are washed with aqueous saturated NaCl solution, dried over MgSO4, and concentrated in vacuo. The i_{ntermediate 107 is used for the next step without further purification.} Intermediate 107 Analytical HPLC-MS Method: D R_{t [min]: 0.61 MS [m/z]: 198 [M+H]}+ 10 1_{07 108} _{A mixture of intermediate 107 (2.1 g, 10.8 mmol), imidazole (1.8 g, 27.0 mmol) and tert-} _{butyldimethylsilyl chloride (2.4 g, 16.2 mmol) in DMF (20 mL) is stirred for 16 h at rt. The mixture is} 15 diluted with water and extracted with EtOAc. The combined organic layers are washed with aqueous saturated NaCl solution, dried over MgSO4, and concentrated in vacuo. The residue is purified by column chromatography (SiO2; (DCM/MeOH/7 N NH3 in MeOH (50:48:2))/DCM: 4:96 à 30:70) to give intermediate 108. Intermediate 108 Analytical HPLC-MS Method: D R_{[min]: 1. +} _{t 23 MS [m/z]: 312 [M+H]} 20 Synthesis of intermediate 109 108 109 L_{DA (1 N solution in THF; 16.3 mL, 16.3 mmol) is added to intermediate 108 (2.54 g, 8.15 mmol) in} 25 THF (30 mL) at -75°C under argon atmosphere and the resulting mixture is stirred for 2 h. A solution 155 12-0519-WO-1 of N,N-bis(trifluoromethylsulfonyl)aniline (4.08 g, 11.4 mmol) in THF (30 mL) is slowly added at - 75°C and the reaction mixture is stirred for 16 h, being allowed to warm to rt. The reaction mixture is quenched with a saturated aqueous solution of NH₄Cl (50 mL) and extracted with EtOAc. The combined organic layers are washed with aqueous saturated NaCl solution, dried over MgSO_4, and concentrated 5 in vacuo. The residue is purified by column chromatography (SiO₂; cyclohexane/EtOAc: 98:2 à 70:30) to give intermediate 109. Intermediate 109 Analytical HPLC-MS Method: D R_{t [min]: 1.35 MS [m/z]: 444 [M+H]}+ Synthesis of intermediate 110 10 1_{09 110} _{A mixture of intermediate 109 (2.64 g, 5.95 mmol), bis(pinacolato)diboron (1.51 g, 5.95 mmol) and the} potassium acetate (1.46 g, 14.9 mmol) in 1,4-dioxane (30 mL) is degassed with argon. Catalyst II (218 m_{g, 298 µmol) is added, and the mixture is stirred for 1 h at 80°C under an argon atmosphere. The} 15 solvent is evaporated, and EtOAc is added. The organic layer is washed with water and aqueous saturated NaCl solution, dried over MgSO₄, and concentrated in vacuo. The residue is dissolved in DCM and purified by column chromatography (SiO₂; DCM/MeOH: 96:4 à 85:15). The product is dissolved in ACN/water, filtered, and purified via prep. RP-HPLC (basic conditions) to give intermediate 110. I_{ntermediate 110} HPLC-MS Method: M R_{[min]: 1.25 MS [m +} _{t /z]: 422 [M+H]} 20 156 12-0519-WO-1 A_{n aqueous Na2CO3 solution (2 M; 0.77 mL, 1.55 mmol) is added to a mixture of intermediate 13 (200} _{mg, 484 µmol) and borane 110 (245 mg, 581 µmol) in 1,4-dioxane (6 mL) and the mixture is degassed} with argon. Catalyst II (20 mg, 24 µmol) is added, and the mixture is stirred for 12 h at 95 °C under an argon atmosphere. The mixture is diluted with aqueous saturated NaCl solution and extracted with 5 EtOAc. The combined organic phases are washed with aqueous saturated NaCl solution, dried over MgSO_4, and concentrated in vacuo. The material is dissolved in ACN/MeOH/water, filtered, and purified by prep. RP-HPLC (basic conditions) to give intermediate 111. Intermediate 111 Analytical HPLC-MS Method: D R_{t [min]: 1.28 MS [m/z]: 628 [M+H]}+ 10 T_{etrabutylammonium fluoride (573 µl, 573 µmol) is added to intermediate 111 (120 mg, 191 µmol) in} THF (3 mL) under an argon atmosphere and the resulting mixture is stirred at rt for 20 h. The mixture is diluted with water/ACN and purified via RP-HPLC (basic conditions) to obtain example E82. Example E82 Analytical HPLC-MS Method: D R_{[min]: +} _{t 0.91 MS [m/z]: 514 [M+H]} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.01 - 1.21 (1 H), 1.31 - 1.49 (1 H), 1.49 - 1.71 (5 H), 1.75} _{- 1.88 (1 H), 1.90 - 2.23 (3 H), 2.25 - 2.48 (2 H), 2.54 - 2.86 (4 H), 2.86 - 3.02 (1 H), 3.39 - 3.55 (1} _{H), 4.43 - 4.64 (1 H), 5.38 - 5.48 (1 H), 7.07 - 7.39 (2 H), 7.48 - 7.58 (2 H), 7.59 - 7.65 (1 H), 7.65 -} _{7.72 (1 H), 8.98 - 9.07 (1 H), 9.12 - 9.19 (1 H), 9.34 - 9.41 (1 H).} 15 157 12-0519-WO-1 E_{xample E82 (55 mg, 107 µmol) in MeOH (3 mL) is stirred in the presence of Pd/C (10%w/w, 10 mg)} 5 under an H₂ atmosphere (50 psi) for 5 h at rt. The mixture is filtered and concentrated in vacuo. The residue is dissolved in MeOH, filtered, and purified by prep. RP-HPLC (basic conditions) to obtain example E81. Example E81 Analytical HPLC-MS Method: D: Rt [min]: 0.91 MS [m/z]: 516 [M+H]⁺ Analytical SFC method: Z Rt [min]: 2.93 d.e. > 98% 1_{H NMR (400 MHz, DMSO-d6) δ (ppm): 0.99 - 1.14 (1 H), 1.33 - 1.55 (6 H), 1.58 - 1.84 (4 H), 1.86} _{- 2.19 (6 H), 2.39 - 2.48 (1 H), 2.64 - 2.73 (1 H), 2.84 - 2.92 (1 H), 3.04 - 3.16 (1 H), 3.39 - 3.48 (1} _{H), 4.51 (1 H), 5.42 (1 H), 7.08 - 7.38 (2 H), 7.51 - 7.57 (2 H), 7.68 (1 H), 9.00 (1 H), 9.08 (1 H),} 9.34 (1 H). 10 A_{mixture of intermediate 39 (150 mg, 348 µmol) and borane 112 (170 mg, 522 µmol) in 1,4-dioxane} (3 mL), MeOH (1.1 mL) and aqueous Na₂CO₃ solution (2 M; 0.55 mL, 1.11 mmol) is degassed with argon. Catalyst II (28 mg, 35 µmol) is added, and the mixture is heated 3 h under argon at 110°C. To 15 the reaction mixture water is added, the aqueous layer is extracted with EtOAc, and the combined organic layers dried with MgSO4. After filtration and evaporation of the solvent, the mixture is purified by prep. RP-HPLC (basic conditions) to obtain intermediate 113. 