Modulators of ackr3 for cardiovascular or anti-platelet therapy

Abstract

Disclosed are compounds of any one of the general formulae (Ia) to (Ig); pharmaceutical compositions comprising said compounds and their use in the treatment of diseases, especially diseases mediated by ACKR3, preferably cardiovascular diseases or platelet disorders. Furthermore, methods for the treatment of diseases are disclosed, wherein mediation, such as inhibition, modulation and / or regulation, of ACKR3 is beneficial in a human or warm-blooded animal or mammal in need of such treatment.

Description

[0001] This invention relates to a novel compound for treating and preventing diseases (especially ACKR3-mediated diseases, including but not limited to cardiovascular diseases or platelet disorders), a pharmaceutical composition comprising said compound, and a method for treating and / or preventing diseases (preferably cardiovascular diseases or platelet disorders). Technical Field

[0002] This invention relates to the field of pharmaceutically active compounds, and more particularly to the field of compounds that can be used to treat and prevent conditions affecting the heart or blood vessels or to treat and prevent abnormal platelet aggregation. Background Technology

[0003] Chemokines (also known as chemokines) are a family of signaling proteins that primarily induce cell migration. Their involvement is prominent in various types of cell movement processes, such as chemotaxis, haptotaxis, chemokinesis, haptokinesis, and transcellular migration. In addition to their role as chemotactic inducers, they are involved in a wide range of important processes, such as homeostasis and development of the immune system and inflammatory signal transduction. More than 50 human chemokines have been identified, classified into four subfamilies (C, CC, CXC, CX3C). Their corresponding chemokine receptors can be divided into two main families: conventional chemokine receptors (cCKRs) and atypical chemokine receptors (ACKRs). Although all chemokine receptors are structurally homologous to GPCRs and therefore belong to the large family of G protein-coupled receptors, only members of the cCKR family transduce signals via G proteins. ACKR is involved in various transduction molecules and functions, such as recruiting beta-arrestin in the case of atypical chemokine receptor 3 (ACKR3).

[0004] Due to their crucial roles in important physiological processes, efforts have been made to develop ligands for various chemokine receptors. With Maraviroc (the first CC chemokine receptor type 5 antagonist in HIV treatment) and Plexaflor (a CXC chemokine receptor type 4 (CXCR4) antagonist initially developed for HIV treatment) being used in some cancer patients before radiotherapy or chemotherapy to mobilize stem cells into the bloodstream, chemokine receptors have proven to be viable drug targets. ACKR3 (formerly known as CXC chemokine receptor type 7 (CXCR7)) is associated with a variety of diseases. It is expressed in a range of regions, particularly in the central nervous system, adrenal glands, endothelial cells, immune cells, and platelets. In particular, the importance of ACKR3 in cardiovascular disease (CVD) has been frequently discussed in recent scientific literature. Currently, CVD is the leading cause of death worldwide, causing approximately 17.9 million deaths annually. Therefore, new treatment strategies are in high demand. Multiple studies have shown that chemokines and ACKR3 play a key role in the development of cardiovascular disease (CVD), including the upregulation of ACKR3 expression in mice or humans suffering from atherosclerosis, stroke, myocardial infarction, and platelet dysfunction (Gawaz, M. et al. Current concepts and novel targets for antiplatelet therapy, Nature Reviews Cardiology 2023, 20, 583–599).

[0005] Enhanced surface expression of ACKR3 and its endogenous ligand stromal cell-derived factor-1 (SDF-1) (also known as CXCL12) on platelets from patients with acute coronary syndrome was described. Subsequently, the role of ACKR3 in prothrombotic events was indicated and subsequently confirmed when all-cause mortality in patients with coronary artery disease (CAD) was correlated with baseline levels of ACKR3 and CXC chemokine receptor type 4 (CXCR4). Although CXCR4 is affected by CXCL12 binding leading to G... i It mediates platelet activation, but has the opposite effect when ACKR3 is activated. ACKR3 binding promotes platelet survival and inhibits platelet activation / aggregation and thrombus formation.

[0006] Bayrak et al. developed a first-in-class ACKR3 agonist for modulating platelet aggregation and degranulation. In this way, it is possible to confirm that ACKR3 binding leads to inhibition of platelet aggregation (Bayrak, A. et al. Discovery and development of first-in-class ACKR3 / CXCR7 superagonists for platelet degranulation modulation. J. Med. Chem. 2022, 65, 13365-13384).

[0007] One object of the present invention is to provide compounds effective in inhibiting, regulating, and / or modulating ACKR3. Another object is to provide compounds having one or more significantly improved potency, functional activity, and selectivity against ACKR3. Yet another object is to target ACKR3 using small molecules. A further object is to provide ACKR3 inhibitors, modulators, and / or regulators as pharmaceuticals. A still further object is to provide ACKR3 inhibitors, modulators, and / or regulators effective for treating cardiovascular diseases, platelet disorders, and other diseases. Another object is to provide such compounds, or compositions comprising such compounds, with low toxicity to humans and / or warm-blooded animals or mammals. Summary of the Invention

[0008] The above objectives have been achieved by providing compounds having any one of general formulas (Ia) to (Ig) or their stereoisomers, enantiomers, tautomers, prodrugs, and / or pharmaceutical salts:

[0009]

[0010] in

[0011] X 1 Selected from substituted or unsubstituted cyclic groups, substituted or unsubstituted heterocyclic groups, substituted or unsubstituted fused cyclic groups, and substituted or unsubstituted fused heterocyclic groups.

[0012] X 2 Selected from -SCH2-, -CH2CH2- and bonds,

[0013] X 3 Selected from carbonyl, thiocarbonyl, and methylene.

[0014] Y 1 Selected from H and methyl,

[0015] Y2 Selected from substituted or unsubstituted phenyl, substituted or unsubstituted benzyl, tert-butyl, and tert-butoxycarbonyl.

[0016] Y 3 Selected from H, substituted or unsubstituted C1-C5 alkyl and C3-C5 cycloalkyl,

[0017] Y 4 and Y 5 Is it O, or Y? 4 Selected from H and methyl, Y 5 Selected from substituted or unsubstituted benzyl, substituted or unsubstituted benzoyl, and straight-chain or branched C1-C5 alkyl carboxylic acid ester groups.

[0018] A and B are independently selected from substituted or unsubstituted heterocyclic groups and substituted or unsubstituted fused heterocyclic groups.

[0019] C is selected from substituted or unsubstituted cyclic groups, and

[0020] n is 0-2.

[0021] As used herein, the term "cyclyl" encompasses saturated or at least partially unsaturated cyclic compounds having, for example, 3 to 8 carbon atoms, such as 3, 4, 5, 6, 7, and 8 carbon atoms. Cycyl residues also encompass aryl residues, such as phenyl. The definition of cyclyl applies equally to any group containing a cyclyl group, such as phenoxy. Cycyl groups can be linked to adjacent groups (e.g., another cyclyl residue) via carbon atom (C-bonding). Cycyl residues can also exist as fused cyclyl residues, where two or more carbon atoms (e.g., 2 carbon atoms) of the first cyclyl residue can form part of the second cyclyl residue.

[0022] As used herein, the term "heterocyclic group" refers to a cyclic residue containing one to three heteroatoms (which may be independently selected from S, O, and N). A heterocyclic group may be bonded to an adjacent group (e.g., another cyclic residue) via, for example, a carbon atom (C-bonded) or a nitrogen heteroatom (N-bonded). The definition of a heterocyclic group also applies to any group containing a heterocyclic group. Heterocyclic residues may also exist as fused heterocyclic residues, where two or more atoms of the first heterocyclic residue may form a second heterocyclic group or part of a cyclic residue. Heterocyclic groups may include thiophene, pyrrolyl, pyrazolyl, imidazolyl, pyridinyl, pyrimidinyl, azacyclic butyl, pyrrolyl, piperidinyl, piperazinyl, tetrahydroquinolinyl, and dihydroindoleyl.

[0023] A preferred heterocyclic residue is thiophene-2-yl. The thiophene-2-yl may be unsubstituted or substituted by one or two substituents, preferably one substituent, and more preferably substituted at the 5-position of the thiophene-2-yl residue.

[0024] A more preferred heterocyclic residue is a (substituted or unsubstituted phenyl)thiophene-2-yl. The substituted or unsubstituted phenyl residue may be located at the 5-position of the thiophene-2-yl residue, thereby forming an (unsubstituted or 5-substituted phenyl)thiophene-2-yl residue.

[0025] Preferred examples of substituted phenyl residues include 4-hydroxy-phenyl, 2-halo-phenyl, 3-halo-phenyl, 4-halo-phenyl, 4-methyl-phenyl, 4-tert-butyl-phenyl, 4-methoxy-phenyl, and one of the aforementioned substituted phenyl residues having an additional preferred substituent (such as 3,4-dihalo-phenyl, such as 3,4-difluoro-phenyl).

[0026] Another preferred heterocyclic residue (which can form A of compound Ib, preferably R of compound IIb) 2 The alkyl group is a substituted or unsubstituted 3,4-dihydroisoquinoline-2(1H)-yl group or a substituted or unsubstituted piperidin-1-yl group (each attached to compound Ib via an N atom). Examples of substituted 3,4-dihydroisoquinoline-2(1H)-yl groups include 3,4-dihydroisoquinoline-2(1H)-yl groups. Examples of substituted piperidin-1-yl groups include 4-(alkylcarbamoyl)piperidin-1-yl groups, wherein the alkyl group is a straight-chain or branched C1-C5 alkyl group, such as tert-butyl.

[0027] As used in this article, the term "bond" describes a chemical bond formed between two residues, particularly a covalent single bond.

[0028] As used herein, the term "alkyl" encompasses both straight-chain and branched carbon chains. Preferred alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, 2-butyl, isobutyl, tert-butyl, pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, 2,2-dimethylpropyl, and 1-ethylpropyl. The definition of alkyl also applies to any group containing an alkyl group, such as alkoxy or thioalkoxy groups.

[0029] As used herein, the term "substituted" or "substituent" may refer to a group comprising H, alkyl, heteroalkyl, cycloyl, heterocyclic, fused cycloyl, fused heterocyclic, and combinations thereof. Furthermore, the term "substituted" may refer to a group comprising: halogen or halogenated (such as F, Cl, Br, I), modification of carbon by introducing one or more heteroatoms (such as N, S, or O), or a carboxylic acid ester functional group, a primary, secondary, or tertiary amine functional group (which may optionally be further substituted), an O group with a substituent, or a carboxyl group (which may optionally be further substituted).

[0030] Preferably, “substituted” means alkyl (such as straight-chain or branched C1-C5 alkyl), hydroxy, alkoxy (such as straight-chain or branched C1-C5 alkoxy), thiol, thioalkyl (such as straight-chain or branched C1-C5 thioalkyl), halogen or halogenated (such as F, Cl, Br, I), nitro, amine, monoalkylamine or dialkylamine (where each alkyl residue is independently selected from straight-chain or branched C1-C5 alkyl), alkyl-formamide and cyclo-formamide.

[0031] Preferred examples of substituted benzyl residues include 2-halo-benzyl, 3-halo-benzyl, 4-halo-benzyl, 3-nitro-benzyl, and 4-methoxy-benzyl.

[0032] The molecular sizes of the residues described herein are chosen such that the compounds of this invention do not exhibit negative effects or even steric hindrance that hinders biological activity. Therefore, the total molecular weight of the substituents can be about ≤ 200 g / mol, such as about ≤ 150 g / mol or about ≤ 100 g / mol.

[0033] As used herein, the term "small molecule" can refer to compounds of the present invention having a low molecular weight of less than 1,000 g / mol, such as 900 g / mol to 300 g / mol, 800 g / mol to 350 g / mol, 700 g / mol to 400 g / mol, 600 g / mol to 450 g / mol, or 650 g / mol to 500 g / mol. Small molecules have the advantage of simpler structures, which facilitates their synthesis. Furthermore, their low molecular weight makes their pharmacokinetics and / or pharmacodynamics more predictable, which in turn can be advantageous for drug administration. In addition, small molecules can be readily administered (e.g., orally), and / or they can cross cell membranes to reach intracellular targets.

[0034] The terms “compound of the present invention”, “compound of the present invention”, or “compound of formula (I)” or any other formula and / or structure include its pharmaceutical salts, prodrugs, biologically active metabolites, solvates, and stereoisomers.

[0035] Pharmaceutical salts can be acid addition salts of pharmaceutical acids. Examples of suitable pharmaceutical organic and inorganic acids include hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, aminosulfonic acid, C1-C4-alkylsulfonic acids (such as methanesulfonic acid), alicyclic sulfonic acids (such as S-(+)-10-camphorsulfonic acid), aromatic sulfonic acids (such as benzenesulfonic acid and toluenesulfonic acid), dicarboxylic acids having 2 to 10 carbon atoms, and hydroxycarboxylic acids (such as oxalic acid, malonic acid, maleic acid, fumaric acid, lactic acid, tartaric acid, citric acid, glycolic acid, adipic acid, and benzoic acid). Other suitable pharmaceutical acids may be derived, for example, from Remington: The Science and Practice of Pharmacy, 23rd edition, October 30, 2020, Adeboye Adejare.

[0036] The present invention also includes any tautomer, crystalline and polymorphic forms of the said compound, its salts and mixtures thereof.

[0037] The invention also includes solvates, such as hydrates.

[0038] The compounds of the present invention may contain one or more chiral centers and exist in different optically active forms (such as enantiomers and diastereomers).

[0039] As used herein, the term "prodrug" refers to a drug that is converted into a parent drug in vivo through some physiological and chemical process. A non-limiting example of a prodrug is the compound of this invention in ester form.

[0040] Prodrugs possess many useful properties. For example, a prodrug may have higher water solubility than the final drug, thus facilitating intravenous administration. The oral bioavailability of a prodrug may also be higher than that of the final drug. After administration, the prodrug undergoes enzymatic or chemical cleavage to deliver the final drug into the blood or tissues. Suitable prodrugs are known to those skilled in the art.

[0041] The compounds of the present invention can mediate (i.e., inhibit, regulate, and / or modulate) ACKR3 and are therefore useful in therapies, particularly for treating diseases or conditions at least partially mediated by ACKR3. They may be particularly useful in treating cardiovascular diseases or platelet disorders.

[0042] Exemplary cardiovascular diseases include coronary artery disease (such as angina and heart attack), stroke, heart failure, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, arrhythmia, congenital heart disease, valvular heart disease, myitis, aortic aneurysm, peripheral artery disease, thromboembolic disease, venous thrombosis, and atherosclerosis. Exemplary platelet disorders include thrombocytosis, thrombocythemia, and thrombocytopenia (such as immune thrombocytopenia and thrombotic thrombocytopenic purpura), Bernard-Soulier disease, Glanzmann's thrombasthenia, Hermansky-Pudlak syndrome, Jacobsen syndrome, Lowe syndrome, platelet release and storage pool defects, thrombocytopenia with radius loss (TAR) syndrome, and thrombotic thrombocytopenic purpura (TTP).

[0043] The compounds of this invention can also be used to treat other diseases and conditions regulated by ACKR3, such as distress dysfunction diseases or conditions, acute or chronic pain, cancer (breast cancer, brain cancer, lymphoma), fibrosis (e.g., myocardial fibrosis), inflammatory or autoimmune diseases and conditions, conditions of excessive or abnormal angiogenesis (e.g., wound healing), stem cell differentiation and mobilization disorders, brain and neuronal dysfunction (e.g., Alzheimer's disease, multiple sclerosis, and demyelinating diseases), renal dysfunction, renal insufficiency, preeclampsia, and obesity.

[0044] According to a preferred embodiment, the compound has formula (Ia) or (Ib), wherein

[0045] X 1 It is a substituted or unsubstituted heterocyclic group, preferably a substituted or unsubstituted thiophene-2-yl, more preferably a (substituted or unsubstituted phenyl)thiophene-2-yl.

[0046] X 2 It is a key.

[0047] X 3 It is a carbonyl group.

[0048] Y 1 It's H.

[0049] Y 2 It is an unsubstituted benzyl group.

[0050] Y 3 Selected from straight-chain or branched C1-C4 alkyl, C2-C3 alkylphenyl and C3-C4 cycloalkyl, preferably methyl, and

[0051] A is selected from substituted or unsubstituted 1-piperidinyl, substituted or unsubstituted 3,4-dihydroisoquinoline-2(1H)-yl, and

[0052] n is 1.

[0053] According to another preferred embodiment, the compound has any one of general formulas (IIa) to (IIc):

[0054]

[0055] in

[0056] R 1 yes , , , , , , , , , , , , , , , or ,

[0057] R 2 yes , , , , , , , ,

[0058] , , , or ,

[0059] R 3 yes , , , , , , , , , , , or ,and

[0060] R 4 Water halogen, preferably F.

[0061] Alternatively, the compound has the general formula (IIa), wherein R 1 yes R 6 It is one or two residues independently selected from substituents, wherein the substituent is optionally further substituted. Preferably, R 6 It is one or two residues independently selected from the group consisting of: H, substituted or unsubstituted C1-C5 alkyl, halogenated (such as F, Cl, Br, I), substituent O (such as OH), and substituent (such as SH), carboxyl (which may optionally be further substituted). More preferably, R 6 It is one or two residues independently selected from the group consisting of: H, unsubstituted alkyl, straight or branched unsubstituted alkyl, OH, O (alkyl), halogenated or dihalogenated (such as F, Cl, Br, I).

[0062] Alternatively, the compound has the general formula (IIb), wherein R 2 yes R 6 Independently selected from substituents, wherein the substituent is optionally further substituted. Preferably, R 6 Independently selected from the group consisting of: H, substituted or unsubstituted C1-C5 alkyl groups, substituted or unsubstituted C3-C5 cycloalkanes, halogenated groups (such as F, Cl, Br, I), carboxylic acid ester functional groups (which may optionally be further substituted), and O groups with substituents. More preferably, R 6 Independently selected from the group consisting of: H, halogens (such as F, Cl, Br, I), unsubstituted alkyl groups, unsubstituted cycloyl groups, and O (C1-C5 unsubstituted alkyl groups). More preferably, R 6 Independently selected from the group consisting of: H, F, Cl, Br, unsubstituted alkyl, unsubstituted cycloyl, substituted or unsubstituted C1-C4 alkyl, straight-chain or branched C1-C4 alkyl, unsubstituted cycloyl, C2-C3 alkylphenyl; R at the benzyl group 6 Choose independently from the group consisting of: H, Br, F, O (alkyl), NCO (substituted cycloalginate).

[0063] According to yet another preferred embodiment, the compound has the general formula (IId) or (IIe):

[0064]

[0065] in

[0066] R 5 yes , , , or .

[0067] According to a preferred embodiment, the compound's β-repressor protein recruits EC. 50 The concentration is below 0.5 µM, preferably below 0.1 µM, and / or P-selectin expression is reduced to 71% or more, preferably 90% or more. The compound recruits EC24 as a β-repressor protein. 50 The concentration is below 0.5 µM, such as below 0.45 µM, below 0.40 µM, below 0.35 µM, below 0.30 µM, below 0.25 µM, below 0.20 µM, or below 0.15 µM. Preferably, the compound recruits EC24 as a β-repressor protein. 50 The concentration is below 0.1 µM. The expression of P-selectin in the compound is reduced to 71% or more, such as 72% or more, 73% or more, 74% or more, 75% or more, 80% or more, or 85% or more. Preferably, the expression of P-selectin in the compound is reduced to 90% or more, such as 95% or more, 96% or more, or 97% to 100%. β-inhibitor protein recruits EC. 50 Decreased expression of P-selectin can be measured according to the instructions.

[0068] According to another preferred embodiment, a pharmaceutical composition is provided comprising the compound of the present invention or its pharmaceutical salt, prodrug, biologically active metabolite, solvate or stereoisomer.

[0069] According to yet another preferred embodiment, the pharmaceutical composition further comprises an inert carrier and / or one or more other therapeutic agents.

[0070] According to a preferred embodiment, the compounds of the present invention or their pharmaceutical salts, prodrugs, biologically active metabolites, solvates or stereoisomers, or compositions of the present invention, are provided for the treatment of diseases or conditions.

[0071] According to another preferred embodiment, the disease is mediated by ACKR3.

[0072] According to another embodiment of the invention, the disease or condition is selected from the group consisting of: pain, painful functional disorders or conditions of the subject, cancer, inflammatory or autoimmune diseases and conditions, conditions of excessive or abnormal angiogenesis (e.g., wound healing and HIV infection), stem cell differentiation and mobilization disorders, brain and neuronal dysfunction (e.g., Alzheimer's disease, multiple sclerosis and demyelinating diseases), renal dysfunction, renal insufficiency, preeclampsia and obesity.

[0073] According to yet another preferred implementation, the disease is a cardiovascular disease or a platelet disorder.

[0074] According to a preferred embodiment, the cardiovascular disease is selected from coronary artery disease (such as angina pectoris and heart attack), stroke, heart failure, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, arrhythmia, congenital heart disease, valvular heart disease, myitis, aortic aneurysm, peripheral artery disease, thromboembolic disease, venous thrombosis, and atherosclerosis, and the platelet disorders are selected from thrombocytosis, thrombocytopenia, and thrombocytopenia (such as immune thrombocytopenia and thrombotic thrombocytopenic purpura), Birx-Su syndrome, Grantzmann platelet atony, H.P. syndrome, Jacobson syndrome, Loehr syndrome, platelet release and storage pool defects, thrombocytopenia with radius loss (TAR) syndrome, and thrombotic thrombocytopenic purpura (TTP).

[0075] According to another preferred embodiment, a method for treating a disease is provided, wherein the mediation of ACKR3 (such as activation or inhibition, regulation and control) is beneficial in humans or warm-blooded animals or mammals in need of such treatment, the method comprising administering to said human or warm-blooded animal or mammal a therapeutically effective amount of the compound of the invention or its pharmaceutical salt, prodrug, biologically active metabolite, solvate or stereoisomer, or composition of the invention.

[0076] According to yet another preferred implementation, the disease is a cardiovascular disease or a platelet disorder.

[0077] According to a preferred embodiment, the cardiovascular disease is selected from coronary artery disease (such as angina pectoris and heart attack), stroke, heart failure, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, arrhythmia, congenital heart disease, valvular heart disease, myitis, aortic aneurysm, peripheral artery disease, thromboembolic disease, venous thrombosis, and atherosclerosis, and the platelet disorders are selected from thrombocytosis, thrombocytopenia, and thrombocytopenia (such as immune thrombocytopenia and thrombotic thrombocytopenic purpura), Birx-Su syndrome, Grantzmann platelet atony, H.P. syndrome, Jacobson syndrome, Loehr syndrome, platelet release and storage pool defects, thrombocytopenia with radius loss (TAR) syndrome, and thrombotic thrombocytopenic purpura (TTP).

[0078] The compounds of the present invention can be administered conventionally in the form of pharmaceutical compositions comprising at least one compound according to the invention, optionally together with a carrier (such as a pharmaceutical excipient) and optionally one or more other drugs (such as drugs suitable for the treatment or prevention of cardiovascular disease or platelet disorders).

[0079] These compositions can be administered, for example, orally, rectally, transdermally, subcutaneously, intraperitoneally, intravenously, intramuscularly, or intranasally. Examples of suitable pharmaceutical compositions are solid dosage forms, such as powders, granules, tablets (especially film tablets), lozenges, sachets, capsules, sugar-coated tablets, capsules (such as hard gelatin capsules and soft gelatin capsules), or suppositories; semi-solid dosage forms, such as ointments, creams, hydrogels, pastes, or patches; and liquid dosage forms, such as solutions, emulsions (especially oil-in-water emulsions), suspensions (e.g., lotions), injectable formulations, and infusion formulations. Liposomes and / or microspheres may also be used.

[0080] In the production of these compositions, the compounds according to the invention are optionally mixed with or diluted with one or more carriers (such as pharmaceutical excipients). The carriers (such as pharmaceutical excipients) can be solid, semi-solid, or liquid materials, which serve as a medium, carrier, or medium for the compounds of the invention.

[0081] Suitable carriers (such as pharmaceutical excipients) are known to those skilled in the art and are readily available from textbooks. Formulations of suitable carriers (such as pharmaceutical excipients) may contain pharmaceutical excipients such as wetting agents; emulsifiers and suspending agents; preservatives; antioxidants; anti-irritants; chelating agents; coating agents; emulsion stabilizers; film-forming agents; gelling agents; odor masking agents; flavoring agents; resins; hydrocolloids; solvents; solubilizers; neutralizing agents; diffusion promoters; pigments; quaternary ammonium compounds; refining and overfatting agents; raw materials for ointments, creams, or oils; organosilicon derivatives; spreading agents; stabilizers; sterilizing agents; suppository bases; tablet excipients (such as binders, fillers, flow aids, disintegrants, or coatings); propellants; desiccants; opacifiers; thickeners; waxes; plasticizers; and white mineral oil. Formulations with suitable carriers can be prepared according to, for example, Fiedler, HP, Lexikon der Hilfsstoffe fur Pharmazie, Kosmetik undangrenzende Gebiete [Encyclopedia of auxiliary substances for pharmacy, cosmetics, and related fields], 5th edition, ECV-Editio-Cantor-Verlag, 2001.

[0082] The compounds of the present invention can be administered in unit doses ranging from 2.5 to 5000 mg / m² for humans or animals, particularly warm-blooded animals or mammals. 2 The dosage may be in the range of 0.05 to 100 mg / kg of body surface area. Unit dosage forms may contain 0.1 to 250 mg of the compound of the present invention. The daily dose will need to vary depending on the host being treated, the route of administration, any combination therapies, and the severity of the condition.

[0083] The compounds of the present invention can also be adapted for use in combination with other therapeutic agents or drugs. Therefore, the present invention also relates to a combination comprising the compounds of the present invention with one or more other therapeutic agents, particularly for the treatment of cardiovascular diseases, thrombocytopenia, cardiovascular disease-related diseases, or thrombocytopenia-related diseases. The combination therapies of the present invention can be administered adjunctive. Adjunctive administration means the use of each of the components in contiguous or overlapping forms as a single pharmaceutical composition or device. Such therapeutic administration regimens of two or more therapeutic agents are commonly referred to by those skilled in the art and are referred to herein as adjunctive therapeutic administration; it is also referred to as add-on therapeutic administration. Any and all treatment regimens in which a patient receives a single but contiguous or overlapping therapeutic administration of the compounds of the present invention with at least one other therapeutic agent are within the scope of the present invention. In one embodiment of adjunctive therapeutic administration as described herein, a patient typically takes one or more of these components for a period of time until stabilized, and then receives administration of another component.

[0084] The combination therapies of the present invention can also be administered simultaneously or sequentially. Simultaneous administration means a treatment regimen in which individual components are administered together, either as a single pharmaceutical composition or as a device containing or comprising two components, or as separate compositions or devices (each containing one of these components) administered simultaneously. Such combinations of separate individual components for simultaneous combined administration can be provided in the form of kit-of-parts.

[0085] Suitable pharmaceutical products for use in combination with the compounds of the present invention include, for example: anticoagulants (such as apixaban, dabigatran, edoxaban, heparin, rivaroxaban, warfarin); antiplatelet drugs and drugs used for dual antiplatelet therapy (DAPT) (such as acetylsalicylic acid, clopidogrel, dipyridamole, prasugrel, ticagrelor). elor); angiotensin-converting enzyme (ACE) inhibitors (such as benazepril, captopril, enalapril, fosinopril, lisinopril, moexipril, perindopril, quinapril, ramipril, trandolapril); angiotensin II receptor blockers (or inhibitors) Such as azilsartan, candesartan, eprosartan, irbesartan, losartan, olmesartan, telmisartan, valsartan; angiotensin receptor-neprilysin inhibitors (ARNIs) (such as sacubitril / valsartan); β-blockers (such as acetaminophen / ... butolol), atenolol, betaxolol, bisoprolol / hydrochlorothiazide, bisoprolol, metoprolol, nadolol, propranolol, sotalol); combined α- and β-receptor blockers (such as carvedilol, labetalol hydrochloride); calcium channel blockers (such as amlodipine, diltiazem, felodipine, nifedipine, nimodipine, nisoldipine, verapamil);Cholesterol-lowering drugs (such as statins, including atorvastatin, fluvastatin, lovastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin; niacin derivatives, including niacin; cholesterol absorption inhibitors, including ezetimibe; combination statins and cholesterol absorption inhibitors, such as ezetimibe / simvastatin); digitalis preparations (such as digoxin). igoxin); diuretics (such as acetazolamide, amiloride, bumetanide, chlorothiazide, chlorthalidone, furosemide, hydrochlorothiazide, indapamide, metalozone, spironolactone, torsemide); and vasodilators (such as isosorbide dinitrate, isosorbide mononitrate, hydralazine, nitroglycerin, minoxidil). Detailed Implementation

[0086] Design of novel compounds. The main objective of this invention is to identify potent ACKR3 ligands and screen them for their functional activity related to platelet regulation. Starting with the publications of Yoshikawa et al., a group of compounds derived from molecular docking studies and evaluated for their binding affinity to ACKR3 (Yoshikawa, Y. et al. Optimized method of g-protein-coupled receptor homology modeling: Its application to the discovery of novel CXCR7 ligands). J. Med. Chem. 2013, 56, 4236-4251), demonstrated small molecules with moderate to good biological activity in the ACKR3 model. One of the reported ligands was selected for optimization to obtain lower EC5 values.50 Value and functional activity. A systematic approach was chosen for the optimization process. First, the rigid cyclic portion of the reported ligand was removed to open a more flexible core structure. Figure 1 The initial compound 1 has shown a 5-fold improvement in potency in our β-inhibitor protein recruitment assay. Subsequently, this novel hit molecule was broken down into three core components: the head, tail, and linker. Figure 1 ).

[0087] Starting with the head component, each part was then derivatized and its affinity for ACKR3 was tested individually. In-depth structure-activity relationship studies yielded the lead compound 27 (EC). 50 Compound 27 (0.069 µM; P-selectin expression decreased by 77%) exhibits excellent β-inhibitor recruitment efficacy, complete agonist properties, and significant platelet functional activity. Therefore, the metabolic stability, cytotoxicity, and selectivity against ACKR3 of compound 27 were further evaluated.

[0088] The following describes the rational design, development, and evaluation of novel ACKR3 agonists that exhibit high potency, efficacy, and selectivity against ACKR3. Furthermore, they inherit excellent properties such as functional activity and low cytotoxicity, allowing this class of new compounds to be used in future in vivo studies.

[0089] Chemistry. The synthesis of the final compounds was achieved via a series of different routes, but a significant number of the final products (compounds 1–32) could be synthesized using a modular 3–5 step synthesis (Scheme 1). It begins with an in-situ Finkelstein reaction and nucleophilic substitution of (3-bromopropyl)carbamate with the corresponding alkylbenzylamine to give a Boc-protected linker as a precursor. Although most alkylbenzylamines are commercially available, the precursors of compounds 20, 29, and 30 (compounds 20c, 29c, and 30c) were first synthesized via either nucleophilic substitution or reductive amination. After successful purification, the Boc-protected linker was deprotected using a solution of 1,4-dioxane in 4 M HCl. If possible, the solid HCl salt of the free amine was collected by filtration and washed to obtain the pure product; otherwise, the solvent was evaporated, and the intermediate was used without further purification. The amine could then be coupled with a suitable carboxylic acid using HATU and DIPEA (as a base) to obtain the final compound. Compound 35 was synthesized similarly (Scheme 2), starting with commercially available tetrahydroisoquinoline. In the case of left-sided substituted compounds (“head optimization”, structure A, Table 1), multiple compounds were synthesized by applying additional steps. By first synthesizing the bromothiophene derivative (compound 2), a series of different “left-sided” substituted derivatives (compounds 9-17, 31, 32) could be obtained in a short time using a Suzuki coupling reaction with the corresponding boric acid (pinacol ester).

[0090] Scheme 1. Compounds 1-33 a Synthesis

[0091]

[0092] a Reagents and conditions: (a) Methylamine, 1-(bromomethyl)-4-fluorobenzene, Cs₂CO₃, KI, acetone, rt (room temperature), 72 h, or benzaldehyde, the corresponding amine, NaBH₃CN, EtOH, rt, 16 h, 33%-38%; (b) tert-butyl (3-bromopropyl)carbamate, the corresponding benzylamine, Cs₂CO₃, KI, acetone, rt, 72 h, 39%-69%; (c) 1,4-dioxane solution in 4M HCl, rt, 1 h, 52%-98%; (d) the corresponding carboxylic acid, DIPEA, HATU, DMF, rt, 16 h, 7%-86%; (d) the corresponding boric acid (pinacol ester), K₂CO₃, XPhos Pd G₄, water / 1,4-dioxane (1:4), 100°C, 16 h, 18%-72%.

[0093] Similar to the synthesis described above (Scheme 1), compounds 33, 38, 40, 44, 45, and 49 can be obtained in a brief one-step amide coupling reaction. Compound 33 is then further derivatized into compound 34 in two steps (Boc deprotection and reductive amination).

[0094] Although most compounds can be obtained as described in the direct manner, the remaining compounds require synthesis in multiple different steps due to their heterogeneous molecular structures. The synthesis of compound 37 begins with the saponification of N-boc-protected ethyl piperidine carboxylate. Subsequently, the resulting carboxylic acid (37e) is amide-coupled with tert-butylamine to give intermediate 37d. After obtaining the deprotected amine using a solution of 1,4-dioxane in 4M HCl, the compound is treated as described above: in-situ Finkelstein reaction with nucleophilic substitution, Boc deprotection, and final amide coupling to give compound 37.

[0095] Compound 39 was synthesized in three steps: first, by reductive amination of benzaldehyde with Boc-protected 3-methylpiperidine; then by Boc deprotection using a 1,4-dioxane solution of 4M HCl; and finally by amide coupling with a carboxylic acid.

[0096] Although compounds 36, 41, and 43 share similar structural elements, their synthesis was achieved via different routes. 36 and 41 were synthesized via an amide coupling reaction between an acetal-protected aminobenzaldehyde and 4-fluorobenzoic acid. After subsequent deprotection of the acetal to an aldehyde and reductive amination with tert-butyl (3-(methylamino)propyl)carbamate or tert-butyl 1,4-diazacycloheptane-1-carbamate, intermediates 36b and 41b were generated. Deprotection of the N-terminal Boc group and amide coupling yielded the final compounds 36 or 41. Conversely, compound 43 was synthesized in multiple steps, beginning with 1-(bromomethyl)-3-nitrobenzene and employing the Gabriel synthesis using the Ing-Manske procedure to generate the desired amine (precursor 42a). For ease of operation, a solution of 1,4-dioxane in 4M HCl was added to the product to precipitate the HCl salt of the amine. This precursor was then coupled with 5-phenylthiophene-2-carboxylic acid to give compound 42. Subsequently, the reduction of the nitro group and the subsequent amide coupling reaction yielded compound 43.

[0097] Precursor 46a of compounds 46 and 47 was synthesized in a two-step route. The mono-Boc protected piperazine was nucleophilically substituted with N-(3-bromopropyl)phthalimide to give intermediate 46b. Hydrazolysis in ethanol yielded the desired intermediate, which was used for amide coupling to give compound 46, or for reductive amination to give compound 47. Similarly, compound 48 was prepared by reductive amination of precursor 48a with phenylthiazolium carbaldehyde in dichloroethane solvent using sodium triacetoxyborohydride and triethylamine. This precursor was synthesized from compound 37e, in this case by coupling compound 37e with benzylamine (rather than with tert-butylamine in the synthesis of compound 37). Subsequent deprotection yielded the desired intermediate 48a.

[0098] Option 2. Compounds 33-46 a The synthesis of.

[0099]

[0100] a Reagents and conditions: (a) the corresponding amine, DIPEA, HATU, DMF, rt, 16 h, 9% – quantitative; (b) 1,4-dioxane solution of 4M HCl, rt, 1 h, quantitative; (c) benzaldehyde, NaBH(OAc)3, Et3N, DCE, rt, 16 h, 30%; (d) Pd / C 10%, H2, EtOH, rt, 16 h.

[0101] Scheme 3. Compounds 47-49 a The synthesis of.

[0102]

[0103] a Reagents and conditions: (a) the corresponding amine, NaBH(OAc)3, AcOH or Et3N, THF or DCE, rt, 16 h, 7%-71%; (b) amine, DIPEA, HATU, DMF, rt, 16 h, 46%.

[0104] Structure-activity relationship studies of initial seed compounds. Based on compounds from Yoshikawa et al. ( Figure 1 (EC) 50 Starting with a concentration of 2.21 µM, this compound was chosen to construct a more flexible core structure by removing the N-methylamide moiety from the thieno[3,2-c]quinoline “head”. This newly released phenylthiophene compound 1 (EC) 50= 0.416 µM) now shows a ~5-fold increase in potency in β-inhibitory protein recruitment assays. However, attempts to modify this moiety into different aromatic structures (6–7) depleted the compounds' β-inhibitory protein recruitment activity. Attempts to fuse the phenyl moiety with a five-membered heteroaromat (compounds 3–5) also reduced activity, where 3 (EC) 50 = 0.775 µM) is the best among this group of molecules. Therefore, our work focuses on the derivatization of the phenylthiophene moiety. Stereo-demanding substituents such as (tert-butyl)phenyl (10) and methoxyphenyl (11) reduce reactivity, while smaller substituents are generally tolerable. Of particular interest is the difluoro compound 17 (EC 50 = 0.758 µM) reduced activity, but some monofluorinated compounds (e.g., compound 16) increased activity. These molecules exhibited regio-specific preference. Most notably, meta-substitution (16, EC 0.758 µM) reduced activity. 50 =0.267 µM) significantly enhanced activity. A summary of our findings regarding "head" optimization, as described above, indicates that the phenylthiophene moiety is highly conserved in the binding of ACKR3. Only small substituents, such as fluorine, are tolerated. The enhanced binding affinity suggests that interactions with fluorine within the binding pocket, or the modulation of lipophilicity via fluorination, may play a crucial role.

[0105] The right side (“tail”) of the molecule is treated in a similar manner. Initially, the benzyl moiety is exchanged for the motif, which requires a larger space. This results in loss of agonistic activity, as seen in compound 36 (EC). 50 = 2.1 µM) and 37 (EC 50 =3.2 µM). Focusing on smaller modifications similar to “head” optimization, substituents such as para-methoxy (18), para-bromo (19), or para-fluorinated (20) were utilized. Again, fluorination proved to be the best choice, with the m-fluorobenzyl derivative (22) exhibiting the same potency as its meta-fluorinated “head” counterpart. Given that lipophilicity is an important factor, a dihydroisoquinoline derivative (35) with greater steric requirements but stronger lipophilicity was synthesized. Interestingly, this compound enhanced agonistic activity, becoming our most potent candidate to date (EC). 50 = 0.150 µM).

[0106] Next, the optimization process focused on methyl substitution of tertiary amines. Initially, compound 24 (EC24) with a larger space requirement was used. 50 = 0.709 µM) and 25 (EC 50The synthesis of ethylamine derivative 23 (EC = 0.630 µM) was used to challenge the spatial extent of the binding pocket. Although both compounds are still in the nanomolar range, they moderately reduce the activity of the initial seed compound. A more plausible approach is to examine the spatial extent of the binding pocket by extending the alkyl chain of the tertiary amine. 50 = 0.111 µM) greatly improved the potency and encouraged further elongation of the alkyl chain. Although propylamine derivative 26 (EC 50 = 0.111 µM) did not enhance activity, but butylamine derivative 27 (EC) 50 = 0.069 µM) showed excellent potency (in the lower nanomolar range). Therefore, the long nonpolar unsaturated alkyl chain further supports our hypothesis that lipophilicity may play a key role in the structure-activity relationship.

[0107] Conversely, rigidification of the "connector" portion eliminates the β-repressor protein recruitment efficacy of these compounds. For example, those with aromatic structural portions and those with 36 (EC) 50 Similar structures of 43 (EC = 2.1 µM) 50 = 3.35 µM) exhibits moderate to low agonistic activity. Non-aromatic cyclization pathways, such as diazacycloheptane derivative 40 (EC 10), also exhibit this activity. 50 = 1.2 µM) and 41 (EC 50 =76 µM), which also reduced the efficacy by at least 3 times.

[0108] Other experiments, such as changing the thiophene ring to a more polar thiazole (derivative "structure E", Table 1), resulted in the loss of agonistic activity. This derivatization of the compound's skeleton resulted in compound 48 having a maximum EC50 of 1.5 µM. 50 .

[0109] Shorter molecules were also synthesized, but were not sufficiently characterized further, such as 49 with a short linker chain and a fused benzothiophene core at 4.55 µM.

[0110] Efficacy of derivatives. In the β-inhibitor recruitment assay, the endogenous ligand CXCL12 of ACKR3 was set as 100% efficacy and used as a standard for determining the efficacy of individual compounds. As shown in Table 1, most of the tested compounds were found to be complete agonists of ACKR3. Lead compound 1 showed good efficacy of 71%, which was further improved with optimization work targeting β-inhibitor recruitment and platelet degranulation activity. Most changes were well tolerated and did not affect the efficacy of the compounds. Notably, highly potent compounds such as 16(E max = 103%), 23 (E max= 95%) or 26(E) max = 96%) showed complete agonist properties against ACKR3. At 82% efficacy, lead compound 27 was confirmed as a complete ACKR3 agonist, while also exhibiting excellent β-repressor protein recruitment efficacy in the low nanomolar concentration range.

[0111] Platelet degranulation studies of derivatives. While optimizing the β-inhibitor protein recruitment efficacy, the compounds were also tested and developed based on their functional activities. To determine the compounds' ability to regulate platelets, the surface expression of P-selectin was measured. P-selectin is a surface protein involved in cell adhesion mechanisms and can be found on activated platelets. Treatment with the corresponding compounds after inducing platelet aggregation in platelet-rich plasma (PRP) should therefore result in a significant reduction in P-selectin expression. P-selectin concentration was determined using an antibody conjugated with a fluorescent dye against P-selectin (compared to untreated PRP). The initial lead compound 1 (reduction in P-selectin expression = 71%) was demonstrated to be a potent inhibitor of platelet degranulation. Although structural changes in the “head” portion of 1 (structure A, Table 1) improved the β-inhibitor protein recruitment activity of the compound, the functional activity was preserved and, in some cases, even increased to near 100% reduction in P-selectin expression. In particular, compounds 16 and 31 exhibited high platelet activity, with efficacy up to 1.5 times higher than compound 1. Of the compounds tested from the "head" scaffold, only compound 5 was inactive in terms of functional activity. Furthermore, the compounds showed resistance to changes on the left side of the molecule. Conversely, changes in the "tail" compartment showed a major effect on platelet regulatory activity. Further rigidification (such as compound 35) led to decreased functional activity, while substitution of the benzyl moiety at the para position appeared to enhance the reduction in P-selectin expression. Considering β-inhibitor protein recruitment and flow cytometry assays, compound 20 (EC... 50 = 0.370 µM; P-selectin expression reduction = 86%) was the best molecule in this group. Interestingly, unlike para-fluorination of compound 20, meta- and ortho-fluorination decreased functional activity, with compound 22 showing a 66% reduction and compound 21 showing a 40% reduction. Similarly, platelet degranulation efficacy was highly sensitive to “connector” changes (structure C, Table 1). No obvious trend was identified. Using compounds 23, 26, and 27 as the most potent molecules in recruiting β-inhibitory proteins, they were also shown to be potent platelet regulators. Most compounds showed superior functional activity compared to the commercially available reference compound VUF11207 (P-selectin expression reduction = 65%).

[0112] In fact, compound 27 exhibits excellent β-repressor protein recruitment activity (EC). 50 = 69 nM) and high P-selectin expression decreased (77%), and was found to be the substance of most interest for further evaluation.

[0113] Table 1. Structure-activity relationship study.

[0114]

[0115]

[0116]

[0117]

[0118]

[0119] a The average of three independent experiments. b The efficacy of CXCL12 at 300 nM was set at 100%; the average of three independent experiments. c PRP was pre-incubated with the compound (100 µM) and then treated with CRP-XL (1 µg / mL). Data are presented as percentages (% ± SEM) of P-selectin-MFI normalized to CRP-XL-treated mediator PRP (DMSO 0.1%) from at least three independent experiments (in duplicate). A dash (–) = not tested.

[0120] Selectivity studies. To evaluate the selectivity of the selected most promising compounds, these compounds were tested against multiple receptors (ACKR2, CXCR3, and CXCR4). In particular, the selectivity of CXCR4 was expected, as its activation typically leads to physiological effects opposite to those of ACKR3.

[0121] All compounds exhibited high selectivity for the receptors described in Table 2. Figure 2 As shown, all the compounds tested had potency for these receptors that exceeded our measured limits and were all measured to be above 10 µM.

[0122] Table 2: Selective Studies

[0123]

[0124] a The average value (µM) of three independent experiments.

[0125] Cell viability studies. To further evaluate their toxicity, compounds 23 and 27 were tested in a cell viability assay. In HEK293 cells, both compounds showed superior quality in terms of cytotoxicity compared to VUF11207. Both tested compounds were well tolerated by HEK293 cells at concentrations below 1 µM. Even during longer test periods of up to 72 h, no effect on cells was observed during compound treatment. Figure 5 ).

[0126] Platelet aggregation studies. To further evaluate the inhibitory potential of these compounds on platelet aggregation, aggregation experiments were performed. Compared with the control, all tested compounds (including the reference compound VUF11207) showed a reduction in platelet aggregation. Notably, compound 23 showed the highest potency in inhibiting platelet aggregation. Combined with its excellent potency in FACS experiments, compound 23 demonstrates great potential as a tool compound, as... Figure 3 As shown.

[0127] Studies on metabolic stability. For example... Figure 4 As shown, compound 27 was stable in human liver microsomes over a period of 60 min. Approximately 69% of compound 27 remained unchanged, while the remainder was identified as a mixture of different metabolites. The most prominent metabolite was identified as the debenzylated compound, followed by the dealkylation of the N-butyl chain in terms of metabolite quantity. Subsequently, the metabolism of the compound was further studied using mouse liver microsomes. In this assay, the substance exhibited weak stability, with approximately 24% remaining after 60 min. In summary, although the compound has shown great promise in terms of human metabolic activity, further optimization is needed for its stability in mouse liver microsomes. Furthermore, in vitro metabolic stability studies of compounds 1, 12, 23, 29, and 30 in mouse liver microsomes are shown. Figure 6 ).

[0128] Solubility studies: The solubility of compounds 26 and 30 has been tested (Table 3). Both compounds showed good solubility in a mixture of 1 v / v DMSO and 99 v / v PBS buffer.

[0129] Table 3. Kinetic solubility study of the selected compounds.

[0130]

[0131] aThe average of three independent experiments (µM ± SEM). The compound was dissolved in 1% DMSO, mixed with 99% PBS buffer (pH 7.4), and then centrifuged at 16,000 g for 20 min. Solubility was determined by comparison with a standard curve of the corresponding compound.

[0132] It should be understood that, without departing from the scope of the invention, the features mentioned above and those to be mentioned below can be used not only in the combinations specified in the respective cases, but also in other combinations or separately.

[0133] The invention will now be further described with reference to embodiments that produce the additional features, characteristics, and advantages of the invention. These embodiments are merely illustrative and do not limit the scope or range of the invention. Features mentioned in a particular embodiment are general features of the invention and are applicable not only to that particular embodiment but also separately in the context of any other embodiment of the invention.

[0134] The invention will now be described and illustrated in further detail with reference to the following non-limiting embodiments and accompanying drawings. Attached Figure Description

[0135] Figure 1 An optimization process is shown, starting with a compound developed by Yoshikawa Y. et al.

[0136] Figure 2 The β-repressor protein recruitment activity of the selected most potent compound for CXCR4 is shown.

[0137] Figure 3 Platelet aggregation was shown after pre-incubation with 100 µM ACKR3 agonist and subsequent activation with 2.5 µM ADP.

[0138] Figure 4 The in vitro metabolic stability study of lead compound 27 in human liver microsomes is shown.

[0139] Figure 5 Cell viability assays of compounds 23 and 27 at 1 h, 24 h, 48 h, and 72 h compared to reference compound VUF11207 are shown.

[0140] Figure 6 The in vitro metabolic stability of compounds 1, 16, 26, 27 and 35 in mouse liver microsomes is shown.

[0141] Example

[0142] Chemistry and Methods.