158 12-0519-WO-1 Intermediate 113 Analytical HPLC-MS Method: D R_{t [min]: 1.08 MS [m/z]: 534 [M+H]}+ Synthesis of intermediate 114 1_{13 114} _{5 Intermediate 113 (135 mg, 253 µmol) in 1,4-dioxane (2.2 mL) is treated with HCl solution (4 M in 1,4-} dioxane; 1.26 mL, 5.06 mmol). The mixture is stirred for 16 h at rt. The solvent is evaporated, the residue co-evaporated with toluene to give intermediate 114, which is used in the next reaction step without further purification. Intermediate 114 Analytical HPLC-MS Method: D R_{[min]: 0.91 MS [m/z]: 434 [M+H +} _{t ]} 10 Synthesis of example E83 1_{14 E83} _{A mixture of intermediate 114 (135 mg, 287 µmol) in DMF (1.2 mL) is treated with acetone (73 µL,} 1.01 mmol), AcOH (16 µL, 287 µmol), and sodium triacetoxyborohydride (244 mg, 1.15 mmol). The 15 mixture is stirred for 2 h at rt. Additional acetone (73 µL, 1.01 mmol) is added and stirring is continued for 1 h at rt. Water is added, the mixture is neutralized with 10% aqueous NH3 solution, diluted with THF and MeOH and filtered. The mixture is purified by prep. RP-HPLC (basic conditions) to obtain example E83. Example E83 Analytical HPLC-MS Method: D R₊ _{t [min]: 1.02 MS [m/z]: 476 [M+H]} 159 12-0519-WO-1 Analytical SFC method: AH R_{t [min]: 6.68 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.97 - 1.12 (6 H), 1.49 - 1.59 (3 H), 2.57 - 2.85 (5 H),} _{5.37 - 5.49 (1 H), 7.37 - 7.46 (1 H), 7.59 - 7.73 (3 H), 7.78 - 7.87 (1 H), 9.00 - 9.08 (1 H), 9.13 -} _{9.22 (1 H), 9.36 - 9.42 (1 H), missing proton(s) presumably hidden by/ overlapping with solvent} signals. 160 12-0519-WO-1 Synthesis of intermediate 115 A_{mixture of intermediate 99 (2.0 g, 7.6 mmol), benzylamine 6 (CAS: 1389852-29-2; 1.8 g, 8.0 mmol),} 5 4-methylmorpholine (2.7 g, 27 mmol) in ACN (20 mL) is cooled to 0 °C. 1-Propanephosphonic anhydride (50%, 5.0 mL, 8.4 mmol) is added dropwise and the mixture is stirred for 16 h at rt. The mixture is poured into ice-water and treated with aqueous HCl solution. The solid material is filtered o_{ff, washed with water and dried in vacuo at 55 °C to obtain intermediate 115.} Intermediate 115 Analytical HPLC-MS Method: D R [min]: 0.90 MS [m/z] + t : 427 [M+H] 10 Synthesis of intermediate 116 A_{mixture of intermediate 115 (2.6 g, 6.1 mmol), Cs2CO3 (2.0 g, 6.1 mmol), piperidine (603 µL, 6.1} 15 mmol) in 1,4-dioxane (16 mL) is stirred for 7 h at 120 °C and for 48 h at rt. The solvent is evaporated under reduced pressure. EtOAc and water are added. The aqueous layer is separated and extracted with EtOAc. The combined organic layers are dried over MgSO4. The solids are filtered, and the solvent is evaporated under reduced pressure. The material is purified by prep. RP-HPLC (basic conditions) to obtain intermediate 116. Intermediate 116 Analytical HPLC-MS Method: D R₊ _{t [min]: 0.95 MS [m/z]: 427 [M+H]} 20 161 12-0519-WO-1 Synthesis of intermediate 117 A_{mixture of intermediate 116 (800 mg, 1.8 mmol), aqueous Na2CO3 solution (2 M; 2.8 mL, 5.7 mmol),} _{5 and borane 68 (575 mg, 2.1 mmol) in 1,4-dioxane (10 mL) is stirred under argon atmosphere. Catalyst} II (87 mg, 107 µmol) is added, and the mixture is stirred at 95 °C for 2 h. The mixture is diluted with water and extracted with EtOAc. The combined organic layers are washed with aqueous saturated NaCl solution, dried over Na₂SO₄, filtered and the solvents are evaporated. The material is purified by column chromatography (SiO₂; EtOAc/cyclohexane: 50:50 à 100:0) to obtain intermediate 117. Intermediate 117 HPLC-MS Method: E R [min]: 1.03 MS [m/z]: 487 + t [M+H] 10 Synthesis of intermediate 118 I_{ntermediate 117 (940 mg, 1.9 mmol) in MeOH (10 mL) is stirred under H2 atmosphere (50 psi) in the} 15 presence of Pd/C (10 %w/w, 100 mg) for 18 h at rt. The mixture is filtered, and the solvent is evaporated to obtain intermediate 118. The material is used in the next reaction step without further purification. Intermediate 118 HPLC-MS Method: E R₊ _{t [min]: 1.02 MS [m/z]: 489 [M+H]} 162 12-0519-WO-1 Synthesis of intermediate 119 I_{ntermediate 118 (783 mg, 1.6 mmol) in THF (10 mL) is treated with aqueous HCl solution (4 M; 2.0} 5 mL). The mixture is stirred for 24 h at rt. A saturated NaHCO3 solution is added, and the mixture is extracted with EtOAc. The separated organic layer is extracted with aqueous saturated NaCl solution. The solvent of the combined organic layers is evaporated under reduced pressure to obtain intermediate 119. The material is used in the next reaction step without further purification. Intermediate 119 HPLC-MS Method: E R_t [min]: 0.97 MS [m/z]: 445 [M+H]+ 10 A_{mixture of intermediate 119 (50 mg, 112 µmol) and pyrrolidine 120 (19 µL, 225 µmol) in MeOH (1} mL) is prepared and AcOH (60 µL, 1.03 mmol) is added followed by 2-picoline-borane complex (12 15 mg, 112 µmol). The mixture is stirred for 60 h at rt. An aqueous NaOH solution is added and directly purified by prep. RP-HPLC (basic conditions) to obtain example E84. Example E84 Analytical HPLC-MS Method: D Rt [min]: 1.07 MS [m/z]: 500 [M+H]⁺ Analytical SFC method: Y R_{t [min]: 5.57 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.27 - 1.41 (2 H), 1.49 - 1.56 (3 H), 1.62 - 1.77 (6 H),} _{1.92 - 2.02 (2 H), 2.03 - 2.14 (3 H), 2.99 - 3.11 (1 H), 5.37 - 5.47 (1 H), 7.07 - 7.37 (2 H), 7.49 -} 163 12-0519-WO-1 7_{.56 (1 H), 7.65 - 7.72 (1 H), 7.79 - 7.83 (1 H), 9.04 - 9.09 (1 H), 9.22 - 9.28 (1 H), missing} proton(s) presumably hidden by/ overlapping with solvent signals. 