[0143] Starting materials, reagents, and (anhydrous) solvents were commercially available and purchased from a range of manufacturers, and used without further purification. Thin-layer chromatography (TLC) with Macherey-Nagel pre-coated 60 F254 silica plates was used for reaction control. TLC spots were visualized by ultraviolet (UV) light (254 nm / 365 nm) or staining solutions. For purification purposes, rapid column chromatography was performed using a Grace Davison Davisil LC60A (20–45 μm) or Merck Geduran Si60 (mesh 63–200 μm) automated rapid chromatography system from LaFlash (VWR International GmbH, Darmstadt, Germany). NMR spectra were recorded at room temperature on a Bruker Avance 400 MHz spectrometer. Chemical shifts (δ) are reported in parts per million (ppm) relative to the internal standard tetramethylsilane (TMS), and the spectra are calibrated against residual solvent peaks for the deuterated solvents used (DMSO-d6 or CDCl3). Coupling constants (J) are expressed in Hertz (Hz). The purity of all compounds was determined by RP-HPLC using an Agilent 1100 series LC with a Phenomenex Luna C8 column (150 x 4.6 mm, 5 μm) and detected by a UV DAD detector at wavelengths of 254 nm and 230 nm. Elution was performed using the following gradient [A = 0.01 M KH2PO4, pH 2.30, B = MeOH] from 40% B to 85% B over 8 min, held at 85% B for 5 min, from 85% B to 40% B over 1 min, held at 40% B for 2 min, with a stop time of 16 min, at a flow rate of 1.5 mL / min. Mass spectrometry analyses were performed using an Advion Expression compact mass spectrometer (electrospray ionization, ESI) with a TLC plate readout system (using the following settings: ESI voltage 3.50 kV, capillary voltage 187 V, source voltage 44 V, capillary temperature 250 °C, desolvation gas temperature 250 °C, gas flow rate 5 L / min). High-resolution mass spectra (HRMS) of the final compounds were determined by the Mass Spectrometry Laboratory of the Institute of Organic Chemistry, Eberhard Karls University, Tübingen, on a Bruker maXis 4G ESI-TOF (Bruker Daltonik GmbH, Bremen, Germany).The instrument was operated in ESI positive ion mode with the following settings: nebulizer gas 1.2 bar, gas flow rate 6.0 L / min, source temperature 200 °C, capillary voltage +4500 V, and endplate bias voltage −500 V. The m / z range was from 80 to 1050 m / z. The purity of all final compounds was >95% as determined by the aforementioned HPLC procedure. NMR spectra of the compounds can be found on pages S26–S68 of SI. The synthesis of the precursors is available on pages S4–S25 of SI.

[0144] Synthesis. General Procedure I: Amide Coupling. The carboxylic acid was dissolved in anhydrous DMF (2.5 mL), and HATU (1.2 eq.) was slowly added to the solution. After stirring for 15 min, an amine (1 eq.) and DIPEA (3 eq.) were added to the mixture. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was diluted with DCM (10 mL) and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over anhydrous Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography or vacuum filtration.

[0145] General Procedure II: Suzuki Coupling. Add 1,4-dioxane (2 mL) and water (0.5 mL) to a flask containing an organohalide, boric acid or pinacol borate (1.1 eq.), XPhos Pd G4 (0.05 eq.), and K2CO3 (3 eq.). Degas the mixture with argon and heat to 100 °C for 16 h. After the reaction is complete, dilute the mixture with EtOAc (10 mL) and wash with water (3 x 10 mL) and brine (3 x 10 mL). Dry the combined organic phases with anhydrous Na2SO4, filter, and evaporate to dryness. Purify the crude product by rapid chromatography (SiO2; DCM / MeOH).

[0146] General Procedure III: Reductive Amination. The aldehyde (1.1 eq.) was dissolved in THF or DCE (5 mL), and one drop of acetic acid or triethylamine was added to the solution. After stirring for 15 min, the amine was added, and the mixture was stirred for 1 h. The mixture was cooled to 0 °C, and sodium triacetoxyborohydride (1.5 eq.) was added. After the reaction was complete, the mixture was diluted with EtOAc (10 mL) and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over anhydrous Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO₂; DCM / MeOH).

[0147] N-(3-(benzyl(methyl)amino)propyl)-5-phenylthiophene-2-carboxamide (1). Prepared according to general procedure 1 from 5-phenylthiophene-2-carboxylic acid (0.84 mmol) and N... 1 -Benzyl-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (1a, 0.84 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (2–7%). Yield: 150 mg (49%), yellow semi-solid. 1 ¹H NMR (400 MHz, CDCl₃) δ 8.02 (s, 1H), 7.59 (d, J = 7.2 Hz, 2H), 7.46–7.28 (m, 9H), 7.21 (d, J = 3.9 Hz, 1H), 3.74 (s, 2H), 3.56 (q, J = 11.4, 5.5 Hz, 2H), 2.80 (t, 2H), 2.42 (s, 3H), 1.92 (quintet, 2H). 13 C NMR (101 MHz, CDCl3) δ 162.84, 149.07, 137.46, 135.50, 133.70, 130.00, 129.48, 129.17,128.95, 128.61, 128.43, 126.22, 123.72, 62.68, 56.20, 40.74, 39.30, 25.16.TLC-MS (ESI+): C 20 H 24 The theoretical m / z value for N2OS is 364.16. The measured value is 365.2 [M+H]. + HPLC t R =5.778 min.

[0148] N-(3-(benzyl(methyl)amino)propyl)-5-bromothiophene-2-carboxamide (2). Prepared according to general procedure 1 from 5-bromothiophene-2-carboxylic acid (1.04 mmol) and N 1 -Benzyl-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (1a, 1.04 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (3–7%). Yield: 235 mg (53%), pale brown solid. 1¹H NMR (400 MHz, CDCl₃) δ 8.24 (s, 1H), 7.36 – 7.26 (m, 5H), 6.93 (dd, J = 12.9, 3.9 Hz, 2H), 3.54 (s, 2H), 3.50 (q, J = 10.4, 4.1 Hz, 2H), 2.61 (t, 2H), 2.27 (s, 3H), 1.78 (quintet, J = 11.6, 5.8 Hz, 2H). 13 C NMR (101 MHz, CDCl3) δ 160.85, 141.21, 138.02, 130.58, 129.62, 128.62, 127.84, 127.68,117.28, 63.37, 57.37, 50.97, 41.62, 40.66, 25.05. TLC-MS (ESI+): C 16 H 19 The theoretical m / z values ​​for BrN2OS are 366.04 / 369.04. The measured values ​​are 367.1 / 369.2 [M+H]. + HPLC t R = 4.709 min.

[0149] N-(3-(benzyl(methyl)amino)propyl)benzo[b]thiophene-2-carboxamide (3). Prepared from benzo[b]thiophene-2-carboxylic acid (0.6 mmol) and N according to general procedure 1. 1 -Benzyl-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (1a, 0.6 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (2–5%). Yield: 73 mg (36%), yellow solid. 1 ¹H NMR (400 MHz, CDCl₃) δ 8.41 (t, 1H), 7.82 (d, J = 7.6 Hz, 1H), 7.71 (d, J = 7.3 Hz, 1H), 7.60 (s, 1H), 7.44 – 7.33 (m, 4H), 7.34 – 7.27 (m, 3H), 3.70 (s, 2H), 3.58 (q, J = 10.9, 5.3 Hz, 2H), 2.77 (t, J = 5.7 Hz, 2H), 2.39 (s, 3H), 1.91 (quintet, 2H). 13C NMR (101 MHz, CDCl3) δ 162.7, 141.0, 139.4, 139.1,130.0, 128.8, 128.2, 126.2, 125.2, 125.1, 124.8, 122.7, 62.8, 56.5, 41.0,39.9,25.0. TLC-MS (ESI+): C 20 H 22 The theoretical m / z value for N2OS is 338.15. The measured value is 339.2 [M+H]. + HPLC t R = 4.067 min.

[0150] N-(3-(benzyl(methyl)amino)propyl)-1H-indole-2-carboxamide (4). Prepared according to general procedure 1 from 1H-indole-2-carboxylic acid (0.84 mmol) and N... 1 -Benzyl-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (1a, 0.84 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (2–7%). Yield: 151 mg (56%), yellow solid. 1 HNMR (400 MHz, CDCl3) δ 9.73 (s, 1H), 8.20 (t, 1H), 7.47 (d, J = 8.0 Hz, 1H), 7.37 (d, J = 8.2 Hz, 1H), 7.32 – 7.22 (m, J = 14.0, 6.8, 3.5 Hz, 5H), 7.18(td, J = 15.3, 0.9 Hz, 1H), 7.04 (t, J = 7.5 Hz, 1H), 6.55 (s, 1H), 3.56 (s,2H), 3.52 (q, J = 11.3, 5.5 Hz, 2H), 2.73 – 2.55 (m, 2H), 2.26 (s, 3H), 1.78 (quintet, 2H). 13 C NMR (101 MHz, CDCl3) δ 162.14, 136.90, 136.49, 131.06,129.86, 128.82, 128.03, 127.80, 124.34, 121.99, 120.54, 112.18, 102.62,63.03, 56.85, 41.10, 39.65, 25.25. TLC-MS (ESI+): C 20 H23 The theoretical m / z value for N3O is 318.18. The measured value is 322.2 [M+H]. + HPLC t R = 1.089 min.

[0151] N-(3-(benzyl(methyl)amino)propyl)benzofuran-2-carboxamide (5). Prepared from benzofuran-2-carboxylic acid (0.16 mmol) and N according to general procedure 1. 1 -Benzyl-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (1a, 0.16 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (0–4%). Yield: 15 mg (30%), yellow semi-solid. 1 HNMR (400 MHz, CDCl3) δ 8.23 ​​(s, 1H), 7.62 (d, J = 7.8 Hz, 1H), 7.44 – 7.15 (m, 9H), 3.61 (s, 2H), 3.56 (q, J = 11.2, 5.5 Hz, 2H), 2.64 (t, J = 5.9 Hz, 2H), 2.30 (s, 3H), 1.85 (quintet, 2H). 13 C NMR (101 MHz, CDCl3) δ 159.2, 154.9,149.3, 129.6, 128.7, 127.8, 127.7, 126.7, 123.7, 122.7, 111.9, 110.0, 63.0,56.4, 41.6, 39.2, 29.8, 25.7. TLC-MS (ESI+): C 20 H 22 The theoretical m / z value for N₂O₂ is 322.17. The measured value is 323.2 [M+H]. + HPLC t R = 4.123 min.

[0152] N-(3-(benzyl(methyl)amino)propyl)-4-fluorobenzamide (6). Prepared according to general procedure 1 from 4-fluorobenzoic acid (0.33 mmol) and N 1 -Benzyl-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (1a, 0.33 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (3–7%). Yield: 41 mg (41%), brown semi-solid. 1¹H NMR (400MHz, CDCl₃) δ 8.17 (s, 1H), 7.72 – 7.60 (m, J = 8.4, 5.5 Hz, 2H), 7.31 – 7.18 (m, J = 1.8 Hz, 5H), 6.99 (t, J = 8.6 Hz, 2H), 3.63 – 3.46 (m, 4H), 2.64 (t, 2H), 2.26 (s, 3H), 1.82 (quintet, J = 11.4, 5.8 Hz, 2H). 13 C NMR (101 MHz, CDCl3) δ 166.29, 165.86, 163.37, 137.98, 130.90, 130.87, 129.53, 129.38,129.29, 128.56, 127.60, 115.53, 115.32, 63.22, 57.21, 41.53, 40.45, 25.25. TLC-MS (ESI+): C 18 H 21 The theoretical m / z value for FN₂O is 300.16. The measured value is 301.2 [M+H]. + HPLC t R =3.618 min.

[0153] N-(3-(benzyl(methyl)amino)propyl)-2-(phenylthio)acetamide (7). Prepared according to general procedure 1 from 2-(phenylthio)acetic acid (0.36 mmol) and N... 1 -Benzyl-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (1a, 0.36 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (1–4%). Yield: 28 mg (24%), yellow oil. 1 HNMR (400 MHz, CDCl3) δ 7.71 (s, 1H), 7.32 – 7.13 (m, 10H), 3.59 (s, 2H), 3.41 (s, 2H), 3.30 (q, J = 12.1, 5.9 Hz, 2H), 2.32 (t, J = 6.3 Hz, 2H), 2.09 (s, 3H), 1.60 (quintet, 2H). 13C NMR (101 MHz, CDCl3) δ 167.87, 138.30, 135.31,129.32, 129.31, 128.43, 127.57, 127.34, 126.36, 62.85, 55.35, 41.93, 39.11,37.12,26.03. TLC-MS (ESI+): C 19 H 24 The theoretical m / z value for N2OS is 328.16. The measured value is 329.2 [M+H]. + HPLC t R = 4.079 min.

[0154] N-(3-(benzyl(methyl)amino)propyl)-3-(2-fluorophenyl)propionamide (8). Prepared according to general procedure 1 from 3-(2-fluorophenyl)propionic acid (1.67 mmol) and N... 1 -Benzyl-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (1a, 1.67 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (2–6%). Yield: 470 mg (86%), brown semi-solid. 1 ¹H NMR (400 MHz, CDCl₃) δ 7.32 – 7.19 (m, 5H), 7.19 – 7.09 (m, 2H), 7.04 – 6.89 (m, 2H), 6.72 (t, 1H), 3.47 (s, 2H), 3.25 (q, 2H), 2.91 (t, J = 7.7 Hz, 2H), 2.42 (t, J = 6.1 Hz, 2H), 2.36 (t, 2H), 2.17 (s, 3H), 1.61 (quintet, 2H). 13 C NMR (101 MHz, CDCl3) δ 172.28, 162.39, 159.96, 137.72, 130.87,130.82, 129.31, 128.61, 128.10, 128.02, 127.87, 127.71, 127.66, 124.19,124.15, 115.40, 115.18, 62.61, 55.41, 41.51, 38.57, 36.80, 25.68, 25.33,25.31. TLC-MS (ESI+): C 20 H 25The theoretical m / z value for FN₂O is 328.20. The measured value is 329. [M+H] + HPLC t R =4.507 min.

[0155] N-(3-(benzyl(methyl)amino)propyl)-5-(p-tolyl)thiophene-2-carboxamide (9). Prepared from N-(3-(benzyl(methyl)amino)propyl)-5-bromothiophene-2-carboxamide (2, 0.22 mmol) and p-tolylboronic acid (0.24 mmol, 1.1 eq.) according to general procedure 2. Purified by rapid chromatography using DCM / MeOH (4–8%). Yield: 30 mg (33%), brown semi-solid. 1 ¹H NMR (400 MHz, CDCl₃) δ 8.06 (s, 1H), 7.35 (d, J = 7.2 Hz, 1H), 7.29 – 7.20 (m, 9H), 6.92 (d, J = 3.8 Hz, 1H), 3.56 – 3.48 (m, 4H), 2.58 (t, 2H), 2.39 (s, 3H), 2.27 (s, 3H), 1.78 (quintet, 2H). 13 C NMR (101 MHz, CDCl3) δ 161.93, 147.40, 138.91, 138.25, 136.17, 133.59, 130.98, 130.42,129.56, 128.53, 128.51, 128.02, 127.49, 126.91, 126.14, 63.30, 57.04, 41.82, 40.43, 25.43, 21.19. TLC-MS (ESI+): C 23 H 26 The theoretical m / z value for N2OS is 378.18. The measured value is 379.2 [M+H]. + HPLC t R = 6.242 min.

[0156] N-(3-(benzyl(methyl)amino)propyl)-5-(4-(tert-butyl)phenyl)thiophene-2-carboxamide (10). Prepared from N-(3-(benzyl(methyl)amino)propyl)-5-bromothiophene-2-carboxamide (2, 0.22 mmol) and (4-(tert-butyl)phenyl)boronic acid (0.25 mmol, 1.1 eq.) according to general procedure 2. Purified by rapid chromatography using DCM / MeOH (6–10%). Yield: 27 mg (26%), brown solid.1 ¹H NMR (400 MHz, CDCl₃) δ 8.06 (t, 1H), 7.43 (dd, J = 45.9, 8.4 Hz, 4H), 7.28 – 7.20 (m, 6H), 7.11 (d, J = 3.8 Hz, 1H), 3.54 – 3.46 (m, 4H), 2.57 (t, J = 5.8 Hz, 2H), 2.25 (s, 3H), 1.76 (quintet, 2H), 1.30 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 161.94, 151.72, 148.49, 138.17, 137.85,131.05, 129.62, 128.86, 128.57, 127.53, 126.04, 125.91, 122.99, 63.30, 57.10,41.74, 40.45, 34.80, 31.35, 25.38. TLC-MS (ESI+): C 26 H 32 The theoretical m / z value for N2OS is 420.22. The measured value is 421.2 [M+H]. + HPLC t R = 7.967 min.

[0157] N-(3-(benzyl(methyl)amino)propyl)-5-(4-methoxyphenyl)thiophene-2-carboxamide (11). Prepared from N-(3-(benzyl(methyl)amino)propyl)-5-bromothiophene-2-carboxamide (2, 0.19 mmol) and (4-methoxyphenyl)boronic acid (0.21 mmol, 1.1 eq.) according to general procedure 2. Purified by rapid chromatography using DCM / MeOH (4–8%). Yield: 57 mg (71%), yellowish-brown solid. 1 ¹H NMR (400 MHz, CDCl₃) δ 7.95 (t, 1H), 7.50 – 7.42 (m, 2H), 7.30 – 7.16 (m, J = 12.8, 7.9, 4.7 Hz, 6H), 7.01 (d, J = 3.8 Hz, 1H), 6.90 – 6.82 (m, 2H), 3.78 (s, 3H), 3.53 – 3.45 (m, 4H), 2.55 (t, 2H), 2.23 (s, 3H), 1.74 (quintet, 2H). 13C NMR (101 MHz, CDCl3) δ 161.92, 159.98,148.39, 138.33, 137.33, 129.57, 128.90, 128.53, 127.46, 126.65, 122.33,114.51, 63.32, 57.14, 55.48, 41.76, 40.40, 25.50. TLC-MS (ESI+): C 23 H 26 The theoretical m / z value for N₂O₂S is 394.17. The measured value is 395. [M+H] + HPLC t R = 5.547 min.

[0158] N-(3-(benzyl(methyl)amino)propyl)-5-(4-hydroxyphenyl)thiophene-2-carboxamide (12). Prepared from N-(3-(benzyl(methyl)amino)propyl)-5-bromothiophene-2-carboxamide (2, 0.21 mmol) and (4-hydroxyphenyl)boronic acid (0.23 mmol, 1.1 eq.) according to general procedure 2. Purified by rapid chromatography using DCM / MeOH (5–8%). Yield: 21 mg (24%), brown solid. 1 H NMR (400 MHz, CDCl3) δ 8.14 (s, 1H), 7.37 (d, J = 10.8Hz, 2H), 7.28 – 7.17 (m, J = 7.6, 3.5 Hz, 6H), 6.99 (d, J = 3.8 Hz, 1H), 6.87(d, J = 8.4 Hz, 2H), 6.36 (s, 1H), 3.56 (s, 2H), 3.51 (q, J = 4.5 Hz, 2H), 2.61 (t, J = 5.8 Hz, 2H), 2.27 (s, 3H), 1.80 (quint, 2H). 13 C NMR (101 MHz, CDCl3) δ 162.56, 157.58, 149.40, 137.31, 136.10, 129.83, 129.49, 128.65,127.78, 127.63, 125.63, 122.15, 116.30, 63.03, 56.80, 41.50, 40.39, 25.22. TLC-MS (ESI+): C 22 H 24The theoretical m / z value for N₂O₂S is 380.16. The measured value is 381.2 [M+H]. + HPLC t R =4.041 min.

[0159] N-(3-(benzyl(methyl)amino)propyl)-5-(pyridin-4-yl)thiophene-2-carboxamide (13). Prepared from N-(3-(benzyl(methyl)amino)propyl)-5-bromothiophene-2-carboxamide (2, 0.27 mmol) and pyridin-4-ylboronic acid hydrate (0.3 mmol, 1.1 eq.) according to general procedure 2. Purified by rapid chromatography using DCM / MeOH (5–8%). Yield: 36 mg (33%), yellow oil. 1 ¹H NMR (400 MHz, CDCl₃) δ 8.58 (d, J = 5.6 Hz, 2H), 8.35 (s, 1H), 7.40 (d, J = 4.7, 1.5 Hz, 2H), 7.30 (d, J = 4.5 Hz, 1H), 7.28 –7.23 (m, 5H), 7.21 (d, J = 3.9 Hz, 1H), 3.57 – 3.49 (m, 4H), 2.61 (t, 2H), 2.25 (s, 3H), 1.78 (quintet, 2H). 13 C NMR (101 MHz, CDCl3) δ 161.25, 150.62,144.61, 140.95, 140.83, 138.18, 129.67, 128.71, 128.58, 127.59, 125.52,120.06, 63.41, 57.56, 41.66, 40.82, 25.11. TLC-MS (ESI+): C 21 H 23 The theoretical m / z value for N3OS is 365.16. The measured value is 366.2 [M+H]. + HPLC t R = 1.643 min.

[0160] N-(3-(benzyl(methyl)amino)propyl)-5-(4-fluorophenyl)thiophene-2-carboxamide (14). Prepared from N-(3-(benzyl(methyl)amino)propyl)-5-bromothiophene-2-carboxamide (2, 0.26 mmol) and 2-(4-fluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxoboronylcyclopentane (0.29 mmol, 1.1 eq.) according to general procedure 2. Purified by rapid chromatography using DCM / MeOH (2–6%). Yield: 20 mg (18%), brown semi-solid. 1 H NMR (400 MHz, CDCl3) δ8.16 (s, 1H), 7.59 – 7.49 (m, J = 8.1, 5.1, 2.5 Hz, 2H), 7.33 – 7.27 (m, J =8.1, 3.9 Hz, 5H), 7.25 – 7.22 (m, J = 3.7 Hz, 1H), 7.14 – 7.02 (m, 3H), 3.59 – 3.51 (m, 4H), 2.67 – 2.58 (m, 2H), 2.29 (s, 3H), 1.86 – 1.76 (m, 2H). 13 C NMR(101 MHz, CDCl3) δ 164.14, 161.75, 147.19, 138.49, 138.08, 130.15, 130.12,129.69, 128.83, 128.59, 127.96, 127.88, 127.59, 123.38, 116.23, 116.02, 63.33, 57.27, 41.65, 40.54, 25.29. TLC-MS (ESI+): C 22 H 23 The theoretical m / z value for FN2OS is 382.15. The measured value is 383.2 [M+H]. + HPLC t R = 5.957 min.

[0161] N-(3-(benzyl(methyl)amino)propyl)-5-(2-fluorophenyl)thiophene-2-carboxamide (15). Prepared from N-(3-(benzyl(methyl)amino)propyl)-5-bromothiophene-2-carboxamide (2, 0.22 mmol) and (2-fluorophenyl)boronic acid (0.24 mmol, 1.1 eq.) according to general procedure 2. Purified by rapid chromatography using DCM / MeOH (2–6%). Yield: 20 mg (22%), brown semi-solid. 1¹H NMR (400 MHz, CDCl₃) δ 8.14 (s, 1H), 7.54 (t, 1H), 7.33 – 7.29 (m, J = 3.0 Hz, 1H), 7.29 – 7.20 (m, 7H), 7.17 – 7.06 (m, J = 16.5, 8.6Hz, 2H), 3.59 – 3.44 (m, 4H), 2.58 (t, 2H), 2.26 (s, 3H), 1.77 (quintet, 2H). 13 C NMR (101 MHz, CDCl3) δ 161.79, 160.58, 158.09, 141.14, 141.11, 139.17,139.14, 137.96, 129.70, 129.66, 129.61, 128.98, 128.95, 128.59, 128.47,127.60, 126.78, 126.71, 124.71, 124.67, 121.82, 121.69, 116.68, 116.46,63.24, 56.98, 41.69, 40.45, 25.28. TLC-MS (ESI+): C 22 H 23 The theoretical m / z value for FN2OS is 382.15. The measured value is 383.2 [M+H]. + HPLC t R = 6.044 min.

[0162] N-(3-(benzyl(methyl)amino)propyl)-5-(3-fluorophenyl)thiophene-2-carboxamide (16). Prepared from N-(3-(benzyl(methyl)amino)propyl)-5-bromothiophene-2-carboxamide (2, 0.22 mmol) and (3-fluorophenyl)boronic acid (0.24 mmol, 1.1 eq.) according to general procedure 2. Purified by rapid chromatography using DCM / MeOH (4–8%). Yield: 63 mg (72%), brown semi-solid. 1 ¹H NMR (400 MHz, CDCl₃) δ 8.13 (s, 1H), 7.35 – 7.22 (m, 8H), 7.21 (d, J = 3.9 Hz, 1H), 7.13 (d, J = 3.9 Hz, 1H), 7.02 – 6.93 (m, 1H), 3.56– 3.46 (m, 4H), 2.58 (t, 2H), 2.24 (s, 3H), 1.76 (quintet, 2H).13 C NMR (101MHz, CDCl3) δ 163.14, 161.57, 146.66, 139.17, 138.34, 135.95, 130.97, 130.71,129.60, 129.60, 128.77, 128.55, 128.55, 127.54, 124.06, 121.85, 115.07,113.09, 63.40, 57.39, 41.73, 40.63, 25.36. TLC-MS (ESI+): C 22 H 23 The theoretical m / z value for FN2OS is 382.15. The measured value is 383.2 [M+H]. + HPLC t R = 5.940 min.

[0163] N-(3-(benzyl(methyl)amino)propyl)-5-(3,5-difluorophenyl)thiophene-2-carboxamide (17). Prepared from N-(3-(benzyl(methyl)amino)propyl)-5-bromothiophene-2-carboxamide (2, 0.19 mmol) and (3,5-difluorophenyl)boronic acid (0.21 mmol, 1.1 eq.) according to general procedure 2. Purified by rapid chromatography using DCM / MeOH (2–7%). Yield: 26 mg (31%), brown semi-solid. 1 ¹H NMR (400 MHz, CDCl₃) δ 8.31 (s, 1H), 7.33 – 7.26 (m, 5H), 7.18 (dd, J = 18.3, 3.9 Hz, 2H), 7.10 – 7.01 (m, 2H), 6.82 – 6.72 (m, 1H), 3.59 – 3.50 (m, 4H), 2.66 – 2.61 (m, 2H), 2.28 (s, 3H), 1.80 (quintet, J = 11.5, 5.8 Hz, 2H). 13 C NMR (101 MHz, CDCl3) δ 163.36, 161.39, 145.39,139.79, 138.28, 136.81, 129.71, 128.82, 128.60, 127.65, 124.74, 109.05,103.57, 63.49, 57.71, 41.66, 40.92, 25.12. TLC-MS (ESI+): C 22 H 22The theoretical m / z value for F2N2OS is 400.14. The measured value is 401.2 [M+H]. + HPLC t R = 6.630 min.

[0164] N-(3-((4-methoxybenzyl)(methyl)amino)propyl)-5-phenylthiophene-2-carboxamide (18). Prepared according to general procedure 1 from 5-phenylthiophene-2-carboxylic acid (0.29 mmol) and N 1 -(4-Methoxybenzyl)-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (18a, 0.29 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (1–4%). Yield: 33 mg (28%), brown semi-solid. 1 H NMR (400 MHz, CDCl3) δ 8.20 (s, 1H), 7.61 – 7.55 (m, J = 7.1, 4.8, 2.6 Hz, 2H), 7.42 – 7.36 (m, 2H), 7.35 – 7.28 (m, J = 11.8, 6.0, 2.6 Hz, 2H), 7.25 – 7.21 (m, J = 8.6 Hz, 2H), 7.19 (d, J = 3.9 Hz, 1H), 6.87 – 6.79 (m, 2H), 3.74 (s, 3H), 3.59 – 3.49 (m, 4H), 2.64 (t, 2H), 2.31(s, 3H), 1.83 (quintet, J = 11.4, 5.8 Hz, 2H). 13 C NMR (101 MHz, CDCl3) δ161.93, 159.23, 148.35, 133.83, 131.25, 131.00, 129.23, 129.15, 129.05,128.47, 126.16, 123.45, 113.97, 62.51, 55.34, 29.85, 25.11, 22.84, 14.27. TLC-MS (ESI+): C 23 H 26 The theoretical m / z value for N₂O₂S is 394.17. The measured value is 395.2 [M+H]. + HPLC t R = 6.072min.

[0165] N-(3-((4-bromobenzyl)(methyl)amino)propyl)-5-phenylthiophene-2-carboxamide (19). Prepared according to general procedure 1 from 5-phenylthiophene-2-carboxylic acid (0.29 mmol) and N 1 -(4-Bromobenzyl)-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (19a, 0.29 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (1–4%). Yield: 63 mg (48%), yellow semi-solid. 1 ¹H NMR (400 MHz, CDCl₃) δ 7.92 (t, J = 4.6 Hz, 1H), 7.59–7.51 (m, J = 5.2, 3.3 Hz, 2H), 7.43–7.33 (m, 4H), 7.32–7.25 (m, 2H), 7.20–7.13 (m, J = 5.9, 5.1 Hz, 3H), 3.57–3.42 (m, J = 11.7, 5.7 Hz, 4H), 2.58 (t, J = 5.9 Hz, 2H), 2.25 (s, 3H), 1.78 (quintet, 2H). 13 C NMR (101 MHz, CDCl3)δ 161.99, 148.51, 137.87, 136.94, 133.60, 131.65, 131.23, 129.15, 129.08,128.49, 126.09, 123.48, 121.48, 62.39, 56.84, 41.48, 40.05, 25.42. TLC-MS (ESI+): C 22 H 23 The theoretical m / z value for BrN₂OS is 442.07. The measured value is 465.1 [M+Na]. + HPLC t R = 6.856 min.

[0166] N-(3-((4-fluorobenzyl)(methyl)amino)propyl)-5-phenylthiophene-2-carboxamide (20). Prepared according to general procedure 1 from 5-phenylthiophene-2-carboxylic acid (0.44 mmol) and N 1 -(4-Fluorobenzyl)-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (20a, 0.44 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (1–6%). Yield: 90 mg (53%), brown semi-solid.1 ¹H NMR (400 MHz, CDCl₃) δ 8.05 (s, 1H), 7.54 (d, J = 9.6 Hz, 2H), 7.40 – 7.33 (m, 2H), 7.33 – 7.29 (m, 2H), 7.28 – 7.23 (m, J = 8.1, 5.3Hz, 2H), 7.17 (d, J = 3.8 Hz, 1H), 6.96 (t, J = 8.6 Hz, 2H), 3.54 – 3.48 (m, 4H), 2.62 (t, J = 18.0, 12.1 Hz, 2H), 2.26 (s, 3H), 1.79 (quintet, 2H). 13 C NMR(101 MHz, CDCl3) δ 162.22, 162.05, 148.54, 137.89, 133.59, 133.45, 131.19,129.10, 129.07, 128.50, 126.07, 123.42, 115.38, 62.21, 56.77, 41.32, 40.10,25.35. TLC-MS (ESI+): C 20 H 23 The theoretical m / z value for FN2OS is 382.15. The measured value is 383.2 [M+H]. + HPLC t R = 6.094 min.

[0167] N-(3-((2-fluorobenzyl)(methyl)amino)propyl)-5-phenylthiophene-2-carboxamide (21). Prepared according to general procedure 1 from 5-phenylthiophene-2-carboxylic acid (0.29 mmol) and N 1 -(2-Fluorobenzyl)-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (21a, 0.29 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (1–4%). Yield: 100 mg (89%), yellow semi-solid. 1¹H NMR (400 MHz, CDCl₃) δ 8.16 (t, J = 4.7 Hz, 1H), 7.57 – 7.51 (m, 2H), 7.37 – 7.22 (m, 6H), 7.15 (d, J = 3.9 Hz, 1H), 7.10 – 7.04 (m, 1H), 7.04 – 6.97 (m, J = 9.5, 8.2, 1.0 Hz, 1H), 3.66 (s, 2H), 3.52 (q, 2H), 2.68 (t, J = 13.8, 7.9 Hz, 2H), 2.31 (s, 3H), 1.81 (quintet, 2H). 13 C NMR (101MHz, CDCl3) δ 162.88, 162.11, 160.42, 148.41, 138.14, 133.73, 132.24, 132.20,129.82, 129.73, 129.07, 128.93, 128.42, 126.09, 124.24, 124.20, 123.43,115.80, 115.58, 57.02, 55.69, 53.55, 41.24, 40.17, 25.23. TLC-MS (ESI+): C 22 H 23 The theoretical m / z value for FN2OS is 382.15. The measured value is 383.2 [M+H]. + HPLC t R = 5.901 min.

[0168] N-(3-((3-fluorobenzyl)(methyl)amino)propyl)-5-phenylthiophene-2-carboxamide (22). Prepared according to general procedure 1 from 5-phenylthiophene-2-carboxylic acid (0.29 mmol) and N 1 -(3-Fluorobenzyl)-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (22a, 0.29 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (1–4%). Yield: 33 mg (29%), yellow semi-solid. 1¹H NMR (400 MHz, CDCl₃) δ 7.88 (t, J = 4.4 Hz, 1H), 7.59 – 7.52 (m, 2H), 7.41 – 7.34 (m, 2H), 7.33 – 7.28 (m, 2H), 7.25 – 7.16 (m, 2H), 7.11 – 7.00 (m, 2H), 6.98 – 6.90 (m, 1H), 3.60 – 3.49 (m, 4H), 2.60 (t, 2H), 2.27 (s, 3H), 1.80 (quintet, 2H). 13 C NMR (101 MHz, CDCl3) δ 164.19, 161.97,161.74, 148.49, 140.71, 138.04, 133.72, 130.13, 130.05, 129.11, 128.96,128.49, 126.15, 125.11, 125.08, 123.48, 116.40, 116.19, 114.59, 114.38,62.69, 57.05, 41.64, 40.17, 25.52. TLC-MS (ESI+): C 22 H 23 The theoretical m / z value for FN2OS is 382.15. The measured value is 383.2 [M+H]. + HPLC t R = 6.155 min.

[0169] N-(3-(benzyl(ethyl)amino)propyl)-5-phenylthiophene-2-carboxamide (23). Prepared according to general procedure 1 from 5-phenylthiophene-2-carboxylic acid (0.44 mmol) and N 1 -Benzyl-N 1 1,3-Ethylpropane-1,3-diamine dihydrochloride (23a, 0.44 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (1–6%). Yield: 70 mg (42%), brown semi-solid. 1¹H NMR (400 MHz, CDCl₃) δ 7.94 (s, 1H), 7.61 – 7.52 (m, 2H), 7.39 – 7.33 (m, J = 10.1, 4.7 Hz, 2H), 7.32 – 7.26 (m, J = 8.3, 5.6, 3.3 Hz, 4H), 7.26 – 7.21 (m, 3H), 7.16 (d, J = 3.9 Hz, 1H), 3.58 (s, 2H), 3.47 (q, J = 7.8, 3.9 Hz, 2H), 2.66 – 2.52 (m, 4H), 1.75 (quintet, 2H), 1.08 (t, J = 7.1 Hz, 3H). 13 CNMR (101 MHz, CDCl3) δ 161.90, 148.32, 138.52, 138.28, 133.76, 129.53,129.08, 128.80, 128.51, 128.41, 127.38, 126.09, 123.33, 58.43, 52.45, 47.05, 40.34, 25.23, 11.11. TLC-MS (ESI+): C 23 H 26 The theoretical m / z value for N2OS is 378.18. The measured value is 379.2 [M+H]. + HPLC t R = 6.043 min.

[0170] N-(3-(benzyl(isopropyl)amino)propyl)-5-phenylthiophene-2-carboxamide (24). Prepared according to general procedure 1 from 5-phenylthiophene-2-carboxylic acid (0.42 mmol) and N 1 -Benzyl-N 1 -Isopropylpropane-1,3-diamine dihydrochloride (24a, 0.42 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (0–3%). Yield: 30 mg (18%), yellow semi-solid. 1¹H NMR (400 MHz, CDCl₃) δ 7.65 – 7.56 (m, 2H), 7.45 – 7.32 (m, 6H), 7.31 – 7.22 (m, J = 10.2, 6.9, 2.7 Hz, 3H), 7.19 – 7.12 (m, J = 15.7, 3.8 Hz, 2H), 3.56 (s, 2H), 3.51 – 3.44 (m, J = 11.6, 5.4 Hz, 2H), 3.15 – 3.01 (m, 1H), 2.60 (t, 2H), 1.72 (quintet, J = 11.6, 5.9 Hz, 2H), 1.07 (d, J = 6.6 Hz, 6H). 13 C NMR (101 MHz, CDCl3) δ 161.85, 148.36, 140.19, 138.31, 133.83, 129.39,129.12, 128.74, 128.54, 128.44, 127.22, 126.15, 123.28, 54.26, 49.00, 47.99, 39.94, 26.08, 17.52. TLC-MS (ESI+): C 24 H 28 The theoretical m / z value for N2OS is 392.19. The measured value is 393.2 [M+H]. + HPLC t R = 5.511 min.

[0171] N-(3-(benzyl(cyclopropyl)amino)propyl)-5-phenylthiophene-2-carboxamide (25). Prepared according to general procedure 1 from 5-phenylthiophene-2-carboxylic acid (0.54 mmol) and N 1 -Benzyl-N 1 1,3-Cyclopropylpropane-1,3-diamine (25a, 0.54 mmol, 1 eq.). Purified by rapid chromatography using PE / EtOAc (10–40%). Yield: 55 mg (26%), yellow semi-solid. 1H NMR (400 MHz, CDCl3) δ 7.65 – 7.55 (m, J = 5.2, 3.4 Hz, 2H), 7.42 –7.35 (m, 2H), 7.34 – 7.29 (m, 1H), 7.28 – 7.19 (m, 8H), 3.76 (s, 2H), 3.44(q, J = 11.6, 5.7 Hz, 2H), 2.71 (t, 2H), 1.86 – 1.72 (m, 3H), 0.57 – 0.51 (m,2H), 0.50 – 0.44 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 161.76, 148.35, 138.11,137.85, 133.79, 129.88, 129.16, 128.85, 128.50, 128.33, 127.34, 126.17,123.40, 60.04, 54.37, 40.31, 37.65, 25.37, 7.37. TLC-MS (ESI+): C 24 H 26 The theoretical m / z value for N2OS is 390.18. The measured value is 391.2 [M+H]. + HPLC t R = 6.192 min.

[0172] N-(3-(benzyl(propyl)amino)propyl)-5-phenylthiophene-2-carboxamide (26). Prepared according to general procedure 1 from 5-phenylthiophene-2-carboxylic acid (0.36 mmol) and N 1 -Benzyl-N 1 1,3-Propylpropane-1,3-diamine dihydrochloride (26a, 0.36 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (1–4%). Yield: 36 mg (26%), colorless oil. 1 ¹H NMR (400 MHz, CDCl₃) δ 7.66 – 7.56 (m, 3H), 7.46 – 7.30 (m, 8H), 7.22 (d, J = 3.9 Hz, 1H), 3.88 (s, 2H), 3.51 (q, 2H), 2.87 (s, 2H), 2.80 (s, 1H), 2.70 (t, 2H), 1.95 (quintet, 2H), 1.71 – 1.56 (m, 2H), 0.90 (t, J = 7.3 Hz, 3H).13 C NMR (101 MHz, CDCl3) δ 163.47, 149.50, 136.79, 133.57, 129.99, 129.73,129.18, 129.07, 128.68, 126.18, 123.86, 58.37, 54.95, 51.91, 38.72, 25.20,18.62, 11.56. TLC-MS (ESI+): C 24 H 28 The theoretical m / z value for N2OS is 392.19. The measured value is 393.2 [M+H]. + HPLC t R = 6.351 min.

[0173] N-(3-(benzyl(butyl)amino)propyl)-5-phenylthiophene-2-carboxamide (27). Prepared according to general procedure 1 from 5-phenylthiophene-2-carboxylic acid (0.46 mmol) and N 1 -Benzyl-N 1 1,3-Butylpropane-1,3-diamine dihydrochloride (27a, 0.46 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (0–3%). Yield: 13 mg (7%), yellow solid. 1 ¹H NMR (400 MHz, CDCl₃) δ 7.66 – 7.53 (m, J = 5.3, 3.3 Hz, 3H), 7.43 – 7.37 (m, 2H), 7.36 – 7.22 (m, 7H), 7.20 (d, J = 3.8 Hz, 1H), 3.60 (s, 2H), 3.50 (q, J = 11.5, 5.5 Hz, 2H), 2.60 (t, 2H), 2.51 (t, 2H), 1.77 (quintet, J = 11.4, 5.8 Hz, 2H), 1.60 – 1.48 (m, 2H), 1.35 – 1.26 (m, J = 15.0, 7.4 Hz, 2H). 0.88 (t, J = 7.3 Hz, 3H). 13C NMR (101 MHz, CDCl3) δ 161.91, 148.39,138.33, 133.86, 129.57, 129.15, 128.81, 128.57, 128.47, 127.43, 126.18,123.35, 59.08, 53.64, 52.94, 40.09, 28.73, 25.55, 20.87, 14.18. TLC-MS (ESI+): C 25 H 30 The theoretical m / z value for N2OS is 406.21. The measured value is 407.2 [M+H]. + HPLC t R = 6.820 min.

[0174] N-(3-(benzyl(3-hydroxypropyl)amino)propyl)-5-phenylthiophene-2-carboxamide (28). Prepared from 5-phenylthiophene-2-carboxylic acid (1.2 mmol) and 3-((3-aminopropyl)(benzyl)amino)prop-1-ol dihydrochloride (28a, 1.2 mmol, 1 eq.) according to general procedure 1. Purified by rapid chromatography using DCM / MeOH (0–4%). Yield: 25 mg (5%), colorless oil. 1 ¹H NMR (400 MHz, CDCl₃) δ 7.62 – 7.55 (m, 2H), 7.43 (d, J = 3.9 Hz, 1H), 7.41 – 7.25 (m, 8H), 7.22 (d, J = 3.9 Hz, 1H), 7.10 (t, J = 6.0 Hz, 1H), 4.24 (s, 1H), 3.79 – 3.71 (m, 4H), 3.44 (q, J = 6.3 Hz, 2H), 2.82 (t, J = 6.1 Hz, 2H), 2.71 (t, J = 6.6 Hz, 2H), 1.91 (quintet, J = 6.5 Hz, 2H), 1.80 (quintet, 2H). 13C NMR (101 MHz, CDCl3) δ 162.68, 149.13, 137.49, 135.78,133.66, 129.74, 129.29, 129.18, 128.95, 128.62, 128.29, 126.18, 123.72,63.18, 58.69, 53.57, 50.97, 37.63, 27.34, 25.88. TLC-MS (ESI+): C 24 H 28 The theoretical m / z value for N₂O₂S is 408.19. The measured value is 431.3 [M+Na]. + HPLC t R = 5.528 min.

[0175] N-(3-(benzyl(phenethyl)amino)propyl)-5-phenylthiophene-2-carboxamide (29). Prepared according to general procedure 1 from 5-phenylthiophene-2-carboxylic acid (0.45 mmol) and N 1 -Benzyl-N 1 1,3-Phenethylpropane-1,3-diamine dihydrochloride (29a, 0.45 mmol, 1 eq.). Purified by rapid chromatography using PE / EtOAc (0–60%). Yield: 24 mg (12%), white solid. 1 ¹H NMR (400 MHz, CDCl₃) δ 7.63 – 7.50 (m, 2H), 7.38 – 7.31 (m, 2H), 7.30 – 7.16 (m, 9H), 7.16 – 7.04 (m, 5H), 3.63 (s, 2H), 3.39 (q, J = 5.7 Hz, 2H), 2.86 – 2.70 (m, 4H), 2.61 (t, J = 5.9 Hz, 2H), 1.71 (quintet, J = 6.0 Hz, 2H). 13 C NMR (101 MHz, CDCl3) δ 161.86, 148.42, 140.30, 138.92, 138.20, 133.81,129.44, 129.13, 128.82, 128.79, 128.56, 128.47, 127.42, 126.23, 126.17,123.35, 59.01, 55.52, 52.59, 39.80, 33.04, 25.87. TLC-MS (ESI+): C 29 H 30The theoretical m / z value for N2OS is 454.21. The measured value is 477.1 [M+Na] + HPLC t R = 7.138 min.

[0176] N-(3-(benzyl(3-phenylpropyl)amino)propyl)-5-phenylthiophene-2-carboxamide (30). Prepared according to general procedure 1 from 5-phenylthiophene-2-carboxylic acid (0.42 mmol) and N 1 -Benzyl-N 1 1,3-(3-phenylpropyl)propane-1,3-diamine dihydrochloride (30a, 0.42 mmol, 1 eq.). Purified by rapid chromatography using DCM / MeOH (0–6%). Yield: 19 mg (10%), colorless oil. 1 ¹H NMR (400 MHz, CDCl₃) δ 7.62 (d, 2H), 7.45 – 7.13 (m, 15H), 3.64 (s, 2H), 3.58 – 3.40 (m, 3H), 2.79 – 2.48 (m, 6H), 1.91 (quintet, J = 9.9, 7.5 Hz, 2H), 1.79 (quintet, 2H). 13 C NMR (101 MHz, CDCl3) δ 162.02,148.49, 141.92, 138.58, 138.01, 133.77, 129.51, 129.14, 129.00, 128.59,128.50, 128.41, 127.46, 126.17, 125.98, 123.41, 59.00, 53.30, 52.80, 39.84,33.78, 28.33, 25.70. TLC-MS (ESI+): C 30 H 32 The theoretical m / z value for N2OS is 468.22. The measured value is 491.2 [M+Na]. + HPLC t R = 7.457 min.

[0177] N-(3-(benzyl(ethyl)amino)propyl)-5-(3-fluorophenyl)thiophene-2-carboxamide (31). Prepared from N-(3-(benzyl(ethyl)amino)propyl)-5-bromothiophene-2-carboxamide (31a, 0.29 mmol) and (3-fluorophenyl)boronic acid (0.32 mmol, 1.1 eq.) according to general procedure 2. Purified by rapid chromatography using DCM / MeOH (1–4%). Yield: 29 mg (25%), brown semi-solid. 1 ¹H NMR (400 MHz, CDCl₃) δ 7.98 (s, 1H), 7.39 – 7.25 (m, 8H), 7.24 – 7.21 (m, 1H), 7.18 (d, J = 3.9 Hz, 1H), 7.06 – 6.98 (m, 1H), 3.60 (s, 2H), 3.51 (q, J = 11.2, 4.1 Hz, 2H), 2.69 – 2.57 (m, 4H), 1.78 (quintet, J = 11.6, 5.8 Hz, 2H), 1.11 (t, J = 7.1 Hz, 3H). 13 C NMR (101 MHz, CDCl3) δ164.45, 162.00, 161.65, 146.73, 146.70, 139.15, 135.97, 135.89, 130.75,130.67, 129.62, 128.72, 128.59, 127.48, 124.05, 121.90, 121.87, 115.32,115.11, 113.11, 112.89, 58.59, 52.72, 47.12, 40.58, 25.20, 11.15. TLC-MS (ESI+): C 23 H 25 The theoretical m / z value for FN2OS is 396.17. The measured value is 397.2 [M+H]. + HPLC t R = 6.288 min.

[0178] N-(3-(ethyl(4-fluorobenzyl)amino)propyl)-5-(3-fluorophenyl)thiophene-2-carboxamide (32a) was prepared from 5-bromo-N-(3-(ethyl(4-fluorobenzyl)amino)propyl)thiophene-2-carboxamide (32a, 0.33 mmol) and (3-fluorophenyl)boronic acid (0.36 mmol, 1.1 eq.) according to general procedure 2. Purification was performed by rapid chromatography using DCM / MeOH (1–4%). Yield: 53 mg (40%), brown semi-solid. 1 ¹H NMR (400 MHz, CDCl₃) δ 7.83 (s, 1H), 7.40–7.33 (m, 2H), 7.30–7.23 (m, 4H), 7.20 (d, J = 3.9 Hz, 1H), 7.06–6.99 (m, J = 8.1, 5.7, 3.5 Hz, 1H), 6.99–6.92 (m, 2H), 3.56 (s, 2H), 3.51 (q, J = 11.5, 5.4 Hz, 2H), 2.67–2.50 (m, 4H), 1.78 (quintet, J = 11.7, 5.9 Hz, 2H), 1.10 (t, J = 7.1 Hz, 3H). 13 C NMR (101 MHz, CDCl3) δ 164.45, 163.42, 162.00,161.63, 160.98, 146.82, 146.79, 138.96, 135.88, 135.80, 131.11, 131.03,130.79, 130.71, 129.87, 128.75, 124.07, 121.91, 121.89, 115.50, 115.40,115.29, 115.19, 113.11, 112.89, 57.74, 52.46, 47.02, 40.44, 25.29, 11.16. TLC-MS (ESI+): C 23 H 24 The theoretical m / z value for F2N2OS is 414.16. The measured value is 415.2 [M+H]. + HPLC t R = 6.443min.