5_{A mixture of intermediate 119 (180 mg, 405 µmol) and (S)-3-hydroxypiperidine hydrochloride 30 (115} mg, 810 µmol) in iPrOH (3 mL) is prepared and AcOH (500 µL, 8.6 mmol) is added followed by 2- picoline-borane complex (45 mg, 405 µmol). The mixture is stirred for 18 h at rt. An aqueous NaOH solution (4 M; 2 mL) is added and directly purified by prep. RP-HPLC (basic conditions) to obtain example E85. Example E85 Analytical HPLC-MS Method: D R_{[min]: +} _{t 0.97 MS [m/z]: 530 [M+H]} Analytical SFC method: N R_{t [min]: 3.05 d.e. >98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 0.99 - 1.12 (1 H), 1.30 - 1.92 (13 H), 1.93 - 2.05 (3 H),} _{2.08 - 2.17 (1 H), 2.38 - 2.47 (1 H), 2.64 - 2.73 (1 H), 2.83 - 2.90 (1 H), 2.98 - 3.08 (1 H), 3.38 -} _{3.48 (1 H), 4.47 - 4.52 (1 H), 5.37 - 5.46 (1 H), 7.06 - 7.37 (2 H), 7.49 - 7.56 (1 H), 7.65 - 7.72 (1} _{H), 7.78 - 7.82 (1 H), 9.03 - 9.10 (1 H), 9.24 - 9.28 (1 H), missing proton(s) presumably hidden} by/ overlapping with solvent signals. 10 Synthesis of intermediate 122 HATU (864 mg, 2.27 mmol) and triethylamine (0.87 mL, 6.20 mmol) are added to intermediate 12115 (500 mg, 2.07 mmol) in THF (33 mL). The mixture is stirred for 15 min at rt then (1R)-1-[2-fluoro-3- 164 12-0519-WO-1 (trifluoromethyl)phenyl]ethan-1-amine hydrochloride 36 (CAS: 2230840-52-3; 503 mg, 2.07 mmol) is added. Stirring is continued for 16 h at rt. Semi-saturated aqueous NaHCO₃ solution is added, and the aqueous layer extracted with EtOAc. The combined organic layers are dried with MgSO₄. After filtration and evaporation of the solvent, the material is purified by prep. RP-HPLC (basic conditions) 5 to obtain intermediate 122. Intermediate 122 Analytical HPLC-MS Method: D R_{[min]: 1.00 MS [m/z]: 4 +} _{t 31 [M+H]} Synthesis of example E86 _{10 A mixture of intermediate 122 (70 mg, 154 µmol) and 1-methyl-1,8-diazaspiro[4.5]decane} dihydrochloride 52 (41 mg, 170 µmol) in 1,4-dioxane (1.3 mL) is degassed with argon. Cs₂CO₃ (201 mg, 617 µmol) and catalyst I (8 mg, 9 µmol) are added, and the mixture is stirred at 120 °C for 16 h. The reaction mixture is diluted with DMF/MeOH/THF, filtered, concentrated, and purified by prep. RP-HPLC (basic conditions) to obtain example E86. Example E86 Analytical HPLC-MS Method: D R_{[min] +} _{t : 1.05 MS [m/z]: 505 [M+H]} Analytical SFC method: L R_{t [min]: 1.91 e.e. > 98%} _{1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.37 - 1.48 (2 H), 1.50 - 1.58 (3 H), 1.66 - 1.94 (6 H),} _{2.24 - 2.35 (3 H), 2.69 - 2.99 (4 H), 4.01 - 4.15 (2 H), 5.39 - 5.49 (1 H), 6.86 - 6.95 (1 H), 7.37 -} _{7.47 (1 H), 7.64 - 7.73 (1 H), 7.80 - 7.88 (1 H), 7.97 - 8.04 (1 H), 8.52 - 8.63 (1 H), 9.20 - 9.29 (1} H). 15 165 12-0519-WO-1 P_{harmacological activity - BIOLOGICAL ASSAYS AND DATA} KRAS::SOS1 alphascreen binding assay This assay can be used to examine the potency with which compounds inhibit the protein-protein interaction between SOS1 and KRAS G12C or G12D. This demonstrates the molecular mode of action 5 of compounds. Low IC50 values are indicative of high potency of the SOS1 inhibitor compound in this assay setting: Reagents: GST-tagged SOS1 (564_1049_GST_TEV_ECO) produced in-house 6xHis-Tev-K-RasG12D/G12C(1-169)Avi produced in-house 10 GDP (Sigma Cat No G7127) A_{lphaLISA Glutathione Acceptor Beads (PerkinElmer, Cat No AL109)} AlphaScreen Streptavidin Donor Beads (PerkinElmer Cat No 6760002) Assay plates: Proxiplate-384 PLUS, white (PerkinElmer, Cat No 6008289) Assay buffer: 15 1 x PBS 0.1% BSA 0.05% Tween 20 KRAS::SOS1 GDP mix: 7.5 nM (final assay concentration) K-RasG12C, 10nM (final assay concentration) K-RasG12D, 10 µM 20 (final assay concentration) GDP and 5nM (final assay concentration) GST-SOS1 are mixed in assay buffer prior to use and kept at rt. Bead mix: AlphaLISA Glutathione Acceptor Beads and AlphaScreen Streptavidin Donor Beads are mixed in the dark in assay buffer at a concentration of 10 µg/mL (final assay concentration) each prior to use and kept at rt. 25 Assay protocol: Compounds are diluted to a final start concentration of 100 µM and are tested in duplicate. Assay-ready plates (ARPs) are generated using an Access Labcyte Workstation with a Labcyte Echo 550 or 555 a_{coustic dispenser. For compound a start concentration of 100 µM, 150 nL of compound solution is} transferred per well in 11 concentrations in duplicate with serial 1:5 dilutions. _{30 The assay is run using a fully automated robotic system in a darkened room below 100 Lux. 10 µL of} KRAS::SOS1 GDP mix is added into columns 1-24 to the 150 nL of compound solution (final dilution in the assay 1:100, final DMSO concentration 1 %). After a 30 minute incubation time, 5 µL of bead mix is added into columns 1-23. Plates are kept at rt in a darkened incubator. After a further 60 minutes incubation, the signal is measured using a PerkinElmer 35 Envision HTS Multilabel Reader using the AlphaScreen specifications from PerkinElmer. Each plate contains the following controls: diluted DMSO + KRAS::SOS1 GDP mix + bead mix 166 12-0519-WO-1 diluted DMSO + KRAS::SOS1 GDP mix Result calculation: IC₅₀ values are calculated and analyzed using a 4 parametric logistic model. Table 1: Inhibition values for KRAS G12C as well as for KRAS G12D. Data obtained with the disclosed a_{ssay for a selection of compounds (I) according to the invention. The value indicates an average value}5 of at least two measurements. Example IC50 [nM] IC50 [nM] Example IC50 [nM] IC50 [nM] number G12C G12D number G12C G12D E_{1 33 30 E43b 34 35} _{E2 55 46 E44 63 42} _{E3 59 50 E45 33 30} _{E4 82 70 E46 34 31} _{E5 65 57 E47a 96 99} _{E6 36 37 E47b 41 45} _{E7 50 43 E48 69 60} _{E8 70 69 E49 49 43} _{E10 74 67 E50 49 42} _{cis-E11a 22 15 cis-E52a 59 58} _{cis-E11b 28 19 cis-E52b 71 70} _{E12 30 24 trans-E53a 24 24} _{E13 52 54 trans-E53b 33 35} _{E14 69 51 E54 39 31} _{E15 68 46 E55 69 54} _{cis-E16a 45 41 E57 65 52} _{cis-E16b 26 19 E58 32 32} _{E17a 39 28 E59 42 41} _{E17b 38 32 E60 21 20} _{E18 49 37 trans-E61a 39 41} _{E19 32 27 trans-E61b 32 32} _{E20 44 33 E62 22 15} _{cis-E21a 81 61 E63 31 27} _{cis-E21b 78 82 E64a 41 39} _{E22 48 49 E64b 35 36} _{E24 56 46 E65 90 93} _{E25 63 53 E66 74 79} _{E26 86 78 E67 33 33} _{E27 77 73 E68 36 27} _{E28 70 70 E69 28 23} _{cis-E29 56 46 trans-E70a 29 34} 167 12-0519-WO-1 Example IC50 [nM] IC50 [nM] Example IC50 [nM] IC50 [nM] number G12C G12D number G12C G12D E_{30 59 51 trans-E70b 36 42} _{E31 33 28 E71 79 79} _{E32 44 43 E72 33 32} _{E33 80 73 E73 64 64} _{E34 42 34 E74 26 21} _{E35 57 48 E75 65 60} _{E36 28 22 E76 49 36} _{E37 31 21 E77 27 22} _{E38a 66 45 E78 43 41} _{E38b 59 41 E79 20 14} _{E39 53 54 E80 37 27} _{E40 50 53 E81 25 24} _{cis-E41a 50 38 E82 20 15} _{cis-E41b 41 38 E83 30 26} _{trans-E42a 26 26 E84 38 37} _{trans-E42b 37 36 E85 37 24} _{E43a 42 40 E86 61 44} Erk phosphorylation assay ERK phosphorylation assays are used to examine the potency with which compounds inhibit the SOS1- mediated signal transduction in a KRAS mutant human cancer cell line in vitro. This demonstrates the 5 molecular mode of action of compounds by interfering with the RAS-family protein signal transduction cascade. Low IC₅₀ values are indicative of high potency of the SOS1 inhibitor compounds in this assay setting. It is observed that SOS1 inhibitor compounds demonstrate an inhibitory effect on ERK phosphorylation in a KRAS mutant human cancer cell line, thus confirming the molecular mode of action of the SOS1 inhibitor compounds on RAS-family protein signal transduction. 10 ERK phosphorylation assays are performed using the following human cell line: NCI-H358 SOS2 KO (Hofmann, Gmachl, Ramharter et al, Cancer Discov.2021, 11(1):142-15): human lung cancer with a KRAS G12C mutation; Materials used: RPMI-1640 Medium (ATCC® A10491-01™) 15 DMEM Medium (Sigma Aldrich #D6429) Fetal Bovine Serum (FBS) from HyClone (SH30084-03) 384 plates from Greiner Bio-One (781182) P_{roxiplate™ 384 from PerkinElmer Inc. (6008280)} AlphaLISA SureFire Ultra p-ERK1/2 (Thr202/Tyr204) Assay Kit (ALSU-PERK-A10K) 168 12-0519-WO-1 Acceptor Mix: Protein A Acceptor Beads from PerkinElmer (6760137M) Donor Mix: AlphaScreen Streptavidin-coated Donor Beads from PerkinElmer (6760002) Human EGF 100 µg/mL Peprotech (PNr.: AF-100-15) Complete Mini, Proteaseinhibitor Cocktail Tablets, Roche #11836170001 5 Assay setup: NCI-H358 SOS2 KO are seeded at 50000 cells per well in 60 µL of DMEM with 2 % FBS, in Greiner TC 384 plates. The cells are incubated overnight in an incubator at 37 °C and 5 % CO2 in a humidified a_{tmosphere. 60 nL compound solution (10 mM DMSO stock solution) is then added using a Beckman} Coulter Labcyte Echo 550 device. After 50 min incubation in the aforementioned incubator, 5nl EGF 10 are added to each well for a final concentration of approx.8 ng/mL using a Beckman Coulter Labcyte E_{cho 550 device and cells are incubated for another 10-15 minutes before lysis. The medium is} removed, and the cells are lysed by addition of 20 µL of 1.6-fold lysis buffer from the AlphaLISA SureFire Ultra pERK1/2 (Thr202/Tyr204) Assay Kit with added protease inhibitors. After 20 minutes o_{f incubation at rt with shaking, 6 µL of each lysate sample is transferred to a 384-well Proxiplate and} 15 analyzed for pERK (Thr202/Tyr204) with the AlphaLISA SureFire Ultra pERK1/2 (Thr202/Tyr204) A_{ssay Kit. 3 µL Acceptor Mix and 3 µL Donor Mix are added under subdued light and incubated for 2} h at rt in the dark, before the signal is measured on a Perkin Elmer Envision plate reader using 384 AlphaScreen settings for Proxiplates. Data are fitted by iterative calculation with variable hill slope. The sigmoidal curve slope is fitted using a default fitting curve to ascertain IC50 values. A minimal 20 efficacy of 35% normalized pathway modulation is set as threshold for curve fitting and consideration of resulting IC50 values. T_{able 2: Erk Phosphorylation assay. The value indicates an average value of at least two measurements.