[0179] (3-(5-phenylthiophene-2-carbamate)propyl)tert-butyl carbamate (33). Prepared from 5-phenylthiophene-2-carboxylic acid (2.01 mmol) and (3-aminopropyl)carbamate (2.01 mmol, 1 eq.) according to general procedure 1. Purified by filtration and washed with water. Yield: 637 mg (88%), brown solid. 1 H NMR (400 MHz, DMSO) δ8.49 (t, 1H), 7.76 – 7.65 (m, J = 7.9, 5.9 Hz, 3H), 7.53 (d, J = 3.9 Hz, 1H), 7.44 (t, J = 7.6 Hz, 2H), 7.39 – 7.33 (m, J = 7.3 Hz, 1H), 6.80 (t, J = 5.0Hz, 1H), 3.24 (q, J = 13.0, 6.7 Hz, 2H), 2.98 (q, J = 12.9, 6.6 Hz, 2H), 1.63 (quint, J = 6.9 Hz, 2H), 1.38 (s, 9H). 13 C NMR (101 MHz, DMSO) δ 160.87,155.58, 147.15, 139.02, 133.13, 129.20, 128.84, 128.41, 125.59, 124.19,77.48, 39.52, 37.72, 36.88, 29.64, 28.23. TLC-MS (ESI+): C 19 H 24 The theoretical m / z value for N₂O₃S is 360.15. The measured value is 382.8 [M+Na]. + HPLC t R = 8.697 min.

[0180] N-(3-(benzylamino)propyl)-5-phenylthiophene-2-carboxamide (34) was prepared from benzaldehyde (0.51 mmol), N-(3-aminopropyl)-5-phenylthiophene-2-carboxamide hydrochloride (34a, 0.51 mmol, 1 eq.), and triethylamine according to general procedure 3. Purification was performed by rapid chromatography using DCM / MeOH (1–10%). Yield: 53 mg (30%), pale yellow solid. 1¹H NMR (400 MHz, DMSO) δ 8.78 (t, J = 5.8 Hz, 1H), 8.21 (s, 1H), 7.79 (d, J = 3.9 Hz, 1H), 7.75 – 7.65 (m, 2H), 7.56 – 7.30 (m, 8H), 4.02 (s, 2H), 3.32 (q, J = 6.5 Hz, 2H), 2.85 (t, J = 7.4 Hz, 2H), 1.87 (quintet, J = 6.8 Hz, 2H). 13 C NMR (101 MHz, DMSO) δ 161.08, 147.28, 138.82, 134.55, 133.10, 129.41,129.22, 129.15, 128.47, 128.44, 128.17, 125.61, 124.23, 50.80, 44.99, 36.61, 26.83. TLC-MS (ESI+): C 21 H 22 The theoretical m / z value for N2OS is 350.15. The measured value is 372.9 [M+Na] + HPLC t R = 5.935 min.

[0181] N-(3-(3,4-dihydroisoquinoline-2(1H)-yl)propyl)-5-phenylthiophene-2-carboxamide (35a). Prepared from 5-phenylthiophene-2-carboxylic acid (0.53 mmol) and 3-(3,4-dihydroisoquinoline-2(1H)-yl)propyl-1-amine dihydrochloride (35a, 0.53 mmol, 1 eq.) according to general procedure 1. Purified by rapid chromatography using DCM / MeOH / 1% NH3 (1.5–2%). Yield: 17 mg (9%), yellow semi-solid. 1¹H NMR (400 MHz, CDCl₃) δ 8.74 (s, 1H), 7.40 –7.27 (m, 5H), 7.25 – 7.12 (m, 3H), 7.09 – 7.04 (m, J = 7.3 Hz, 1H), 7.02 (d, J = 3.9 Hz, 1H), 6.91 (d, J = 3.9 Hz, 1H), 3.75 (s, 2H), 3.62 (dd, J = 11.0, 5.1 Hz, 2H), 3.00 (t, J = 5.9 Hz, 2H), 2.85 (t, J = 6.0 Hz, 2H), 2.79 (t, 2H), 1.89 (quintet, 2H). 13 C NMR (101 MHz, CDCl3) δ 161.83, 147.98, 138.07,134.31, 134.18, 133.70, 129.15, 128.98, 128.82, 128.27, 126.92, 126.69,126.11, 126.07, 123.25, 58.85, 56.23, 51.52, 41.42, 29.38, 24.24. TLC-MS (ESI+): C 23 H 24 The theoretical m / z value for N2OS is 376.16. The measured value is 399.3 [M+Na] + HPLC t R = 5.191 min.

[0182] N-(3-((3-(4-fluorobenzamido)benzyl)(methyl)amino)propyl)-5-phenylthiophene-2-carboxamide (36a) was prepared from 5-phenylthiophene-2-carboxylic acid (0.34 mmol) and N-(3-(((3-aminopropyl)(methyl)amino)methyl)phenyl)-4-fluorobenzamide (36a, 0.34 mmol, 1 eq.) according to general procedure 1. Purification was performed by rapid chromatography using DCM / MeOH (5–8%). Yield: 96 mg (56%), pale yellow solid. 1H NMR (400 MHz, DMSO) δ 10.35(s, 1H), 8.63 (s, 1H), 8.09 – 8.00 (m, 2H), 7.92 (s, 1H), 7.75 – 7.62 (m,4H), 7.52 (d, J = 3.9 Hz, 1H), 7.44 (dd, J = 10.3, 4.7 Hz, 2H), 7.37 (dd, J =12.3, 5.2 Hz, 4H), 7.18 (s, 1H), 4.02 (s, 1H), 3.55 – 3.22 (m, 3H), 2.33 (s,2H), 1.98 – 1.72 (m, 2H), 1.29 – 0.63 (m, 3H). 13 C NMR (101 MHz, DMSO) δ165.35, 164.48, 162.87, 161.10, 147.30, 139.26, 138.83, 133.11, 131.24,130.45, 130.36, 129.24, 129.06, 128.80, 128.49, 125.62, 124.22, 115.46,115.24, 54.91, 45.66, 36.96, 21.06, 8.60. TLC-MS (ESI+): C 29 H 28 The theoretical m / z value for FN3O2S is 501.19. The measured value is 502.2 [M+H]. + HPLC t R = 6.709 min.

[0183] N-(tert-butyl)-1-(3-(5-phenylthiophene-2-carboxamido)propyl)piperidine-4-carboxamide (37a) was prepared from 5-phenylthiophene-2-carboxylic acid (0.48 mmol) and 1-(3-aminopropyl)-N-(tert-butyl)piperidine-4-carboxamide dihydrochloride (37a, 0.48 mmol, 1 eq.) according to general procedure 1. Purification was performed by rapid chromatography using DCM / MeOH (10–20%). Yield: 940 mg (46%), white solid. 1H NMR (400 MHz, CDCl3) δ 8.14 (t, 1H), 7.70 –7.66 (m, 2H), 7.56 (d, J = 3.9 Hz, 1H), 7.43 – 7.37 (m, 2H), 7.33 – 7.30 (m,J = 3.3, 1.2 Hz, 1H), 7.30 (d, J = 3.9 Hz, 1H), 5.25 (s, 1H), 3.55 (q, J =11.2, 5.4 Hz, 2H), 3.08 (d, J = 11.1 Hz, 2H), 2.53 (t, 2H), 2.04 – 1.82 (m,7H), 1.77 (dt, J = 11.5, 5.9 Hz (2H), 1.33 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ174.1, 162.0, 148.4, 137.9, 133.8, 129.5, 129.2, 128.4, 126.3, 123.6, 58.9,53.7, 51.1, 41.0, 29.2, 29.0, 24.5. TLC-MS (ESI+): C 24 H 33 The theoretical m / z value for N3O2S is 427.23. The measured value is 428.4 [M+H]. + HPLC t R = 5.982 min.

[0184] 3-((5-phenylthiophene-2-carbamate)methyl)piperidin-1-carboxylic acid tert-butyl ester (38). Prepared from 5-phenylthiophene-2-carboxylic acid (1.47 mmol) and 3-(aminomethyl)piperidin-1-carboxylic acid tert-butyl ester (1.47 mmol, 1 eq.) according to general procedure 1. Purified by rapid chromatography using DCM / MeOH (0–1%). Yield: 588 mg (quant.), yellow oil. 1H NMR (400 MHz, CDCl3) δ 7.63 – 7.55 (m, 2H), 7.52 (d, J = 3.9 Hz, 1H), 7.41 – 7.33 (m, J = 7.4 Hz, 2H), 7.33 – 7.27 (m, J = 7.3 Hz, 1H), 7.22 (d, J= 3.9 Hz, 1H), 3.93 – 3.40 (m, J = 46.9 Hz, 3H), 3.35 – 2.98 (m, J = 39.8,28.9 Hz, 2H), 2.97 – 2.74 (m, J = 47.8, 19.7 Hz, 2H), 1.95 – 1.76 (m, 2H),1.70 – 1.56 (m, 1H), 1.44 (s, 9H), 1.41 – 1.18 (m, J = 21.7, 16.4, 7.7 Hz, 2H). 13 C NMR (101 MHz, CDCl3) δ 162.15, 155.28, 148.79, 137.94, 133.67, 129.07,129.07, 128.87, 128.45, 128.45, 126.12, 123.51, 79.71, 47.06, 45.14, 41.58,35.62, 28.51, 28.51, 28.51, 28.19, 23.66. TLC-MS (ESI+): C 22 H 28 The theoretical m / z value for N₂O₃S is 400.18. The measured value is 423.2 [M+Na]. + HPLC t R = 9.442 min.

[0185] N-((1-benzylpiperidin-3-yl)methyl)-5-phenylthiophene-2-carboxamide (39). Prepared from 5-phenylthiophene-2-carboxylic acid (0.5 mmol) and (1-benzylpiperidin-3-yl)methylamine hydrochloride (39a, 0.5 mmol, 1 eq.) according to general procedure 1. Purified by rapid chromatography using DCM / MeOH (1–6%). Yield: 26 mg (14%), yellow oil. 1HNMR (400 MHz, CDCl3) δ 7.60 – 7.48 (m, 2H), 7.43 – 6.97 (m, 10H), 6.63 (s,1H), 3.46 (dd, 2H), 3.37 – 3.19 (m, 2H), 2.82 – 2.42 (m, 2H), 2.27 – 2.00 (m,2H), 1.96 – 1.80 (m, 1H), 1.73 – 1.59 (m, 2H), 1.58 – 1.44 (m, 1H), 1.16 –1.03 (m, 1H). 13 C NMR (101 MHz, CDCl3) δ 162.09, 148.72, 137.94, 137.39,133.71, 129.54, 129.15, 128.97, 128.54, 128.43, 127.44, 126.18, 123.49,63.35, 57.43, 53.90, 43.81, 35.60, 28.26, 24.11. TLC-MS (ESI+): C 24 H 26 The theoretical m / z value for N2OS is 390.18. The measured value is 390.9 [M+H]. + HPLC t R = 5.838 min.

[0186] (4-Benzyl-1,4-diazacycloheptane-1-yl)(5-phenylthiophene-2-yl)methyl ketone (40). Prepared from 2-phenylthiazol-4-carboxylic acid (0.5 mmol) and 1-benzyl-1,4-diazacycloheptane (0.5 mmol, 1 eq.) according to general procedure 1. Purified by rapid chromatography using DCM / MeOH (1–3%). Yield: 152 mg (83%), white solid. 1 H NMR(400 MHz, DMSO) δ 7.70 (d, J = 7.5 Hz, 2H), 7.55 – 7.17 (m, 10H), 3.90 – 3.53(m, J = 65.5 Hz, 6H), 2.79 – 2.57 (m, 4H), 1.88 (s, 2H). 13C NMR (101 MHz, DMSO) δ 162.66, 146.27, 138.99, 137.51, 132.94, 130.27, 129.25, 128.55,128.45, 128.21, 126.91, 125.68, 123.66, 61.08, 54.42, 48.38, 45.81, 38.25, 28.27. TLC-MS (ESI+): C 23 H 24 The theoretical m / z value for N2OS is 376.16. The measured value is 377.2 [M+H]. + HPLC t R = 5.182 min.

[0187] 4-Fluoro-N-(3-((4-(5-phenylthiophene-2-carbonyl)-1,4-diazacycloheptane-1-yl)methyl)phenyl)benzamide (41). Prepared from 5-phenylthiophene-2-carboxylic acid (0.27 mmol) and N-(3-((1,4-diazacycloheptane-1-yl)methyl)phenyl)-4-fluorobenzamide hydrochloride (41a, 0.27 mmol, 1 eq.) according to general procedure 1. Purified by rapid chromatography using DCM / MeOH (1–3%). Yield: 80 mg (57%), pale yellow solid. 1 ¹H NMR (400 MHz, CDCl₃) δ 8.07 (s, 1H), 7.85 (s, 2H), 7.70 – 7.53 (m, J = 6.9 Hz, 4H), 7.42 – 7.36 (m, J = 7.4 Hz, 2H), 7.35 – 7.26 (m, 3H), 7.22 (s, 1H), 7.15 – 7.05 (m, 3H), 3.79 (s, 4H), 3.65 (s, 2H), 2.84 – 2.74 (m, J = 13.7, 8.7 Hz, 2H), 2.70 (s, 2H), 1.96 (quintet, 2H). 13C NMR (101 MHz, CDCl3) δ 166.24, 164.94, 164.36,163.73, 138.25, 137.07, 133.59, 131.23, 131.20, 130.19, 129.74, 129.66,129.18, 129.15, 128.48, 126.19, 125.11, 122.80, 120.77, 119.46, 115.97,115.75, 62.27, 54.93, 49.71, 49.05, 46.55, 29.82. TLC-MS (ESI+): C 30 H 28 The theoretical m / z value for FN3O2S is 513.19. The measured value is 536.2 [M+Na]. + HPLC t R = 6.447 min.

[0188] N-(3-nitrobenzyl)-5-phenylthiophene-2-carboxamide (42). Prepared from 5-phenylthiophene-2-carboxylic acid (0.98 mmol) and (3-nitrophenyl)methylamine hydrochloride (42a, 0.98 mmol, 1 eq.) according to general procedure 1. Purified by rapid chromatography using DCM / MeOH (0–1%). Yield: 230 mg (69%), white solid. 1 H NMR (400 MHz, CDCl3) δ 8.19 (s, 1H), 8.11 (d, J = 8.2, 1.3 Hz, 1H), 7.72 (d, J = 7.8 Hz,1H), 7.64 – 7.57 (m, 2H), 7.54 (d, J = 3.9 Hz, 1H), 7.50 (t, J = 7.9 Hz, 1H), 7.44 – 7.30 (m, 3H), 7.26 (d, J = 3.8 Hz, 1H), 6.73 (t, J = 5.6 Hz, 1H), 4.71(d, J = 6.1 Hz, 2H). 13 C NMR (101 MHz, CDCl3) δ 162.24, 149.74, 148.58, 140.63,136.67, 134.09, 133.45, 129.85, 129.70, 129.24, 128.85, 126.28, 123.69,122.71, 122.60, 43.30. TLC-MS (ESI+): C 18H 14 The theoretical m / z value for N₂O₃S is 338.07. The measured value is 361.1 [M+Na]. + HPLC t R = 8.347 min.

[0189] N-(3-(4-fluorobenzamido)benzyl)-5-phenylthiophene-2-carboxamide (43a) was prepared from 4-fluorobenzoic acid (0.5 mmol) and N-(3-aminobenzyl)-5-phenylthiophene-2-carboxamide (43a, 0.5 mmol, 1 eq.) according to general procedure 1. The mixture was purified by filtration and washed with water. Yield: 187 mg (87%), white solid. 1 H NMR (400 MHz, DMSO) δ 10.29 (s, 1H), 9.12 (s, 1H), 8.36 – 6.84 (m, J = 183.8, 132.7, 90.8Hz, 15H), 4.38 (d, J = 90.2 Hz, 2H). 13 C NMR (101 MHz, DMSO) δ 165.28, 164.43,162.80, 160.96, 147.51, 139.97, 139.19, 138.83, 133.12, 131.34, 130.43,130.34, 129.23, 128.56, 128.49, 125.66, 124.32, 122.73, 119.16, 119.01,115.37, 115.16, 42.55. TLC-MS (ESI+): C 25 H 19 The theoretical m / z value for FN₂O₂S is 430.12. The measured value is 453.2 [M+Na]. + HPLC t R = 8.786 min.

[0190] N-(2-(1-methylpyrrolidone-2-yl)ethyl)-2-phenylthiazol-4-carboxamide (44). Prepared from 2-phenylthiazol-4-carboxylic acid (0.39 mmol) and 2-(1-methylpyrrolidone-2-yl)ethyl-1-amine (0.39 mmol, 1 eq.) according to general procedure 1. Purified by rapid chromatography using DCM / MeOH (10%). Yield: 48 mg (39%), light brown semi-solid. 1H NMR (400 MHz, CDCl3) δ 8.04 (s, 1H), 7.98 – 7.87 (m, 3H), 7.46 – 7.40 (m,3H), 3.73 – 3.39 (m, 3H), 3.08 – 2.93 (m, 1H), 2.80 – 2.71 (m, J = 10.3 Hz,1H), 2.69 (s, 3H), 2.39 – 2.02 (m, 4H), 1.99 – 1.84 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 168.42, 161.65, 150.51, 132.79, 130.78, 129.15, 126.73, 123.04, 66.48, 56.16, 39.35, 36.57, 31.07, 29.79, 21.73. TLC-MS (ESI+): C 17 H 21 The theoretical m / z value for N3OS is 315.14. The measured value is 316.2 [M+H]. + HPLC t R = 3.921 min.

[0191] 4-(2-phenylthiazol-4-carbonyl)-1,4-diazacycloheptane-1-carboxylic acid tert-butyl ester (45). Prepared from 2-phenylthiazol-4-carboxylic acid (2.44 mmol) and 1,4-diazacycloheptane-1-carboxylic acid tert-butyl ester (2.44 mmol, 1 eq.) according to general procedure 1. Purified by rapid chromatography using PE / EtOAc (10–30%). Yield: 784 mg (83%), yellow oil. 1 H NMR (400 MHz, DMSO) δ 8.27 – 8.09 (m, J = 26.9, 12.4 Hz, 1H), 8.04 – 7.91 (m, J = 3.4 Hz, 2H), 7.66 – 7.45 (m, J = 4.1 Hz, 3H), 3.95 – 3.36(m, 8H), 1.99 – 1.72 (m, J = 32.8, 15.9 Hz, 2H), 1.47 – 1.20 (m, J = 35.3 Hz, 9H). 13C NMR (101 MHz, DMSO) δ 166.4, 163.1, 154.4, 151.0, 132.6, 130.7, 129.3,126.3, 126.2, 78.7, 45.3, 34.9, 28.3, 28.0, 27.8, 26.0. TLC-MS (ESI+): C 20 H 25 The theoretical m / z value for N3O3S is 387.16. The measured value is 410.2 [M+Na]. + HPLC t R = 8.982 min.

[0192] 4-(3-(2-phenylthiazol-4-carboxamido)propyl)piperazine-1-carboxylic acid tert-butyl ester (46a). Prepared according to general procedure 1 from 2-phenylthiazol-4-carboxylic acid (0.43 mmol) and 4-(3-aminopropyl)piperazine-1-carboxylic acid tert-butyl ester (46a, 0.43 mmol, 1 eq.). Purified by filtration and washed with water. Yield: 128 mg (69%), white solid. 1 ¹H NMR (400MHz, DMSO) δ 8.65 (t, 1H), 8.28 (s, 1H), 8.02 (s, 2H), 7.52 (s, 3H), 3.43–3.32 (m, J = 16.8 Hz, 6H), 2.47–2.20 (m, 6H), 1.71 (quintet, 2H), 1.39 (d, J = 17.7 Hz, 9H). 13 C NMR (101 MHz, DMSO) δ 167.2, 160.2, 153.8, 150.9, 132.5,130.7, 129.2, 126.4, 123.9, 78.7, 56.2, 52.6, 37.9, 28.0, 25.9. TLC-MS (ESI+): C 22 H 30 The theoretical m / z value for N4O3S is 430.20. The measured value is 431.2 [M+H]. + HPLC t R = 5.651 min.

[0193] 4-(3-(((2-phenylthiazol-4-yl)methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester (47). Prepared from 2-phenylthiazol-4-carboxaldehyde (0.47 mmol, 1.1 eq.) and 4-(3-aminopropyl)piperazine-1-carboxylic acid tert-butyl ester (46a, 0.43 mmol) according to general procedure 3. Purified by rapid chromatography using DCM / MeOH (5–8%). Yield: 13 mg (7%), yellow oil. 1 ¹H NMR (400 MHz, CDCl₃) δ 7.94 – 7.81 (m, J = 6.3, 2.7 Hz, 2H), 7.54 (s, 1H), 7.48 – 7.40 (m, J = 6.7, 3.5 Hz, 3H), 4.26 (s, 2H), 3.41 (s, 4H), 3.20 (s, 2H), 2.49 (d, J = 33.3 Hz, 6H), 2.00 (quintet, 2H), 1.42 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 169.9, 154.6, 133.0, 130.8, 129.2, 126.7,119.7, 80.0, 58.1, 48.8, 46.8, 29.8, 28.5, 22.1, 14.2. TLC-MS (ESI+): C 22 H 32 The theoretical m / z value for N4O2S is 416.22. The measured value is 417.2 [M+Na]. + HPLC t R = 3.448 min.

[0194] N-Benzyl-1-((2-phenylthiazol-4-yl)methyl)piperidine-4-carboxamide (48). Prepared from 2-phenylthiazol-4-carboxaldehyde (0.39 mmol, 1 eq.) and N-benzylpiperidine-4-carboxamide hydrochloride (48a, 0.48 mmol) according to general procedure 3. Purified by rapid chromatography using DCM / MeOH (2–7%). Yield: 99 mg (71%), white solid. 1H NMR(400 MHz, CDCl3) δ 8.00 – 7.89 (m, 2H), 7.45 – 7.37 (m, 3H), 7.35 – 7.22 (m,5H), 7.13 (s, 1H), 5.76 (t, 1H), 4.44 (d, J = 5.6 Hz, 2H), 3.73 (d, J = 0.6Hz, 2H), 3.07 (dt, J = 8.9, 3.0 Hz, 2H), 2.21 – 2.04 (m, 3H), 1.92 – 1.75 (m,4H). 13 C NMR (101 MHz, CDCl3) δ 174.9, 168.0, 155.2, 138.5, 133.9, 130.0,129.0, 128.9, 127.9, 127.7, 126.7, 115.9, 58.7, 53.4, 43.6, 43.5, 29.2. TLC-MS(ESI+):C 23 H 25 The theoretical m / z value for N3OS is 391.17. The measured value is 392.2 [M+H]. + HPLC t R = 5.446 min.

[0195] 4-(2-(benzo[b]thiophene-2-carboxamido)ethyl)piperazine-1-carboxylic acid tert-butyl ester (49). Prepared from benzo[b]thiophene-2-carboxylic acid (0.39 mmol) and 4-(2-aminoethyl)piperazine-1-carboxylic acid tert-butyl ester (0.39 mmol, 1 eq.) according to general procedure 1. Purified by rapid chromatography using DCM / MeOH (0–3%). Yield: 7 mg (46%), yellow oil. 1 H NMR (400 MHz, CDCl3) δ 7.88 – 7.80 (m, J = 10.3, 4.2 Hz, 2H), 7.77 (s,1H), 7.49 – 7.33 (m, 2H), 6.95 (t, 1H), 3.57 (q, J = 11.3, 5.6 Hz, 2H), 3.52– 3.41 (m, 4H), 2.63 (t, J = 6.0 Hz, 2H), 2.55 – 2.39 (m, 4H), 1.46 (s, 9H). 13C NMR (101 MHz, CDCl3) δ 162.4, 154.8, 140.9, 139.2, 138.6, 126.4, 125.3, 125.1, 125.0, 122.8, 80.0, 56.5, 52.7, 36.4, 28.5. TLC-MS (ESI+): C 20 H 27 The theoretical m / z value for N3O3S is 389.18. The measured value is 412.2 [M+Na]. + HPLC t R = 5.225 min.

[0196] Measurement

[0197] ACKR3 β-inhibitor protein recruitment assay. In HEK293T cells, the potency and efficacy of compounds 1 to 49 were evaluated by ligand-induced recruitment of β-repressor protein to ACKR3 using the NanoLuc complementation assay (NanoBiT, Promega Corporation, Madison, WI, USA). (Szpakowska, M.; Nevins, AM; Meyrath, M.; Rhainds, D.; D'huys, T.; Guité-Vinet, F.; Dupuis, N.; Gauthier, P.-A.; Counson, M.; Kleist, A.; St-Onge, G.; Hanson, J.; Schols, D.; Volkman, BF; Heveker, N.; Chevigné, A.) Different contributions of chemokine n-terminal features attest to a different ligand binding mode and a bias towards activation of ACKR3 / CXCR7 compared with CXCR4 and CXCR3. (Different contributions from the n-terminal features of chemokines confirm different ligand binding patterns compared to CXCR4 and cxcr3, with a bias towards ACKR3 / CXCR7 activation). Br. J. Pharmacol. 2018, 175, 1419-1438; Dixon, AS; Schwinn, MK; Hall, MP; Zimmerman, K.; Otto, P.; Lubben, TH; Butler, BL; Binkowski, BF; Machleidt, T.; Kirkland, TA; Wood, MG; Eggers, CT; Encell, LP; Wood, KV. Nanoluc complementation reporter optimized for accurate measurement of protein interactions in cells. ACS Chem. Biol. 2016, 11, 400-408; Meyrath, M.; Palmer, CB.Reynders, N.; Vanderplasschen, A.; Ollert, M.; Bouvier, M.; Szpakowska, M.; Chevigné, A. Proadrenomedullin n-terminal 20 peptides (pamps) are agonists of the chemokine scavenger receptor ACKR3 / CXCR7. ACS Pharmacology & Translational Science 2021, 4, 813-823; and Szpakowska, M.; Meyrath, M.; Reynders, N.; Counson, M.; Hanson, J.; Steyaert, J.; Chevigné, A. Mutational analysis of the extracellular disulphide bridges of the atypical chemokine receptor. ACKR3 / CXCR7 uncovers multiple binding and activation modes for its chemokine and endogenous non-chemokine agonists. (Biochem. Pharmacol. 2018, 153, 299-309). In summary: 5 × 10⁶ HEK293T cells were seeded in 10⁻⁶ cm culture dishes and co-transfected with the pNBe vector (C-terminus fused to SmBiT) 24 h later. The pNBe vector encodes either the human ACKR3 or CXCR4 receptor (C-terminus fused to SmBiT) or β-repressor-1 (N-terminus fused to LgBiT). After 24 h, transfected cells were harvested, incubated at 37 °C for 15 min with Nano-Glo live cell substrate diluted 200-fold, and aliquoted into white 96-well plates (5 × 10⁴ cells / well). The prepared cells were then used at a concentration of 0.Compounds ranging from 3 nM to 100 µM were used for treatment. Induction of recruitment to the receptor's β-repressor protein was evaluated by measuring bioluminescence using a Mithras LB940 luminometer (Berthold Technologies). To determine the efficacy of the compounds and to generate concentration-response curves, the recorded signals were compared with those of CXCL12 (E), considered a complete agonist reference ligand, at 300 nM. max Compare the values ​​of (= 100%).

[0198] All curves were fitted to data points derived from the average of at least three independent experiments. Results with a p-value < 0.05 were considered statistically significant.

[0199] Flow cytometry assay. Inhibition of platelet degranulation was determined by flow cytometry. Platelets were treated with collagen-related peptide (CRP-XL, CambCol Laboratories, Cambridge, UK), and platelet activation status was measured (Geue, S.; Aurbach, K.; Manke, M.-C.; Manukjan, G.; Münzer, P.; Stegner, D.; Brähler, C.; Walker-Allgaier, B.; Märklin, M.; Borst, CE; Quintanilla-Fend, L.; Rath, D.; Geisler, T.; Salih, h. R.; Seizer, P.; Lang, F.; Nieswandt, B.; Gawaz, M.; Schulze, H.; Pleines, I.; Borst, O.). Pivotal role of pdk1 in megakaryocyte cytoskeletal dynamics and polarization during platelet biogenesis. (The crucial role of pdk1 in megakaryocyte cytoskeleton dynamics and polarization during platelet biogenesis). Blood 2019, 134, 1847-1858; and Malehmir, M.; Pfister, D.; Gallage, S.; Szydlowska, M.; Inverso, D.; Kotsiliti, E.; Leone, V.; Peiseler, M.; Surewaard, BGJ; Rath, D.; Ali, A.; Wolf, MJ; Drescher, H.; Healy, ME; Dauch, D.; Kroy, D.; Krenkel, O.; Kohlhepp, M.; Engleitner, T.; Olkus, A.; Sijmonsma, T.; Volz, J.; Deppermann, C.; Stegner, D.; Helbling, P.; Nombela-Arrieta, C.; Rafiei, A.; Hinterleitner, M.; Rall, M.; Baku, F.; Borst, O.; Wilson, CL; Leslie, J.; O'Connor, T.; Weston, CJ; Chauhan, A.; Adams, DH; Sheriff, L.Teijeiro, A.; Prinz, M.; Bogeska, R.; Anstee, N.; Bongers, MN; Notohamiprodjo, M.; Geisler, T.; Withers, DJ; Ware, J.; Mann, DA; Augustin, h. G.; Vegiopoulos, A.; Milsom, MD; Rose, AJ; Lalor, PF; Llovet, JM; Pinyol, R.; Tacke, F.; Rad, R.; Matter, M.; Djouder, N.; Kubes, P.; Knolle, PA; Unger, K.; Zender, L.; Nieswandt, B.; Gawaz, M.; Weber, A.; Heikenwalder, M. Platelet gpibα is a mediator and potential interventional target for nash and subsequent liver cancer (Platelet gpibα is a mediator and potential intervention target for Nash and secondary hepatocellular carcinoma. Nat. Med. 2019, 25, 641-655). Briefly, PRP was isolated from human peripheral blood collected in adenine-containing citrate-phosphate-glucose solution (CPDA) and diluted with phosphate-buffered saline (PBS) supplemented with CaCl2 and MgCl2 (Sigma Aldrich Co., Ltd., St. Luis, Missouri, USA). PRP (106 platelets / sample) was pre-incubated with the appropriate compound (100 μM) for 15 min. The samples were then treated with 1 μg / ml CRP-XL (CambCol Laboratories, Cambridge, UK) for 30 min at room temperature in the presence of a fluorescent dye conjugated antibody against P-selectin (CD62P-FITC; Beckman Coulter, Brea, CA, USA). The samples were then fixed with 0.5% paraformaldehyde and analyzed using a FACS-Calibur flow cytometer (FACS-Caliber flow cytometer, Becton-Dickinson, East Rutherford, NJ, USA). Mean fluorescence intensity (MFI) was used as a quantitative measure of platelet surface expression.

[0200] Cell viability assay. RealTime-Glo™ MT cell viability assays were performed using THP1 and HEK293 cells as specified in the manufacturer's manual (Promega Corporation, Madison, WI, USA) to measure the cytotoxicity of the compounds to the cells. Briefly, cells were seeded (5 x 10³ cells / well) in white solid 96-well plates and incubated with the corresponding compound diluted in DMEM (Gibco, Carlsbad, CA, USA). Cell viability reagent was added after seeding. Cells were then incubated in medium containing the compound and viability reagent at 37°C, 5% CO2 for 72 h. The luminescence signal was measured at 1, 24, 48, and 72 hours using a GloMax®-Multi Detection System microplate reader (Promega Corporation, Madison, WI, USA).

[0201] Metabolic stability assays. Confluent liver microsomes from male mice and male humans were purchased from Sekisui XenoTech, LLC, Kansas City, KS, USA. Metabolic stability assays were performed in the presence of an NADPH regeneration system consisting of 5 mM glucose-6-phosphate, 5 U / mL glucose-6-phosphate dehydrogenase, and 1 mM NADP. + Composition. Liver microsomes (20 mg / mL), an NADPH regeneration system, and 4 mM MgCl₂·6H₂O in 0.1 M TRIS-HCl buffer (pH 7.4) were preheated on a shaker at 37°C and 750 rpm for 5 min. The reaction was initiated by adding 10 mM of the preheated compound (resulting in a final concentration of 0.1 mM). At selected time points (0, 10, 20, 30, 60, and 120 min), the reaction was quenched by adding 100 μL of acetonitrile solution of the internal standard (ketoprofen) at concentrations of 600 μM (compound 26), 800 μM (compound 27), and 450 μM (compound 35), respectively. The samples were vortexed for 30 s and centrifuged (relative centrifugal force 21910, 4°C, 20 min). The supernatant was used directly for LC-MS analysis.

[0202] All compounds were incubated in at least triplicate. In addition, negative controls (containing BSA (20 mg / mL) instead of liver microsomes) and positive controls (using verapamil instead of the compounds) were performed. The limit for organic solvents was not exceeded 1% during incubation. Separation and detection of samples were performed on an Alliance 2695 Separations Module HPLC system (Waters Corporation, Milford, MA, USA) equipped with a Phenomenex Kinetex 2.6 μm XB-C18100 Å 50 x 3 mm column (Phenomenex Inc., Torrance, CA, USA), coupled with an Alliance 2996 photodiode array detector and a MICROMASS QUATTRO micro API mass spectrometer (both from Waters Corporation, Milford, MA, USA), using electrospray ionization in positive ion mode. Mobile phase A: 90% water, 10% acetonitrile, and an additional 0.1% formic acid (v / v); Mobile phase B: 100% acetonitrile and an additional 0.1% formic acid (v / v). The gradient was set as follows: 0–2.5 min 0% B, 2.5–10 min 0–40% B, 10–12 min 40% B, 12–12.01 min 40% B to 0% B, 12.01–17 min 0% B, with a flow rate of 0.7 mL / min. The sample was maintained at 10 °C, and the column temperature was set to 20 °C, with an injection volume of 5 μL. The spray voltage, cone voltage, extraction voltage, and RF lens voltage were set to 4 kV, 30 V, 8 V, and 2 V, respectively. The source temperature and desolvation temperature were set to 120 °C and 350 °C, respectively, and the desolvation gas flow rate was set to 750 L / h. Data analysis was performed using MassLynx 4.1 software (Waters Corporation, Milford, MA, USA).

[0203] Solubility determination. A 100 mL flask containing phosphate-buffered saline was prepared using NaCl, KCl, Na₂HPO₄, KH₂PO₄, and distilled water. The buffer was adjusted to pH 7.4 with HCl. A 10 mM stock solution of the compound in DMSO was then prepared. 10 µL of the stock solution was added to 990 µL of PBS in an Eppendorf tube. As a reference, 10 µL of the stock solution was added to 990 µL of HPLC-grade MeOH in an Eppendorf tube. All tubes were centrifuged (16000 relative centrifugation force, 4°C, 20 min). Subsequently, 600 µL of the supernatant was carefully extracted and transferred to an HPLC vial. The area under the curve (AUC) of the compounds was determined by RP-HPLC using an Agilent 1100 Series LC equipped with a Phenomenex Luna C8 analytical column (150 x 4.6 mm, 5 μm), and detected by a UV DAD detector at wavelengths of 254 nm and 230 nm. The solubility of the compounds was calculated by dividing the AUC of the compound in PBS by the AUC of the reference solution and then multiplying by the concentration of the compound in the solution.

[0204] Aggregation assay. For the light transmission aggregation assay, citrate-anticoagulated blood from healthy donors was used. To obtain platelet-rich plasma, whole blood was centrifuged at 210 x for 20 min at room temperature. One x 10^8 platelets were pre-incubated with 100 µM MACKR3 agonist at 37°C for 15 min. Subsequently, the platelets were activated with 2.5 µM ADP, and aggregation was analyzed using an Aggregometer 490-X (Chrono-Log Corp., Havertown, Pennsylvania, United States) at 1,000 rpm and 37°C for 5 min. The maximum platelet aggregation rate and area under the curve were quantified using Aggrolink8 software (Chrono-Log Corp., Havertown, Pennsylvania, United States).

[0205] Synthesis of intermediates

[0206] intermediate N 1 -Benzyl-N 1Synthesis of 1,3-diamine dihydrochloride (1a) of methylpropane. 0.33 g (1.2 mmol) of 1b was dissolved in 6 mL (23.7 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 3 h. The HCl salt of the product formed was filtered off by suction filtration and dried under reduced pressure to give a yellow solid in 72% yield (0.215 g, 0.86 mmol). This product was used without further purification. 1 ¹H NMR (400 MHz, DMSO) δ 11.24 (s, 1H), 8.31 (s, 3H), 7.69 – 7.60 (m, 2H), 7.48 – 7.41 (m, 3H), 4.41 – 4.18 (m, J = 18.8, 13.0, 5.2 Hz, 2H), 3.24 – 3.03 (m, 2H), 2.94 – 2.79 (m, 2H), 2.61 (d, J = 4.9 Hz, 3H), 2.10 (quintet, 2H). 13 C NMR (101 MHz, DMSO) δ131.50, 129.87, 129.46, 128.76, 58.15, 51.69, 38.50, 36.24, 21.61. TLC-MS (ESI+): C 11 H 18 The theoretical m / z value for N2 is 178.15. The measured value is 179.2 [M+H] + .

[0207] N 1 -(4-Methoxybenzyl)-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (18a). 0.315 g (1.02 mmol) of 18b was dissolved in 5.1 mL (20.4 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 3 h. The resulting product was filtered off and dried under reduced pressure to give a white solid in 90% yield (0.258 g, 0.92 mmol). This product was used without further purification. 1¹H NMR (400 MHz, DMSO) δ 11.14 (s, 1H), 8.34 (s, 3H), 7.56 (d, J = 8.7 Hz, 2H), 6.98 (d, J = 8.7 Hz, 2H), 4.34 –4.10 (m, J = 18.2, 13.0, 4.7 Hz, 2H), 3.77 (s, 3H), 3.24 – 2.96 (m, 2H), 2.95– 2.77 (m, J = 12.1, 6.4 Hz, 2H), 2.58 (d, J = 4.4 Hz, 3H), 2.10 (quintet, 2H). 13 C NMR (101 MHz, DMSO) δ 159.99, 133.02, 121.56, 114.06, 57.64, 55.23, 51.32, 38.21, 36.22, 21.61. TLC-MS (ESI+): C 12 H 20 The theoretical m / z value for N₂O is 208.16. The measured value is 209.1 [M+H] + .

[0208] N 1 -(4-Bromobenzyl)-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (19a). 0.354 g (0.99 mmol) of 19b was dissolved in 5 mL (19.8 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 3 h. The resulting product was filtered off and dried under reduced pressure to give a white solid in 98% (0.32 g, 0.97 mmol). This product was used without further purification. 1 ¹H NMR (400 MHz, DMSO) δ 11.38 (s, 1H), 8.30 (s, 3H), 7.71 – 7.58 (m, 4H), 4.42 – 4.18 (m, J = 18.7, 13.1, 5.1 Hz, 2H), 3.27 – 2.98 (m, 2H), 2.94 – 2.77 (m, 2H), 2.61 (d, J = 4.8 Hz, 3H), 2.10 (quintet, 2H). 13C NMR (101 MHz, DMSO) δ 133.73, 131.68, 129.26, 123.10, 57.30, 51.61, 38.47, 36.19, 21.57. TLC-MS (ESI+): C 11 H 17 The theoretical m / z value for BrN2 is 256.06. The measured values ​​are 257.0 / 259.0 [M+H]. + .

[0209] N 1 -(4-Fluorobenzyl)-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (20a). 0.313 g (1.06 mmol) of 20b was dissolved in 5.3 mL (21.1 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 3 h. The HCl salt of the resulting product was filtered off by suction filtration and dried under reduced pressure to give a white solid. This crude product was used without further purification.

[0210] N 1 -(2-Fluorobenzyl)-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (21a). 0.43 g (1.45 mmol) of 21b was dissolved in 7.25 mL (29 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 3 h. The resulting product was filtered off and dried under reduced pressure to give a white semi-solid in 86% yield (0.336 g, 1.25 mmol). This product was used without further purification. 1 H NMR (400 MHz, DMSO) δ 11.32 (s, 1H), 8.34 (s, 3H), 7.83 (td, J = 7.6, 1.4 Hz, 1H), 7.65 – 7.44 (m, J = 7.3, 1.6Hz, 1H), 7.39 – 7.19 (m, 2H), 4.45 – 4.20 (m, J = 18.9, 13.4, 4.6 Hz, 2H), 3.33 – 3.06 (m, 2H), 2.98 – 2.77 (m, 2H), 2.65 (d, J = 4.6 Hz, 3H), 2.23 –2.03 (m, 2H). 13C NMR (101 MHz, DMSO) δ 161.09, 134.10, 132.29, 124.85, 116.85, 115.85, 51.84, 51.22, 38.68, 36.17, 21.75. TLC-MS (ESI+): C 11 H 17 The theoretical m / z value for FN2 is 196.14. The measured value is 197.1 [M+H] + .

[0211] N 1 -(3-Fluorobenzyl)-N 1 1,3-Methylpropane-1,3-diamine dihydrochloride (22a). 0.35 g (1.18 mmol) of 22b was dissolved in 5.9 mL (23.6 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 3 h. The resulting product was filtered off and dried under reduced pressure to give a white solid in 97% yield (0.307 g, 1.14 mmol). This product was used without further purification. 1 ¹H NMR (400 MHz, DMSO) δ 11.40 (s, 1H), 8.29 (s, 3H), 7.61 (d, J = 8.4 Hz, 1H), 7.55 – 7.43 (m, 2H), 7.39 – 7.23 (m, J = 9.0, 5.9, 3.7 Hz, 1H), 4.51 – 4.18 (m, 2H), 3.29 – 2.99 (m, 2H), 2.98 – 2.76 (m, 2H), 2.62 (s, 3H), 2.11 (quintet, 2H). 13 C NMR (101 MHz, DMSO) δ161.90, 132.50, 130.79, 127.68, 118.27, 116.42, 57.44, 51.80, 38.57, 36.21,21.60. TLC-MS (ESI+): C 11 H 17 The theoretical m / z value for FN2 is 196.14. The measured value is 197.1 [M+H] + .

[0212] N 1 -Benzyl-N 11,3-Ethylpropane-1,3-diamine dihydrochloride (23a). 0.28 g (0.96 mmol) of 23b was dissolved in 4.8 mL (19.2 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 3 h. The HCl salt of the resulting product was filtered off by suction filtration and dried under reduced pressure to give a white solid. This crude product was used without further purification.

[0213] N 1 -Benzyl-N 1 -Isopropylpropane-1,3-diamine dihydrochloride (24a). 0.205 g (0.67 mmol) of 24b was dissolved in 3.34 mL (13.4 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 1 h. The resulting product was filtered off by suction filtration and dried under reduced pressure. This product was used without further purification.

[0214] N 1 -Benzyl-N 1 1,3-Cyclopropylpropane-1,3-diamine dihydrochloride (25a). 0.357 g (1.17 mmol) of 25b was dissolved in 5.85 mL (23.45 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 1 h. The resulting product was filtered off by vacuum filtration and dried under reduced pressure. This product was used without further purification. TLC-MS (ESI+): C 13 H 20 The theoretical m / z value for N2 is 204.16. The measured value is 205.2 [M+H] + .

[0215] N 1 -Benzyl-N 1 1,3-Propylpropane-1,3-diamine dihydrochloride (26a). 0.37 g (1.21 mmol) of 26b was dissolved in 6.04 mL (24.15 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 1 h. The resulting product was filtered off by vacuum filtration and dried under reduced pressure to give a colorless oil. This product was used without further purification. TLC-MS (ESI+): C 13 H 22 The theoretical m / z value for N2 is 206.18. The measured value is 207.2 [M+H] + .

[0216] N 1 -Benzyl-N 11,3-Butylpropane-1,3-diamine dihydrochloride (27a). 0.375 g (1.17 mmol) of 27b was dissolved in 5.85 mL (23.4 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 1 h. The resulting product was filtered off by suction filtration and dried under reduced pressure. This product was used without further purification. TLC-MS (ESI+): C 14 H 24 The theoretical m / z value for N2 is 220.19. The measured value is 221.2 [M+H] + .

[0217] 3-((3-aminopropyl)(benzyl)amino)prop-1-ol dihydrochloride (28a). 0.388 g (1.2 mmol) of 28b was dissolved in 6 mL (24 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred overnight at room temperature. The resulting product was dried under reduced pressure to give a yellow oil in 99% (0.353 g, 1.2 mmol). This product was used without further purification. 1 H NMR (400 MHz, DMSO) δ 11.09 (s, 1H), 8.24 (s, 3H), 7.75 – 7.61 (m, 2H), 7.49 – 7.39 (m, 3H), 4.38 – 4.24 (m, 2H), 3.49 – 3.36(m, 2H), 3.22 – 2.79 (m, 7H), 2.20 – 1.97 (m, 2H), 2.00 – 1.81 (m, 2H). 13 C NMR (101 MHz, DMSO) δ 131.39, 129.83, 129.41, 128.75, 57.90, 55.81, 49.25, 48.85, 36.25, 25.76, 21.16. TLC-MS (ESI+): C 13 H 22 The theoretical m / z value for N₂O is 222.17. The measured value is 223.1 [M+H] + .

[0218] N 1 -Benzyl-N 11,3-Phenethylpropane-1,3-diamine dihydrochloride (29a). 0.178 g (0.48 mmol) of 29b was dissolved in 2.4 mL (9.66 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred overnight at room temperature. The resulting product was filtered off by suction filtration and dried under reduced pressure to give a pale brown solid in 99% (0.164 g, 0.48 mmol). This product was used without further purification. 13 C NMR (101 MHz, CDCl3) δ 136.27, 131.57,130.01, 129.39, 129.05, 128.92, 128.84, 127.14, 56.98, 52.71, 51.21, 37.89,29.72, 22.35. TLC-MS (ESI+): C 18 H 24 The theoretical m / z value for N2 is 268.19. The measured value is 269.1 [M+H] + HPLC t R = 1.907 min.

[0219] N 1 -Benzyl-N 1 1,3-(3-phenylpropyl)propane-1,3-diamine dihydrochloride (30a). 0.184 g (0.48 mmol) of 30b was dissolved in 2.4 mL (9.6 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred overnight at room temperature. The resulting product was filtered off by suction filtration and dried under reduced pressure to give a pale brown solid in 99% (0.168 g, 0.47 mmol). This product was used without further purification. 1 H NMR (400 MHz, DMSO) δ 8.29 (s,3H), 7.65 (dd, J = 7.3, 2.2 Hz, 2H), 7.48 – 7.36 (m, 3H), 7.35 – 7.10 (m,6H), 4.39 – 4.22 (m, 2H), 3.24 – 3.07 (m, 2H), 3.03 – 2.90 (m, 2H), 2.90 –2.79 (m, 2H), 2.64 – 2.54 (m, 2H), 2.26 – 1.95 (m, 4H). 13C NMR (101 MHz, DMSO)δ 140.36, 131.38, 129.74, 129.34, 128.70, 128.32, 128.28, 126.06, 55.65,50.66, 48.95, 36.23, 31.98, 24.01, 21.22. TLC-MS (ESI+): C 19 H 26 The theoretical m / z value for N2 is 282.21. The measured value is 283.3 [M+H] + HPLC t R = 2.399 min.

[0220] N-(3-(benzyl(ethyl)amino)propyl)-5-bromothiophene-2-carboxamide (31a). 0.112 g (0.54 mmol) of 5-bromothiophene-2-carboxylic acid was dissolved in 2.2 mL of anhydrous DMF, and 0.227 g (0.6 mmol) of HATU was slowly added. After stirring for 15 min, 0.144 g (0.54 mmol) of 23a and 0.43 mL (2.44 mmol) of DIPEA were added to the mixture. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was diluted with DCM and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na2SO4, filtered, and evaporated to dryness. The mixture was purified by rapid chromatography (SiO2; DCM / MeOH 1 - 4%). The crude product was used without further purification. TLC-MS (ESI+): C 17 H 21 The theoretical m / z value for BrN2OS is 380.06. The measured value is 381.1 [M+H]. + .