} E_{1 118 E43b 101} _{E2 92 E44 58} _{E3 155 E45 139} _{E4 336 E46 162} _{E5 324 E47a 162} _{E6 247 E47b 198} _{E7 246 E48 82} _{E8 188 E49 250} _{E10 212 E50 220} _{cis-E11a 173 cis-E52a 115} _{cis-E11b 145 cis-E52b 40} _{E12 110 trans-E53a 143} 169 12-0519-WO-1 170 12-0519-WO-1 Metabolic stability in human hepatocytes The metabolic degradation of a test compound is performed with hepatocytes in suspension. After recovery from cryopreservation, human hepatocytes are diluted in Dulbecco´s modified eagle medium (supplemented with 3.5 µg glucagon/500 mL, 2.5 mg insulin/500 mL, 3.75 mg hydrocortisone/500 mL, 5_{50% human serum) to obtain a final cell density of 1.0x106 cells/mL.Following a preincubation in a cell} culture incubator (37 °C, 10 % CO2), test compound dissolved in DMSO is mixed with the hepatocyte suspension, resulting in a final test compound concentration of 1 µM and a final DMSO concentration of 0.05 %. The cell suspension is incubated at 37°C (cell culture incubator, horizontal shaker) and samples are 10 removed from the incubation after 0, 0.5, 1, 2, 4 and 6 hours. Samples are quenched with acetonitrile (containing internal standard) and pelleted by centrifugation. The supernatant is transferred to a deep- well plate and prepared for analysis of decline of parent compound by HPLC-MS/MS. The percentage of remaining test compound is calculated using the peak area ratio (test compound/internal standard) of each incubation time point relative to the time point 0 peak area ratio. 15 The log-transformed data are plotted versus incubation time, and the absolute value of the slope obtained by linear regression analysis is used to estimate in vitro half-life (T1/2). In vitro intrinsic clearance (CLint) is calculated from in vitro T1/2 and scaled to whole liverusing a hepatocellularity of 120x10⁶ cells/g liver, a human liver per body weight of 25.7 g liver/kg as well as in vitro incubation parameters, applying the following equation: 20 CL_INTRINSIC_IN VIVO [mL/min/kg] = (CL_INTRINSIC [µL/min/10⁶ cells] x hepatocellularity [10⁶ cells/g liver] x liver factor [g/kg body weight]) / 1000 Hepatic in vivo blood clearance (CL) is predicted according to the well-stirred liver model considering an average liver blood flow (QH) of 20.7 mL/min/kg: 25 CL [mL/min/kg] = CL_INTRINSIC_IN VIVO [mL/min/kg] x hepatic blood flow [mL/min/kg] / (CL_INTRINSIC_IN VIVO [mL/min/kg] + hepatic blood flow [mL/min/kg]) Results are expressed as percentage of hepatic blood flow: QH [%] = CL [mL/min/kg] / hepatic blood flow [mL/min/kg]) _{30 Table 3: Hepatic clearance expressed as percentage of liver blood flow ( QH [%]) of exemplified} _{compounds. Data obtained with the disclosed assay for a selection of compounds (I) according to the} invention. At least one measurement otherwise average values of at least two measurements. E_{xample Number Hepatic clearance - Example Number Hepatic clearance -} QH[%] in human QH[%] in human hepatocytes hepatocytes E_{1 17 E43b 25} _{E2 15 E44 16} 171 12-0519-WO-1 _{Example Number Hepatic clearance - Example Number Hepatic clearance -} QH[%] in human QH[%] in human hepatocytes hepatocytes _{E3 24 E45 <4} _{E4 20 E46 9} _{E5 16 E47a 30} _{E6 16 E47b 28} _{E7 31 E48 27} _{E8 18 E49 23} _{E10 29 E50 15} _{cis-E11a 38 cis-E52a 7} _{cis-E11b 27 cis-E52b 14} _{E12 12 trans-E53a 45} _{E13 21 trans-E53b 28} _{E14 31 E54 12} _{E15 31 E55 <4} _{cis-E16a 32 E57 15} _{cis-E16b 24 E58 15} _{E17a 13 E59 15} _{E17b 30 E60 26} _{E18 17 trans-E61a 40} _{E19 19 trans-E61b 30} _{E20 19 E62 9} _{cis-E21a 25 E63 23} _{cis-E21b 38 E64a 16} _{E22 19 E64b 23} _{E24 22 E65 38} _{E25 17 E66 32} _{E26 8 E67 10} _{E27 9 E68 12} _{E28 23 E69 <4} _{cis-E29 24 trans-E70a 25} _{E30 18 trans-E70b 19} _{E31 13 E71 7} _{E32 23 E72 21} _{E33 15 E73 24} _{E34 10 E74 8} _{E35 6 E75 9} _{E36 10 E76 20} _{E37 25 E77 <4} 172 12-0519-WO-1 Cytochrome P450 isoenzyme inhibition assays The inhibition of the conversion of a specific substrate to its metabolite is assayed at 37°C with human l_{iver microsomes and used to determine the inhibition of cytochrome P450 isoenzymes. For the} _{5 following cytochrome P450 isoenzymes, these substrates and metabolic reactions are monitored: P450} 3A4: hydroxylation of Midazolam (MDZ). The final incubation volume contains TRIS buffer (0.1 M), MgCl2 (5 mM), a certain concentration of human liver microsomes dependent on the P450 isoenzyme measured (P4503A4: 0.1 mg/ml) and a certain concentration of the individual substrate for each isoenzyme (P4503A4: Midazolam 5 µM). 