[0221] 5-Bromo-N-(3-(ethyl(4-fluorobenzyl)amino)propyl)thiophene-2-carboxamide (32a). 0.088 g (0.42 mmol) of 5-bromothiophene-2-carboxylic acid was dissolved in 1.7 mL of anhydrous DMF, and 0.177 g (0.47 mmol) of HATU was slowly added. After stirring for 15 min, 0.12 g (0.42 mmol) of 32b and 0.33 mL (1.91 mmol) of DIPEA were added to the mixture. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was diluted with DCM and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The mixture was purified by rapid chromatography (SiO₂; DCM / MeOH 1 - 4%) to give a white solid. The crude product was used without further purification. TLC-MS (ESI+): C 17 H 20 The theoretical m / z value for BrFN2OS is 398.05. The measured value is 399.1 [M+H]. + .

[0222] N-(3-aminopropyl)-5-phenylthiophene-2-carboxamide hydrochloride (34a). 0.6 g (1.66 mmol) of 33 was dissolved in 8.3 mL (33.3 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred overnight at room temperature. The resulting product was filtered off and dried under reduced pressure to give a pale yellow solid in 99% (0.486 g, 1.64 mmol). This product was used without further purification. 1 ¹H NMR (400 MHz, DMSO) δ 8.92 (t, J = 5.6 Hz, 1H), 8.10 (s, 3H), 7.88 (d, J = 3.9 Hz, 1H), 7.74 – 7.64 (m, 2H), 7.54 (d, J = 3.9 Hz, 1H), 7.44 (dd, J = 10.3, 4.7 Hz, 2H), 7.39 – 7.31 (m, 1H), 3.32 (q, J = 6.4 Hz, 2H), 2.89 – 2.77 (m, 2H), 1.84 (quintet, 2H). 13C NMR (101MHz, DMSO) δ 161.19, 147.33, 138.79, 133.12, 129.36, 129.24, 128.48, 125.64, 124.29, 39.52, 36.68, 36.10, 27.26. TLC-MS (ESI+): C 14 H 16 The theoretical m / z value for N2OS is 260.1. The measured value is 261.1 [M+H] + HPLC t R = 3.973 min.

[0223] 3-(3,4-dihydroisoquinoline-2(1H)-yl)propyl-1-amine dihydrochloride (35a). 0.61 g (2.1 mmol) of 35b was dissolved in 10.5 mL (42 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 1 h. The resulting product was filtered off by vacuum filtration and dried under reduced pressure. This product was used without further purification. TLC-MS (ESI+): C 12 H 18 The theoretical m / z value for N2 is 190.15. The measured value is 191.2 [M+H] + .

[0224] N-(3-(((3-aminopropyl)(methyl)amino)methyl)phenyl)-4-fluorobenzamide (36a). 0.506 g (1.22 mmol) of 36b was dissolved in 6.1 mL (24.4 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 3 h. After the reaction was complete, the mixture was diluted with DCM and washed with NH4Cl (3 x 20 mL), NaHCO3 (3 x 20 mL), and brine (3 x 20 mL). The combined organic phases were dried over Na2SO4, filtered, and evaporated to dryness to give a yellow oil. This crude product was used without further purification. TLC-MS (ESI+): C 18 H 22 The theoretical m / z value for FN3O is 315.17. The measured value is 316.2 [M+H]. + .

[0225] 1-(3-aminopropyl)-N-(tert-butyl)piperidine-4-carboxamide dihydrochloride (37a). 0.36 g (1.27 mmol) of 37b was dissolved in 6.3 mL (25.3 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 3 h. The HCl salt of the product formed was filtered off by suction filtration and dried under reduced pressure to give a white solid in 77% yield (0.306 g, 0.98 mmol). This product was used without further purification. 1 H NMR (400 MHz, DMSO) δ3.45 (d, J = 12.4 Hz, 2H), 3.16 (dd, J = 12.8, 7.7 Hz, 3H), 2.89 – 2.77 (m,4H), 2.39 – 2.29 (m, 1H), 2.11 – 1.78 (m, 8H), 1.25 (s, 2H), 1.22 (d, J = 4.8Hz, 9H). 13 C NMR (101 MHz, DMSO) δ 172.3, 52.9, 51.1, 49.9, 48.7, 36.3, 28.5, 25.7, 23.5, 21.4.

[0226] (1-Benzylpiperidin-3-yl)methylamine hydrochloride (39a). 0.165 g (0.54 mmol) of 39b was dissolved in 2.71 mL (10.8 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred overnight at room temperature. The resulting product was filtered off and dried under reduced pressure to give a white solid in 99% (0.163 g, 0.53 mmol). This product was used without further purification. 1 H NMR (400 MHz, DMSO) δ 8.55 – 8.01 (m, 3H), 7.76 –7.59 (m, 2H), 7.53 – 7.17 (m, 3H), 4.45 – 4.09 (m, 2H), 3.49 – 3.23 (m, 2H),3.11 – 2.57 (m, 4H), 2.46 – 2.28 (m, 1H), 2.02 – 1.69 (m, 3H), 1.19 – 1.00 (m, 1H). 13C NMR (101 MHz, DMSO) δ 131.72, 129.37, 129.32, 128.62, 59.39, 53.13, 51.15, 41.12, 31.82, 25.56, 21.24. TLC-MS (ESI+): C 13 H 20 The theoretical m / z value for N2 is 204.16. The measured value is 205.0 [M+H] + HPLC t R = 1.102 min.

[0227] N-(3-((1,4-diazacycloheptane-1-yl)methyl)phenyl)-4-fluorobenzamide hydrochloride (41a). 0.5 g (1.17 mmol) of 41b was dissolved in 5.9 mL (23.4 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 3 h. The HCl salt of the product formed was filtered off by suction filtration and dried under reduced pressure to give a white solid in 92% (0.392 g, 1.08 mmol). This product was used without further purification. 1 H NMR (400 MHz, DMSO) δ 10.53 (s, 1H), 9.91 (s, J = 37.5 Hz, 1H), 9.62 (s, 1H), 8.12 – 8.06 (m, 2H), 8.03 (s, 1H), 7.81 – 7.73 (m, J = 8.1 Hz, 1H), 7.52 – 7.34 (m, 4H), 3.75 – 3.56 (m, J = 25.4, 14.7 Hz, 4H), 3.48 – 3.10 (m, 4H), 2.54 (s, 2H), 2.30 – 2.09 (m, 2H). 13 C NMR (101 MHz, DMSO) δ 165.38, 164.51, 162.91, 139.39,131.06, 130.46, 130.23, 129.16, 126.66, 123.38, 121.64, 115.47, 66.35, 59.14,52.39, 48.83, 43.72, 40.43, 20.34. TLC-MS (ESI+): C 19 H 22 The theoretical m / z value for FN3O is 327.17. The measured value is 328.2 [M+H]. + .

[0228] (3-Nitrophenyl)methylamine dihydrochloride (42a). 1.13 g (4 mmol) of 42b was dissolved in 20 mL of EtOH, and 0.83 mL (20 mmol) of hydrazine hydrate (80% aqueous solution) was slowly added. The mixture was heated to 60 °C and stirred overnight. After the starting material was completely consumed, the precipitate formed was filtered off. The filtrate was concentrated and redissolved in DCM. A 1,4-dioxane solution of HCl was added to the suspension, and the solid formed was filtered off and washed several times with EtOAc. The product was dried under reduced pressure to give a brown solid. 1 H NMR (400 MHz, DMSO) δ 8.76 (s, 3H), 8.46 (t, J = 1.8Hz, 1H), 8.23 ​​(d, J = 8.2, 1.5 Hz, 1H), 8.01 (d, J = 7.7 Hz, 1H), 7.71 (t, J= 8.0 Hz, 1H), 4.18 (q, J = 5.8 Hz, 2H). 13 C NMR (101 MHz, DMSO) δ 147.65, 136.33, 135.99, 130.05, 124.01, 123.25, 41.26.

[0229] N-(3-Aminobenzyl)-5-phenylthiophene-2-carboxamide (43a). 0.224 g (0.66 mmol) of compound 42 was dissolved in 4 mL of ethanol (EtOH), and the solution was cooled to 0 °C. Under a nitrogen atmosphere, 0.04 g of 10% palladium on carbon (Pd / C) was added to the solution. The reaction flask was repeatedly evacuated and purged with hydrogen (H2), followed by bubbling of H2 in the suspension for 15 min. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction flask was purged with nitrogen, and the crude reaction mixture was filtered through a short diatomaceous earth pad. The reaction vessel and diatomaceous earth were washed with ethanol. The filtrates were combined and concentrated under reduced pressure. The residue was dried overnight under high vacuum to give a brown solid. The crude product was used directly without further purification. TLC-MS (ESI+): C 18 H 16 The theoretical m / z value for N2OS is 308.10. The measured value is 309.1 [M+H]. + .

[0230] 4-(3-aminopropyl)piperazine-1-carboxylic acid tert-butyl ester (46a). 4.5 g (12.2 mmol) of 46b was dissolved in 61 mL of EtOH, and 2.52 mL (61 mmol) of hydrazine hydrate (80% aqueous solution) was slowly added. The mixture was heated to 60 °C and stirred overnight. After the starting material was completely consumed, the precipitate formed was filtered off. The filtrate was concentrated and redissolved in DCM. The solution was washed with water (3 x 30 mL) and brine (3 x 30 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness to give a yellow oily substance in 77% yield (2.286 g, 9.4 mmol). This product was used without further purification. 1 ¹H NMR (400 MHz, CDCl₃) δ 3.45 – 3.37 (m, 4H), 2.73 (t, J = 6.8 Hz, 2H), 2.45 – 2.30 (m, 6H), 1.62 (quintet, 2H), 1.44 (s, 9H), 1.38 (s, J = 13.9 Hz, 2H). 13 C NMR (101 MHz, CDCl3) δ 154.9, 79.7, 56.6, 53.2, 40.8, 30.6, 28.5.

[0231] N-Benzylpiperidine-4-carboxamide hydrochloride (48a). 0.64 g (2 mmol) of 48b was dissolved in 10 mL (40 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 3 h. The HCl salt of the product formed was filtered off by suction filtration and dried under reduced pressure to give a white solid in 52% yield (0.266 g, 1.04 mmol). This product was used without further purification. 1 H NMR (400 MHz, DMSO) δ 8.89 (d, J = 7.3 Hz, 1H), 8.55 (t, J = 5.9 Hz, 1H), 7.37 – 7.17 (m, 5H), 4.25 (d, J = 5.9 Hz, 2H), 3.21 (t,J = 19.1 Hz, 2H), 2.93 – 2.74 (m, 2H), 2.50 – 2.44 (m, J = 7.5, 3.4 Hz, 1H), 1.97 – 1.67 (m, 4H). 13C NMR (101 MHz, DMSO) δ 173.1, 139.6, 128.3, 127.1, 126.8, 42.3, 41.9, 25.2.

[0232] (3-(benzyl(methyl)amino)propyl)carbamate tert-butyl ester (1b). 0.97 mL (7.56 mmol) of N-methyl-1-phenylmethylamine, 1.8 g (7.56 mmol) of (3-bromopropyl)carbamate tert-butyl ester, 4.93 g (15.12 mmol) of cesium carbonate, and 0.627 g (3.78 mmol) of potassium iodide were added to a flask containing 30.2 mL of acetone. The mixture was stirred at room temperature for 72 h. After the reaction was complete, the precipitate formed was filtered off, the filtrate was concentrated, and redissolved in DCM. The solution was washed with water (3 x 30 mL) and brine (3 x 30 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO2; DCM / MeOH 0 - 5%) to obtain a yellow oily substance in 53% yield (1.12 g, 4.08 mmol). TLC-MS (ESI+): C 16 H 26 The theoretical m / z value for N₂O₂ is 278.20. The measured value is 301.2 [M+Na]. + .

[0233] (3-((4-methoxybenzyl)(methyl)amino)propyl)carbamate tert-butyl ester (18b). 0.31 mL (2.1 mmol) of 1-(4-methoxyphenyl)-N-methylmethylamine, 0.5 g (2.1 mmol) of (3-bromopropyl)carbamate tert-butyl ester, 1.37 g (4.2 mmol) of cesium carbonate, and 0.174 g (1.05 mmol) of potassium iodide were added to a flask containing 8.4 mL of acetone. The mixture was stirred at room temperature for 72 h. After the reaction was complete, the precipitate formed was filtered off, the filtrate was concentrated, and redissolved in DCM. The solution was washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO2; DCM / MeOH 1 - 3%) to obtain a yellow oily substance in 49% (0.315 g, 1.03 mmol). 1¹H NMR (400 MHz, CDCl₃) δ 7.25 – 7.15 (m, 2H), 6.90 – 6.79 (m, 2H), 5.58 (s, 1H), 3.79 (s, 3H), 3.41 (s, 2H), 3.18 (q, J = 11.6, 5.7 Hz, 2H), 2.43 (t, J = 6.3 Hz, 2H), 2.15 (s, 3H), 1.67 (quintet, J = 6.3 Hz, 2H), 1.44 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 158.82, 156.25,130.90, 130.29, 130.29, 113.77, 113.77, 78.85, 62.05, 55.97, 55.35, 42.03,40.18, 28.61, 28.61, 28.61, 26.57. TLC-MS (ESI+): C 17 H 28 The theoretical m / z value for N₂O₃ is 308.21. The measured value is 331.2 [M+Na]. + .

[0234] (3-((4-bromobenzyl)(methyl)amino)propyl)carbamate tert-butyl ester (19b). 0.42 mL (2.1 mmol) of 1-(4-bromophenyl)-N-methylmethylamine, 0.5 g (2.1 mmol) of (3-bromopropyl)carbamate tert-butyl ester, 1.37 g (4.2 mmol) of cesium carbonate, and 0.174 g (1.05 mmol) of potassium iodide were added to a flask containing 8.4 mL of acetone. The mixture was stirred at room temperature for 72 h. After the reaction was complete, the precipitate formed was filtered off, the filtrate was concentrated, and redissolved in DCM. The solution was washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO2; PE / EtOAc 10 - 40%) to obtain a yellow oily substance in 47% (0.354 g, 0.99 mmol). 1¹H NMR (400 MHz, CDCl₃) δ 7.46 – 7.38 (m, 2H), 7.22 – 7.16 (m, J = 8.4 Hz, 2H), 5.43 (s, 1H), 3.42 (s, 2H), 3.18 (q, J = 11.6, 5.7 Hz, 2H), 2.43 (t, J = 6.4 Hz, 2H), 2.15 (s, 3H), 1.67 (quintet, J = 6.4 Hz, 2H), 1.44 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 156.19, 138.11,131.52, 131.52, 130.74, 130.74, 120.96, 78.97, 62.05, 56.07, 42.12, 40.00,28.61, 28.61, 28.61, 26.73. TLC-MS (ESI+): C 16 H 25 The theoretical m / z value for BrN₂O₂ is 356.11 / 358.11. The measured value is 379.1 / 381.1 [M+Na] + .

[0235] (3-((4-fluorobenzyl)(methyl)amino)propyl)carbamate tert-butyl ester (20b). 0.37 g (2.65 mmol) of 20c, 0.63 g (2.65 mmol) of (3-bromopropyl)carbamate tert-butyl ester, 1.72 g (5.3 mmol) of cesium carbonate, and 0.22 g (1.32 mmol) of potassium iodide were added to a flask containing 10.6 mL of acetone. The mixture was stirred at room temperature for 72 h. After the reaction was complete, the precipitate formed was filtered off, the filtrate was concentrated, and redissolved in DCM. The solution was washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness to give a yellow oil. This crude product was used without further purification.

[0236] (3-((2-fluorobenzyl)(methyl)amino)propyl)carbamate tert-butyl ester (21b). 0.28 mL (2.1 mmol) of 1-(2-fluorophenyl)-N-methylmethylamine, 0.5 g (2.1 mmol) of (3-bromopropyl)carbamate tert-butyl ester, 1.37 g (4.2 mmol) of cesium carbonate, and 0.174 g (1.05 mmol) of potassium iodide were added to a flask containing 8.4 mL of acetone. The mixture was stirred at room temperature for 72 h. After the reaction was complete, the precipitate formed was filtered off, the filtrate was concentrated, and redissolved in DCM. The solution was washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO2; PE / EtOAc 10 - 40%) to obtain a yellow oily substance in 69% (0.429 g, 1.45 mmol). 1 ¹H NMR (400 MHz, CDCl₃) δ 7.41 – 7.32 (m, J = 7.5, 1.5 Hz, 1H), 7.25 – 7.18 (m, J = 7.3, 4.7, 2.0 Hz, 1H), 7.14 – 7.07 (m, J = 7.5, 1.0 Hz, 1H), 7.05 – 6.98 (m, 1H), 5.39 (s, 1H), 3.55 (s, 2H), 3.18 (q, J = 11.8, 5.8 Hz, 2H), 2.47 (t, J = 6.4 Hz, 2H), 2.20 (s, 3H), 1.67 (quintet, 2H), 1.44 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 161.56, 156.25,131.51, 128.91, 125.48, 124.05, 115.41, 78.88, 55.96, 54.91, 42.02, 39.98,28.59, 26.81. TLC-MS (ESI+): C 16 H 25 The theoretical m / z value for FN₂O₂ is 296.19. The measured value is 319.2 [M+Na]. + .

[0237] (3-((3-fluorobenzyl)(methyl)amino)propyl)carbamate tert-butyl ester (22b). 0.29 mL (2.1 mmol) of 1-(3-fluorophenyl)-N-methylmethylamine, 0.5 g (2.1 mmol) of (3-bromopropyl)carbamate tert-butyl ester, 1.37 g (4.2 mmol) of cesium carbonate, and 0.174 g (1.05 mmol) of potassium iodide were added to a flask containing 8.4 mL of acetone. The mixture was stirred at room temperature for 72 h. After the reaction was complete, the precipitate formed was filtered off, the filtrate was concentrated, and redissolved in DCM. The solution was washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO2; PE / EtOAc 10 - 40%) to obtain a yellow oily substance in 56% yield (0.336 g, 1.18 mmol). 1 ¹H NMR (400 MHz, CDCl₃) δ 7.30 – 7.22 (m, 1H), 7.08 (d, J = 7.6 Hz, 1H), 7.06 – 7.01 (m, 1H), 6.97 – 6.89 (m, J = 8.3, 2.0 Hz, 1H), 5.45 (s, 1H), 3.46 (s, 2H), 3.20 (q, J = 11.8, 5.8 Hz, 2H), 2.45 (t, J = 6.3 Hz, 2H), 2.17 (s, 3H), 1.67 (quintet, 2H), 1.44 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 163.01, 156.23, 141.80, 129.84, 124.54, 115.76, 114.08,79.01, 62.23, 56.19, 42.16, 40.04, 28.57, 26.74. TLC-MS (ESI+): C 16 H 25 The theoretical m / z value for FN₂O₂ is 296.19. The measured value is 319.2 [M+Na]. + .

[0238] (3-(benzyl(ethyl)amino)propyl)carbamate tert-butyl ester (23b). 0.31 mL (2.1 mmol) of N-benzylethylamine, 0.5 g (2.1 mmol) of (3-bromopropyl)carbamate tert-butyl ester, 1.37 g (4.2 mmol) of cesium carbonate, and 0.174 g (1.05 mmol) of potassium iodide were added to a flask containing 8.4 mL of acetone. The mixture was stirred at room temperature for 72 h. After the reaction was complete, the precipitate formed was filtered off, the filtrate was concentrated, and redissolved in DCM. The solution was washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO₂; DCM / MeOH 2 - 7%) to give a yellow oil in 46% (0.282 g, 0.97 mmol). TLC-MS (ESI+): C 17 H 28 The theoretical m / z value for N₂O₂ is 292.22. The measured value is 315.2 [M+Na]. + .

[0239] (3-(benzyl(isopropyl)amino)propyl)carbamate tert-butyl ester (24b). 0.35 mL (2.1 mmol) of N-benzylpropyl-2-amine, 0.5 g (2.1 mmol) of (3-bromopropyl)carbamate tert-butyl ester, 1.37 g (4.2 mmol) of cesium carbonate, and 0.35 g (2.1 mmol) of potassium iodide were added to a flask containing 8.4 mL of acetone. The mixture was stirred at room temperature for 72 h. After the reaction was complete, the mixture was diluted with EtOAc and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na2SO4, filtered, and evaporated to dryness. The mixture was purified by rapid chromatography (SiO2; PE / EtOAc 0 -30%). The crude product was used without further purification. TLC-MS (ESI+): C 18 H 30 The theoretical m / z value for N₂O₂ is 306.23. The measured value is 329.2 [M+Na]. + .

[0240] (3-(benzyl(cyclopropyl)amino)propyl)carbamate tert-butyl ester (25b). 0.31 mL (2.1 mmol) of N-benzylcyclopropylamine, 0.5 g (2.1 mmol) of (3-bromopropyl)carbamate tert-butyl ester, 1.37 g (4.2 mmol) of cesium carbonate, and 0.35 g (2.1 mmol) of potassium iodide were added to a flask containing 8.4 mL of acetone. The mixture was stirred at room temperature for 72 h. After the reaction was complete, the mixture was diluted with EtOAc and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The mixture was purified by rapid chromatography (SiO₂; DCM / MeOH 0 - 3%). The crude product was used without further purification. TLC-MS (ESI+): C 18 H 28 The theoretical m / z value for N₂O₂ is 304.22. The measured value is 327.2 [M+Na]. + .

[0241] (3-(benzyl(propyl)amino)propyl)carbamate tert-butyl ester (26b). 0.35 mL (2.1 mmol) of N-benzylpropyl-1-amine, 0.5 g (2.1 mmol) of (3-bromopropyl)carbamate tert-butyl ester, 1.37 g (2.1 mmol) of cesium carbonate, and 0.174 g (1.05 mmol) of potassium iodide were added to a flask containing 8.4 mL of acetone. The mixture was stirred at room temperature for 72 h. After the reaction was complete, the mixture was diluted with EtOAc and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness to give a yellow oil. This crude product was used without further purification. TLC-MS (ESI+): C 18 H 30 The theoretical m / z value for N₂O₂ is 306.23. The measured value is 307.2 [M+H]. + .

[0242] (3-(benzyl(butyl)amino)propyl)carbamate tert-butyl ester (27b). 0.38 mL (2.1 mmol) of N-benzylbut-1-amine, 0.5 g (2.1 mmol) of (3-bromopropyl)carbamate tert-butyl ester, 1.37 g (4.2 mmol) of cesium carbonate, and 0.35 g (2.1 mmol) of potassium iodide were added to a flask containing 8.4 mL of acetone. The mixture was stirred at room temperature for 72 h. After the reaction was complete, the mixture was diluted with EtOAc and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The mixture was purified by rapid chromatography (SiO₂; DCM / MeOH 0 -3%). The crude product was used without further purification. TLC-MS (ESI+): C 19 H 32 The theoretical m / z value for N₂O₂ is 320.25. The measured value is 343.2 [M+Na]. + .

[0243] (3-(benzyl(3-hydroxypropyl)amino)propyl)tert-butyl carbamate (28b). 0.32 mL (2 mmol) of 3-(benzylamino)prop-1-ol, 0.478 mg (2 mmol) of (3-bromopropyl)tert-butyl carbamate, 1.3 g (4 mmol) of cesium carbonate, and 0.333 g (2 mmol) of potassium iodide were added to a flask containing 8 mL of acetone. The mixture was stirred at room temperature for 16 h. After the reaction was complete, the mixture was diluted with EtOAc and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO₂; DCM / MeOH + 1% NH₃ 0 - 5%) to give a colorless oil in 60% (0.388 g, 1.2 mmol). 1 H NMR(400 MHz, CDCl3) δ 7.36 – 7.22 (m, 5H), 3.72 (t, 2H), 3.54 (s, 2H), 3.09 (q,J = 6.6 Hz, 2H), 2.63 (t, J = 5.2 Hz, 2H), 2.46 (t, J = 7.0 Hz, 2H), 1.77 –1.64 (m, 4H), 1.46 – 1.40 (m, 11H). 13C NMR (101 MHz, CDCl3) δ 156.12, 138.44,129.29, 128.61, 127.44, 63.58, 59.03, 53.60, 51.45, 29.40, 28.55, 28.50,28.33, 27.16. TLC-MS (ESI+): C 18 H 30 The theoretical m / z value for N₂O₃ is 322.23. The measured value is 345.0 [M+Na]. + HPLC t R = 3.595 min.