10 The effect of the test compound is determined at five different concentrations in duplicate (e.g. highest concentration 50 µM with subsequent serial 1:4 dilutions) or without test compound (high control). Following a short preincubation period, reactions are started with the cofactor (NADPH, 1mM) and stopped by cooling the incubation down to 8°C and subsequently by addition of one volume of ACN. A_{n internal standard solution - usually the stable isotope of the formed metabolite - is added after} 15 quenching of incubations. Peak area analyte (=metabolite formed) and internal standard is determined by LC-MS/MS. The resulting peak area ratio analyte to internal standard in these incubations is compared to a control activity containing no test compound. Within each of the assay runs, the IC50 of a positive control inhibitor dependent on the P450 isoenzyme measured (P4503A4: ketoconazole) is determined. The assay results are plotted against compound concentrations to calculated IC50 values20 (half maximal inhibitory concentrations) for inhibitory compounds utilizing Software IDBS E- WorkBook. 173 12-0519-WO-1 Table 4: Inhibition of P450 isoenzyme 3A4. Data obtained with the disclosed assay for a selection of _{compounds (I) according to the invention with Midazolam as substrate. At least one measurement} otherwise average values of at least two measurements. E_{xample Number CYP3A4 inhibition Example Number CYP3A4 inhibition} E_{1 >50 E43b >50} _{E2 >50 E44 >50} E3 >50 E45 >50 E_{4 >50 E46 >50} _{E5 >50 E47a >50} _{E6 >50 E47b >50} _{E7 >50 E48 >50} _{E8 >50 E49 >50} E10 >50 E50 >50 c_{is-E11a >50 cis-E52a >50} _{cis-E11b >50 cis-E52b >50} E12 >50 trans-E53a >50 E13 >50 trans-E53b >50 E14 >50 E54 >50 E15 >50 E55 >50 cis-E16a >50 E57 >50 c_{is-E16b >50 E58 >50} _{E17a >50 E59 >50} E17b >50 E60 >50 E_{18 >50 trans-E61a >50} E19 >50 trans-E61b >50 E_{20 >50 E62 >50} _{cis-E21a >50 E63 >50} cis-E21b >50 E64a >50 E22 >50 E64b >50 E_{24 >50 E65 >50} _{E25 >50 E66 >50} _{E26 >50 E67 >50} _{E27 >50 E68 >50} _{E28 >50 E69 >50} _{cis-E29 >50 trans-E70a >50} E30 >50 trans-E70b >50 E31 >50 E71 >50 174 12-0519-WO-1 Mechanism-based inhibition of CYP3A4 assay (MBI 3A4): The mechanism-based inhibition towards CYP3A4 is assayed in human liver microsomes with midazolam as substrate. The test compounds and water control (wells w/o test compound) are 5 preincubated in presence of NADPH (1 mM) with human liver microsomes (0.2 mg/mL) at a concentration of 0, 5 and 25 µM for 0, 10 and 30 min. After preincubation, the incubate is diluted 1:10 (to 0.02 mg/mL) and the substrate midazolam (15 µM) is added for the main incubation (10 min). The main incubation is quenched with ACN and the formation of hydroxy-midazolam is quantified via LC/MS-MS. The formation of hydroxy-midazolam from the 30 min preincubation relative to the 10 formation from the 0 min preincubation is used as a readout. Values of less than 100 % mean that the substrate midazolam is metabolized to a lower extent upon 30 min preincubation compared to 0 min preincubation. In general, low effects upon 30 min preincubation are desired (corresponding to values close to 100 % / not different to the values determined with water control). 15 Table 5: Mechanism-based inhibition of P450 isoenzyme 3A4 with midazolam as substrate. Data o_{btained with the disclosed assay for a selection of compounds (I) according to the invention. At least} one measurement otherwise average values of at least two measurements. E_{xample Number MBI inhibition %ctrl. Example Number MBI inhibition %ctrl. [%]} E_{1 98 E45 97} _{E2 95 E46 98} 175 12-0519-WO-1 _{Example Number MBI inhibition %ctrl. Example Number MBI inhibition %ctrl. [%]} 176 12-0519-WO-1 Preparation of other SOS1 inhibitors S_{OS1 inhibitors not of formula (I) can be prepared according to literature and tested in the assays} _{described below. For example, BI-3406 can be prepared according to Cancer Discovery (2021) 11 (1):} _{5 142–157; MRTX0902 can be prepared according to J. Med.Chem.2022,65, 9678−9690; BAY-293 can} _{be prepared according to Proc Natl Acad Sci 2019, 116(7):2551-2560.} _{Combination in vitro Proliferation Assays} _{The CellTiterGlo® assay is a measure of anti-proliferative activity of a compound. The assay} quantitatively measures levels of ATP present in a cell culture, which is proportional to the number of _{10 metabolically active cells. Low IC50 values are indicative of potent anti-proliferative activity. It is} _{observed in a HER2 mutant (HER2mut) and HER2 amplified (HER2amp) cell line that zongertinib is} _{an effective anti-proliferative agent, with IC50 values in the low nM range. In contrast, the SOS1} _{inhibitors as monotherapy show IC50 values greater than 3 µM. Synergistic effects are observed when} zongertinib and SOS1 inhibitors are combined, as evidenced by a decrease in IC₅₀ compared to _{15 zongertinib or SOS1 inhibitor as monotherapy as well as a positive HSA (Highest Single Agent) Gap} _{Score (Table 7 and Table 8).} Materials used RPMI-1640 Medium (ATCC® A10491-01™) Fetal Bovine Serum (FBS, GIBCO, Thermo Fisher Scientific Inc., 26140-079) _{20 PerkinElmer Culture plate 384-well, white, opaque, sterile, tissue-culture treated #6007680} Puromycin (Sigma-Aldrich, P9620) G_{eneticin™ Selective Antibiotic (G418 Sulfate) (GIBCO, Thermo Fisher Scientific Inc.,10131-035)} _{DMSO (AppliChem #A3672,0100)} CellTiter-Glo® 2.0 (Promega, Madison, WI, US, G9243) 25 OE-19 cell line (DSMZ-German Collection of Microorganisms and Cell Cultures GmbH ACC 700) Assay setup T_{he PC-9_YVMA cell line was generated as described in Wilding, et al. Nature Cancer 2022. PC-} 9_YVMA or OE-19 cells are seeded at 500 cells/well in 50 µL/well RPMI-1640 medium + 10% FBS 177 12-0519-WO-1 in 384-well plates (note: PC-9_YVMA cell media also contains 0.5µg/ml Puromycin+0.6mg/ml G418 Sulfate). PC-9_YVMA is a NSCLC cell line with the YVMA HER2 exon 20 insertion mutation. OE- 1_{9 is a HER2 amplified esophageal cancer cell line. The cells are incubated overnight in an incubator} _{at 37 °C and 5 % CO2 in a humidified atmosphere. The next day, compounds are added to the plate} _{5 using a Tecan D300e dispenser with the following dilution series:} Zongertinib dilution series: 1000, 200, 40, 8, 1.6, 0.32nM. SOS1 inhibitors dilution series: 3000, 1500, 750, 375, 187.5, 93.75, 46.88, 23.44, 11.72, 5.86, 2.93nM. F_{or each SOS1 inhibitor experiment, an independent zongertinib experiment was performed. This is} _{reflected in the results Tables 7 and 8 with independent zongertinib IC50 values generated per} _{10 assessment of SOS1 inhibitor. Treatments were plated out in duplicate, with 0.1% DMSO as non-treated} controls. Plates are incubated for 3 (PC-9_YVMA) or 5 (OE-19) days prior to end analysis. On day of analysis, cell proliferation was analysed by CellTiterGlo®, by adding 50 µL/well CellTiterGlo® reagent, incubating for 10 min, shaking at 300rpm at room temperature, then reading luminescence in Perkin _{15 Elmer Enspire plate reader, Luminscence 384 BIA Protocol. Data are fitted by iterative calculation with} variable hill slope. The sigmoidal curve slope is fitted using a default fitting curve to ascertain IC50 values. Synergistic effects are calculated via the fold-change in IC50 values between monotherapy and c_{ombination treatments and an assessment of HSA Gap Score.} HSA function = max (EA, EB), where E = the effect for a given compound, A or B. A positive HSA Gap 20 value indicates that the combination achieves a greater anti-proliferative effect than expected for the m_{ost active single agent. The greater the HSA Gap Score, the stronger the synergistic effect (with a} threshold of +5 to indicate synergistic effect). The HSA Gap Score is calculated by taking the highest HSA Gap cell in the dose-response matrix and calculating the average of it and surrounding cells (see Table 6). 25 178 12-0519-WO-1 Table 6: Example of HSA Gap values for PC-9_YVMA cells treated with zongertinib (concentrations a_{cross the x-axis) and BI-3406 (concentrations across the y-axis). Each cell contains the HSA Gap value} _{for a given combination (e.g. the highest Gap value is highlighted in bold, 38 for 8 nM zongertinib +} 1500 nM BI-3406). Cells making up HSA Gap Score by taking the average are shaded in grey. 5 Results Zongertinib achieves an average IC₅₀ in PC-9_YVMA and OE-19 cells of 23.0 ± 5.7 nM and 4.5 ± 0.9 nM, respectively (mean ± standard deviation). When adding the various SOS1 inhibitors in combination with zongertinib, a synergistic response was achieved, as demonstrated by average HSA Gap Score of 10 21 ± 4 and 16 ± 7 in PC-9_YVMA and OE-19 cells, respectively. For demonstration purposes, the c_{hange in zongertinib IC50 in combination with 1.5 µM SOS1 inhibitor is shown in Figure 1, Table 7} _{and Table 8. The IC50 for zongertinib in PC-9_YVMA cells decreased on average 2.3-fold to 10.3 ± 2.6} nM (mean ± standard deviation) when 1.5 µM of SOS1 inhibitor was added. In OE-19 cells, the IC50 for zongertinib decreased on average 1.5-fold to 3.2 ± 0.8 nM (mean ± standard deviation) when 1.5 15 µM of SOS1 inhibitor was added. This change in IC50 was statistically significant, as demonstrated by paired t-test analyses (Figure 1). 179 12-0519-WO-1 _{wTable 7: IC50 values of zongertinib and SOS1 inhibitors alone or in combination, fold-change to the} _{IC50 of zongertinib and HSA Gap Score in PC-9_YVMA cells. Values shown for two replicate} _{experiments (Rep 1 and Rep 2). Bottom two rows describe the average and standard deviation for each} column. 180 12-0519-WO-1 181 12-0519-WO-1 _{Table 8: IC50 values of zongertinib and SOS1 inhibitors alone or in combination, fold-change to the IC50} _{of zongertinib and HSA Gap Score in OE-19 cells. Values shown for two replicate experiments (Rep 1} and Rep 2). Bottom two rows describe the average and standard deviation for each column. 182 12-0519-WO-1 183 12-0519-WO-1 184

Claims

12-0519-WO-1 C l a i m s 1_{. Zongertinib or a pharmaceutically acceptable salt thereof for use in the treatment and/or} _{prevention of cancer, wherein zongertinib or the pharmaceutically acceptable salt thereof} _{is administered in combination with a SOS1 inhibitor.} _{2. Zongertinib or a pharmaceutically acceptable salt thereof for use according to claim 1,} wherein the SOS1 inhibitor is a compound of formula (I) , wherein _{each R1 is independently selected from the group consisting of C1-6alkyl, C1-6haloalkyl and halogen;} _{p denotes 1, 2 or 3;} R² is H, Me or Et; _{V is nitrogen (-N=) or carbon} _{W is nitrogen (-N=) or carbon} _{at least one of V and/or W is nitrogen (-N=);} _{A1, A2 and A3 are each independently selected from nitrogen (N) or carbon (=CH- or =CR3-);} and is a single or double bond; _{each R3, if present, is independently selected from the group consisting of C1-6alkyl,} C1-6alkoxy, halogen and C1-6haloalkyl; _{q denotes 0, 1 or 2;} _{ring system B is selected from C3-10cycloalkyl, C4-10cycloalkenyl, 4-13 membered heterocyclyl,} _{r denotes 0, 1, 2, 3 or 4;} each R⁴, if present, is independently selected from the group consisting of R⁵ and R⁶; each R⁵ is independently selected from the group consisting of -OR⁶, -NR⁶R⁶, halogen, -CN, -C(=O)R⁶, -C(=O)OR⁶, -C(=O)NR⁶R⁶, -NHC(=O)OR⁶ and the bivalent substituent =O; each R⁶ is independently selected from the group consisting of hydrogen, C_1-6alkyl, C_1-6alkoxy, C_1-6haloalkyl, C_3-10cycloalkyl, C_4-10cycloalkenyl, 4-11 membered heterocyclyl, 185 12-0519-WO-1 wherein the C_1-6alkyl, C_1-6alkoxy, C_1-6haloalkyl, C_3-10cycloalkyl, C_4-10cycloalkenyl and 4-11 membered _{heterocyclyl are each independently optionally substituted with one or more, identical or different R7} and/or R⁸; each R⁷ is independently selected from the group consisting of -OH, NH₂, -NHR⁸, -NR⁸R⁸, halogen, -CN, and C_1-6alkoxy; each R⁸ is independently selected from the group consisting of C_1-6alkyl, C_1-6alkoxy, C_1-6haloalkyl, C_3-10cycloalkyl, C_4-10cycloalkenyl, and 4-11 membered heterocyclyl, wherein the _{C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C3-10cycloalkyl, C4-10cycloalkenyl, and 4-11 membered} _{heterocyclyl are each independently optionally substituted with one or more, identical or different R9;} each R⁹ is -OH, halogen or C_1-6alkoxy; or a salt thereof. 3_{. Zongertinib or a pharmaceutically acceptable salt thereof for use according to claim 1,} _{wherein R2 is Me, p is 2 and R1 is independently selected from the group consisting of Me,} -CFH₂, -CF₂H, -CF₃, -CFMeH, -CFMe₂, -CF₂Me, and F. 4_{. Zongertinib or a pharmaceutically acceptable salt thereof for use according to claim 1 or 2,} _{wherein the ring} _{is selected from the group consisting of} a_{nd is optionally substituted with one or two R3.} _{5. Zongertinib or a pharmaceutically acceptable salt thereof for use according to any one of} _{claims 1 to 3, wherein each R3, if present, is C1-6alkyl.} _{6. Zongertinib or a pharmaceutically acceptable salt thereof for use according to any one of} _{claims 1 to 5 wherein ring system B is selected from the group consisting of} _, _{, ,} 186 12-0519-WO-1 187 12-0519-WO-1 and w_{herein ring system B can be attached to the compound of formula (I) and to R4, if} present, at any ring position by removal of a hydrogen atom. 7_{. Zongertinib or a pharmaceutically acceptable salt thereof for use according to any one of} claims 1 to 6, wherein each R⁴, if present, is independently selected from the group consisting of , _{, , ,} 188 12-0519-WO-1 _, 189 12-0519-WO-1 190 12-0519-WO-1 . 8_{. Zongertinib or a pharmaceutically acceptable salt thereof for use according to any one of} _{claims 1 to 7, wherein the SOS1 inhibitor is selected from the group consisting of} 191 12-0519-WO-1 192 12-0519-WO-1 E E c E c E 193 12-0519-WO-1 194 12-0519-WO-1 t t E E E E 195 12-0519-WO-1 E c E c E t t 196 12-0519-WO-1 197 12-0519-WO-1 198 12-0519-WO-1 or a pharmaceutically acceptable salt thereof. 9_{. A SOS1 inhibitor for use in the treatment and/or prevention of cancer, wherein the SOS1} _{inhibitor is administered in combination with zongertinib or a pharmaceutically acceptable} salt thereof, preferably wherein the SOS1 inhibitor is a compound of formula (I) as defined i_{n anyone of claims 2 to 8.} _{10. A method of treating and/or preventing cancer, the method comprising administering to a} patient in need thereof: -_{zongertinib or a pharmaceutically acceptable salt thereof, and} _{- a SOS1 inhibitor, preferably wherein the SOS1 inhibitor is a compound of formula (I)} _{as defined in anyone of claims 2 to 8.} _{11. Use of zongertinib or a pharmaceutically acceptable salt thereof for the manufacture of a} medicament for the treatment and/or prevention of cancer, wherein zongertinib or the pharmaceutically acceptable salt thereof is administered in combination with a SOS1 i_{nhibitor, preferably wherein the SOS1 inhibitor is a compound of formula (I) as defined in} anyone of claims 2 to 8. 1_{2. Use of a SOS1 inhibitor for the manufacture of a medicament for the treatment and/or} _{prevention of cancer, wherein the SOS1 inhibitor is administered in combination with} zongertinib or a pharmaceutically acceptable salt thereof, preferably wherein the SOS1 i_{nhibitor is a compound of formula (I) as defined in anyone of claims 2 to 8.} _{13. A pharmaceutical composition comprising:} _{- zongertinib or a pharmaceutically acceptable salt thereof, and} _{- a SOS1 inhibitor, preferably wherein the SOS1 inhibitor is a compound of formula (I)} as defined in anyone of claims 2 to 8. 1_{4. A kit comprising:} 199 12-0519-WO-1 (i) a first pharmaceutical composition comprising zongertinib or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient; and (_{ii) a second pharmaceutical composition comprising a SOS1 inhibitor, wherein the SOS1} _{inhibitor is a compound of formula (I) as defined in anyone of claims 2 to 8 and a} pharmaceutically acceptable excipient. _{15. The pharmaceutical composition as defined in claim 13 or the kit as defined in claim 14,} for use in the treatment and/or prevention of cancer. 200