[0244] (3-(benzyl(phenethyl)amino)propyl)carbamate tert-butyl ester (29b). 0.2 mg (0.94 mmol) of 29c, 0.225 mg (0.94 mmol) of (3-bromopropyl)carbamate tert-butyl ester, 0.616 g (1.89 mmol) of cesium carbonate, and 0.157 g (0.94 mmol) of potassium iodide were added to a flask containing 4 mL of acetone. The mixture was stirred at room temperature for 16 h. After the reaction was complete, the mixture was diluted with EtOAc and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO₂; DCM / MeOH 0 - 4%) to give a yellow oil in 51% (0.178 g, 0.48 mmol). 1 H NMR (400 MHz, CDCl3)δ 7.34 – 7.00 (m, 11H), 3.56 (s, 2H), 2.81 – 2.56 (m, 4H), 2.49 (t, J = 6.4Hz, 2H), 1.65 – 1.51 (m, 2H), 1.46 – 1.36 (m, 11H). 13 C NMR (101 MHz, CDCl3) δ155.99, 140.44, 139.35, 128.88, 128.70, 128.31, 128.27, 126.98, 125.93,58.81, 55.49, 52.14, 39.60, 33.39, 28.49, 28.44, 26.69. TLC-MS (ESI+): C 23 H 32 The theoretical m / z value for N₂O₂ is 368.25. The measured value is 369.1 [M+H]. +HPLC t R = 5.924 min.

[0245] (3-(benzyl(3-phenylpropyl)amino)propyl)tert-butyl carbamate (30b). 0.224 mg (1 mmol) of 30c, 0.237 mg (1 mmol) of (3-bromopropyl) tert-butyl carbamate, 0.649 g (2 mmol) of cesium carbonate, and 0.165 g (1 mmol) of potassium iodide were added to a flask containing 4 mL of acetone. The mixture was stirred at room temperature for 16 h. After the reaction was complete, the mixture was diluted with EtOAc and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO₂; DCM / MeOH 0 - 4%) to give a yellow oil in 48% (0.184 g, 0.48 mmol). 1 ¹H NMR (400 MHz, CDCl₃) δ 7.36 – 7.30 (m, 4H), 7.28 – 7.12 (m, 7H), 3.54 (s, 2H), 3.16 (q, J = 6.1 Hz, 2H), 2.60 (t, J = 6.8 Hz, 2H), 2.53 – 2.40 (m, 4H), 1.86 – 1.78 (m, 2H), 1.63 (quintet, J = 6.3 Hz, 2H), 1.43 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 156.15,142.41, 139.58, 129.13, 128.72, 128.49, 128.43, 127.11, 125.84, 78.82, 59.02,53.55, 52.69, 40.03, 33.77, 28.94, 28.61, 28.56, 26.63. TLC-MS (ESI+): C 24 H 34 The theoretical m / z value for N₂O₂ is 382.26. The measured value is 383.1 [M+H]. + HPLC t R = 6.308 min.

[0246] N 1 -Ethyl-N 1-(4-fluorobenzyl)propane-1,3-diamine dihydrochloride (32b). 0.731 g (2.35 mmol) of 32c was dissolved in 11.8 mL (47.1 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 1 h. The resulting product was filtered off by suction filtration and dried under reduced pressure to give a white semi-solid in 55% yield (0.368 g, 1.3 mmol). This product was used without further purification. TLC-MS (ESI+): C 12 H 19 The theoretical m / z value for FN2 is 210.15. The measured value is 211.2 [M+H]. + .

[0247] (3-(3,4-dihydroisoquinoline-2(1H)-yl)propyl)tert-butyl carbamate (35b). 0.26 mL (2.1 mmol) of 1,2,3,4-tetrahydroisoquinoline, 0.5 g (2.1 mmol) of (3-bromopropyl) tert-butyl carbamate, 1.37 g (4.2 mmol) of cesium carbonate, and 0.35 g (2.1 mmol) of potassium iodide were added to a flask containing 8.4 mL of acetone. The mixture was stirred at room temperature for 72 h. After the reaction was complete, the mixture was diluted with EtOAc and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The mixture was purified by rapid chromatography (SiO₂; DCM / MeOH 0 - 3%). The crude product was used without further purification. TLC-MS (ESI+): C 17 H 26 The theoretical m / z value for N₂O is 290.2 for². The measured value is 313.2 [M+Na]. + .

[0248] (3-((3-(4-fluorobenzamido)benzyl)(methyl)amino)propyl)carbamate tert-butyl ester (36b). 0.4 g (1.65 mmol) of 36c was dissolved in 15 mL of THF and one drop of acetic acid was added. After stirring for 15 min, 0.3 mL (1.5 mmol) of (3-(methylamino)propyl)carbamate tert-butyl ester was added to the mixture and the mixture was stirred for 1 h. The mixture was cooled to 0 °C and 0.476 g (2.25 mmol) of sodium triacetoxyborohydride was added. After the reaction was complete, the mixture was diluted with EtOAc and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was used without further purification. TLC-MS (ESI+): C 23 H 30 The theoretical m / z value for FN3O3 is 415.23. The measured value is 438.2 [M+Na]. + .

[0249] (3-(4-(tert-butylcarbamoyl)piperidin-1-yl)propyl)tert-butyl carbamate (37b). 0.3 g (1.36 mmol) of 37c, 0.324 g (1.36 mmol) of (3-bromopropyl) tert-butyl carbamate, 1.328 g (4.08 mmol) of cesium carbonate, and 0.113 g (0.68 mmol) of potassium iodide were added to a flask containing 5.44 mL of acetone. The mixture was stirred at room temperature for 16 h. After the reaction was complete, the mixture was diluted with EtOAc and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness to give a yellow semi-solid in 80% yield (0.371 g, 1.09 mmol). 1 H NMR (400 MHz, CDCl3) δ 5.40 (s, 1H), 5.24 (s, 1H), 3.22– 3.11 (m, 2H), 3.02 – 2.88 (m, J = 11.7 Hz, 2H), 2.37 (t, J = 6.7 Hz, 2H), 1.98 – 1.77 (m, 5H), 1.73 – 1.57 (m, 4H), 1.43 (s, 9H), 1.32 (s, 9H). 13C NMR (101 MHz, CDCl3) δ 174.3, 156.2, 57.1, 53.4, 51.1, 44.3, 40.1, 29.3, 29.0, 28.6, 28.5, 26.7. TLC-MS (ESI+): C 18 H 35 The theoretical m / z value for N3O3 is 341.27. The measured value is 342.3 [M+H] + .

[0250] ((1-Benzylpiperidin-3-yl)methyl)tert-butyl carbamate (39b). 0.16 mL (1.54 mmol) of benzaldehyde was dissolved in 14 mL of THF, and one drop of acetic acid was added. After stirring for 15 min, 0.3 g (1.4 mmol) of (piperidin-3-ylmethyl)carbamate tert-butyl was added, and the mixture was stirred for 1 h. The mixture was cooled to 0 °C, and 0.445 g (2.1 mmol) of sodium triacetoxyborohydride was added. After the reaction was complete, the mixture was diluted with EtOAc and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO₂; DCM / MeOH 0 - 7%) to give a yellow oil in 39% (0.165 g, 0.54 mmol). 1 H NMR (400 MHz, CDCl3) δ 7.54 – 7.27 (m, 5H), 4.66 (s, 1H), 3.65 (s,2H), 3.33 – 2.74 (m, 5H), 2.24 – 1.58 (m, 6H), 1.42 (s, 9H). TLC-MS (ESI+): C 18 H 28 The theoretical m / z value for N₂O₂ is 304.22. The measured value is 327.1 [M+Na]. + .

[0251] 4-(3-(4-fluorobenzamido)benzyl)-1,4-diazacycloheptane-1-carboxylic acid tert-butyl ester (41b). 0.37 g (1.52 mmol) of 36c was dissolved in 14 mL of THF and one drop of acetic acid was added. After stirring for 15 min, 0.27 mL (1.38 mmol) of 1,4-diazacycloheptane-1-carboxylic acid tert-butyl ester was added, and the mixture was stirred for 1 h. The mixture was cooled to 0 °C and 0.44 g (2.07 mmol) of sodium triacetoxyborohydride was added. After the reaction was complete, the mixture was diluted with EtOAc and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO2; DCM / MeOH 1 - 3%) to obtain a yellow oily substance in 92% yield (0.544 g, 1.27 mmol). 1 H NMR (400 MHz, CDCl3) δ 8.30 (d, J = 61.0 Hz, 1H), 8.01– 7.88 (m, 2H), 7.85 – 7.65 (m, 2H), 7.33 (t, J = 7.8 Hz, 1H), 7.14 (t, J =8.5 Hz, 3H), 3.80 (d, J = 9.5 Hz, 2H), 3.67 – 3.52 (m, J = 20.1 Hz, 2H), 3.53– 3.41 (m, 2H), 2.92 – 2.71 (m, J = 16.0 Hz, 4H), 2.02 – 1.93 (m, J = 10.7,5.5 Hz, 2H), 1.45 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 166.32, 165.01, 163.81,155.81, 155.46, 138.68, 131.08, 129.78, 129.46, 125.68, 121.36, 120.26,115.99, 115.77, 80.00, 61.91, 56.13, 54.85, 54.13, 45.58, 44.87, 28.60. TLC-MS(ESI+):C 22 H 32 The theoretical m / z value for N4O2S is 427.23. The measured value is 450.2 [M+Na]. + HPLC t R = 5.309 min.

[0252] 2-(3-Nitrobenzyl)isoindoline-1,3-dione (42b). 1 g (4.63 mmol) of 1-(bromomethyl)-3-nitrobenzene was dissolved in 14 mL of DMF, and 0.943 g (5.09 mmol) of potassium phthalimide was added in small portions. The reaction mixture was stirred at room temperature for 2 h. Water was added to the mixture, and the resulting suspension was filtered. The solid was washed several times with water and dried under reduced pressure at 50°C to give a brown solid in 92% yield (1.202 g, 4.26 mmol). 1 H NMR(400 MHz, DMSO) δ 8.22 – 8.17 (m, 1H), 8.16 – 8.10 (m, 1H), 7.96 – 7.83 (m,4H), 7.79 (d, J = 7.7 Hz, 1H), 7.63 (t, J = 7.9 Hz, 1H), 4.92 (s, 2H). 13 C NMR (101 MHz, DMSO) δ 167.70, 147.88, 138.83, 134.60, 134.19, 131.61, 130.17, 123.31, 122.45, 122.34, 40.23. TLC-MS (ESI+): C 15 H 10 The theoretical m / z value for N₂O₄ is 282.06. The measured value is 305.1 [M+Na]. + HPLC t R = 7.573 min.

[0253] 4-(3-(1,3-dioxoisoindoline-2-yl)propyl)piperazine-1-carboxylic acid tert-butyl ester (46b). 3 g (11.2 mmol) of N-(3-bromopropyl)phthalimide and 2.08 g (11.2 mmol) of 1-Boc-piperazine were dissolved in 45 mL of DMF. 2.3 mL (13.4 mmol) of DIPEA was added, and the mixture was heated to 100 °C for 72 h. After the reaction was complete, the mixture was diluted with EtOAc and washed with water (3 x 30 mL) and brine (3 x 30 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness to give a brown solid. This crude product was used without further purification. TLC-MS (ESI+): C 20 H 27 The theoretical m / z value for N3O4 is 373.20. The measured value is 374.2 [M+H]. + .

[0254] 4-(benzylcarbamoyl)piperidin-1-carboxylic acid tert-butyl ester (48b). 0.314 g (1.37 mmol) of 37e was dissolved in 4 mL of DCM, and 0.573 g (1.51 mmol) of HATU was slowly added. After stirring for 15 min, 0.45 mL (4.11 mmol) of phenylmethylamine and 0.21 mL (1.51 mmol) of triethylamine were added to the mixture. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was diluted with DCM and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness to give a brown solid. TLC-MS (ESI+): C 18 H 26 The theoretical m / z value for N₂O₃ is 318.19. The measured value is 341.2 [M+Na]. + .

[0255] 1-(4-fluorophenyl)-N-methylmethylamine (20c). 1.32 mL (2.65 mmol) of 2M methylamine in THF, 0.33 mL (2.65 mmol) of 1-(bromomethyl)-4-fluorobenzene, 1.72 g (5.29 mmol) of cesium carbonate, and 0.22 g (1.32 mmol) of potassium iodide were added to a flask containing 11 mL of acetone. The mixture was stirred at room temperature for 72 h. After the reaction was complete, the precipitate formed was filtered off, the filtrate was concentrated, and redissolved in DCM. The solution was washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness to give a yellow oil. This crude product was used without further purification.

[0256] N-Benzyl-2-phenylethyl-1-amine (29c). 0.25 mL (2.48 mmol) of benzaldehyde was dissolved in 12 mL of LEtOH. After stirring for 15 min, 0.31 mL (2.48 mmol) of 2-phenylethyl-1-amine was added, and the mixture was stirred for 1 h. The mixture was cooled to 0 °C, and 0.311 g (4.95 mmol) of sodium cyanoborohydride was added. After the reaction was complete, the solvent was removed under reduced pressure, and the mixture was diluted with water. The mixture was extracted with DCM (3 x 10 mL), washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO₂; DCM / MeOH + 1% NH₃ 0 - 4%) to give a yellow oil in 38% (0.2 g, 0.94 mmol). 1 H NMR (400 MHz, CDCl3) δ 7.33 – 7.13 (m, 11H), 3.76 (s, 2H), 2.91 – 2.83 (m, 2H), 2.83 – 2.75 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 140.40, 140.16, 128.85, 128.58, 128.51, 128.20, 127.03, 126.27, 53.99, 50.67, 36.49. TLC-MS (ESI+): C 15 H 17 The theoretical value of N's m / z is 211.14. The measured value is 212.0 [M+H] + HPLC t R = 3.352min.

[0257] N-Benzyl-3-phenylprop-1-amine (30c). 0.23 mL (2.22 mmol) of benzaldehyde was dissolved in 11 mL of LEtOH. After stirring for 15 min, 0.32 mL (2.22 mmol) of 3-phenylprop-1-amine was added to the mixture, and the mixture was stirred for 1 h. The mixture was cooled to 0 °C and 0.279 g (4.44 mmol) of sodium cyanoborohydride was added. After the reaction was complete, the solvent was removed under reduced pressure, and the mixture was diluted with water. The mixture was extracted with DCM (3 x 10 mL), washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO2; DCM / MeOH + 1% NH3 0 - 5%) to obtain a yellow oily substance in 33% (0.166 g, 0.73 mmol). 1 H NMR (400 MHz, CDCl3) δ 7.32 – 7.04 (m, 10H), 3.72 (s, 2H), 2.71 – 2.56 (m, 4H), 1.86 – 1.66 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 142.31, 140.61, 128.52, 128.45, 128.26, 127.03, 125.88, 54.15, 49.06, 33.77, 31.86. TLC-MS (ESI+): C 16 H 19 The theoretical value of N's m / z is 225.15. The measured value is 225.9 [M+H] + HPLC t R = 4.126 min.

[0258] (3-(ethyl(4-fluorobenzyl)amino)propyl)tert-butyl carbamate (32c). 0.48 mL (3.15 mmol) of N-(4-fluorobenzyl)ethylamine, 0.75 g (3.15 mmol) of (3-bromopropyl)tert-butyl carbamate, 2.05 g (6.3 mmol) of cesium carbonate, and 0.26 g (1.57 mmol) of potassium iodide were added to a flask containing 12.6 mL of acetone. The mixture was stirred at room temperature for 72 h. After the reaction was complete, the mixture was diluted with EtOAc and washed with water (3 x 10 mL) and brine (3 x 10 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO2; PE / EtOAc 10 – 30%) to obtain a yellow oily substance in 39% (0.377 g, 1.21 mmol). 1 ¹H NMR (400 MHz, CDCl₃) δ 7.32 – 7.25 (m, J = 8.5, 5.7 Hz, 2H), 7.03 – 6.95 (m, 2H), 5.47 (s, 1H), 3.49 (s, J = 14.4 Hz, 2H), 3.15 (q, J = 11.6, 5.7 Hz, 2H), 2.51– 2.39 (m, 4H), 1.61 (quintet, J = 12.6, 6.4 Hz, 2H), 1.43 (s, 9H), 1.03 (t, 3H). TLC-MS (ESI+): C 17 H 27 The theoretical m / z value for FN₂O₂ is 310.41. The measured value is 311.2 [M+H]. + .

[0259] 4-Fluoro-N-(3-formylphenyl)benzamide (36c). 10% hydrochloric acid (16 mL) was added dropwise to a stirred solution of 1.4 g (4.91 mmol) of 36c in 27 mL of acetone. The mixture was stirred at room temperature for 30 minutes. The solution was cooled to 4°C and stirred overnight. After the reaction was complete, the mixture was treated with NaHCO3 solution and the acetone was evaporated under reduced pressure. The remaining solid was filtered by vacuum filtration and washed several times with water. The solid product was dried under reduced pressure to give a white solid in 68% (0.812 g, 3.34 mmol). 1H NMR (400 MHz, CDCl3) δ 9.98 (s, 1H), 8.29 (s, 1H), 8.12 (d, J = 1.6 Hz, 1H), 8.03 (td, J = 8.2, 2.3 Hz, 1H), 7.96 –7.88 (m, 2H), 7.64 (t, J = 8.8 Hz, 1H), 7.56 – 7.48 (m, 1H), 7.19 – 7.11 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 192.2, 166.5, 165.1, 164.0, 139.0, 137.2, 130.0, 129.8, 129.7, 126.3, 126.2, 120.9, 116.2, 116.0. TLC-MS (ESI+): C 14 H 10 The theoretical m / z value for FNO2 is 243.07. The measured value is 266.2 [M+Na] + .

[0260] N-(tert-butyl)piperidine-4-carboxamide hydrochloride (37c). 1.47 g (5.17 mmol) of 37d was dissolved in 26 mL (104 mmol) of 4M HCl in 1,4-dioxane. The mixture was stirred at room temperature for 3 h. The resulting product was filtered off and dried under reduced pressure to give a white solid in 89% (1.015 g, 4.6 mmol). This product was used without further purification.

[0261] N-(3-(1,3-dioxolane-2-yl)phenyl)-4-fluorobenzamide (36d). 0.9 g (6.42 mmol) of 4-fluorobenzoic acid was dissolved in 26 mL of anhydrous DMF, and 2.69 g (7.07 mmol) of HATU was slowly added. After stirring for 15 min, 1.06 g (6.42 mmol) of 3-(1,3-dioxolane-2-yl)aniline and 2.8 mL (16.06 mmol) of DIPEA were added to the mixture. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was diluted with DCM and washed with water (3 x 20 mL) and brine (3 x 20 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness. The crude product was purified by rapid chromatography (SiO2; PE / EtOAc 0 - 30%) to obtain a white solid in 76% yield (1.4 g, 4.88 mmol). 1H NMR (400 MHz, DMSO) δ 10.32 (s, 1H), 8.08 – 8.02 (m, 2H), 7.87 (t, J = 1.7 Hz, 1H), 7.81 – 7.77 (m, J = 12.2, 4.0, 3.0 Hz, 1H), 7.39 –7.34 (m, J = 12.4, 5.2 Hz, 3H), 7.17 (d, J = 7.6 Hz, 1H), 5.73 (s, 1H), 4.09– 3.91 (m, 4H). 13 C NMR (101 MHz, DMSO) δ 164.5, 139.1, 138.7, 130.5, 130.4, 128.5, 122.0, 121.0, 118.5, 115.5, 115.3, 102.7, 64.8. TLC-MS (ESI+): C 16 H 14 The theoretical m / z value for FNO3 is 287.10. The measured value is 310.1 [M+Na] + .

[0262] 4-(tert-butylcarbamoyl)piperidine-1-carboxylic acid tert-butyl ester (37d). 1.23 g (5.36 mmol) of 37d was dissolved in 16 mL of DCM, and 2.24 g (5.9 mmol) of HATU was slowly added. After stirring for 15 min, 1.7 mL (16.1 mmol) of tert-butylamine and 0.82 mL (5.9 mmol) of triethylamine were added to the mixture. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was diluted with DCM and washed with water (3 x 20 mL) and brine (3 x 20 mL). The combined organic phases were dried over Na₂SO₄, filtered, and evaporated to dryness to give a white solid in 97% yield (1.48 g, 5.2 mmol). 1 H NMR (400 MHz, DMSO) δ 7.35 (s, 1H), 3.93 (d, J = 12.1 Hz, 2H), 2.77– 2.61 (m, J = 9.2 Hz, 2H), 2.23 (tt, J = 11.5, 3.6 Hz, 1H), 1.62 – 1.53 (m,J = 12.8, 2.2 Hz, 2H), 1.39 (s, 9H), 1.37 – 1.30 (m, 2H), 1.22 (s, 9H). 13C NMR (101 MHz, DMSO) δ 173.6, 153.8, 78.5, 49.7, 42.1, 38.2, 28.5, 28.3, 28.1. TLC-MS (ESI+): C 15 H 28 The theoretical m / z value for N₂O₃ is 284.21. The measured value is 307.2 [M+Na]. + .

[0263] 1-(tert-Butoxycarbonyl)piperidine-4-carboxylic acid (37e). 1 g (3.9 mmol) of 1-(tert-butyl) 4-ethylpiperidine-1,4-dicarboxylic acid was stirred with 29 mL (29.15 mmol) of 1M NaOH aqueous solution at room temperature for 16 h. After the reaction was complete, the mixture was acidified, and the precipitate was collected by vacuum filtration to give a white solid in 82% yield (0.731 g, 3.19 mmol). 1 H NMR (400 MHz, DMSO) δ 12.25 (s, 1H), 3.83 (d, J = 13.1 Hz, 2H), 2.81 (s, 2H), 2.39 (tt, J = 11.0, 3.8 Hz, 1H), 1.77 (dd, J = 13.3, 3.2 Hz,2H), 1.44 – 1.31 (m, 11H). 13 C NMR (101 MHz, DMSO) δ 175.7, 153.9, 78.6, 42.7, 40.1, 28.1, 27.8. TLC-MS (ESI-): C 11 H 19 The theoretical m / z value for NO4 is 229.13. The measured value is 228.1 [MH]. - .

Claims

1. A compound of any one of the general formulas (Ia) to (Ig): Or its stereoisomers, enantiomers, tautomers, prodrugs, and / or pharmaceutical salts. in X 1 Selected from substituted or unsubstituted cyclic groups, substituted or unsubstituted heterocyclic groups, substituted or unsubstituted fused cyclic groups, and substituted or unsubstituted fused heterocyclic groups. X 2 Selected from -SCH2-, -CH2CH2- and bonds, X 3 Selected from carbonyl, thiocarbonyl, and methylene. Y 1 Selected from H and methyl, Y 2 Selected from substituted or unsubstituted phenyl, substituted or unsubstituted benzyl, tert-butyl, and tert-butoxycarbonyl. Y 3 Selected from H, substituted or unsubstituted C1-C5 alkyl and C3-C5 cycloalkyl groups, Y 4 and Y 5 Is it O, or Y? 4 Selected from H and methyl, Y 5 Selected from substituted or unsubstituted benzyl, substituted or unsubstituted benzoyl, and straight-chain or branched C1-C5 alkyl carboxylic acid ester groups. A and B are independently selected from substituted or unsubstituted heterocyclic groups and substituted or unsubstituted fused heterocyclic groups. C is selected from substituted or unsubstituted cyclic groups, and n is 0-2.

2. The compound according to claim 1, wherein the compound has formula (Ia) or (Ib), wherein X 1 It is a substituted or unsubstituted heterocyclic group, preferably a substituted or unsubstituted thiophene-2-yl, more preferably a (substituted or unsubstituted phenyl)thiophene-2-yl. X 2 It is a key. X 3 It is a carbonyl group. Y 1 It's H. Y 2 It is an unsubstituted benzyl group. Y 3 Selected from straight-chain or branched C1-C4 alkyl, C2-C3 alkylphenyl and C3-C4 cycloalkyl, preferably methyl, and A is selected from substituted or unsubstituted 1-piperidinyl, substituted or unsubstituted 1-(2,3,4-trihydro)quinoline and substituted or unsubstituted 1-(2,3-dihydro)indole, and n is 1.

3. The compound according to any one of the preceding claims, wherein the compound has any one of general formulas (IIa) to (IIc): in R 1 yes , , , , , , , , , , , , , , , or , R 2 yes , , , , , , , , , , , or , R 3 yes , , , , , , , , , , , or ,and R 4 It is H or halogen, preferably F.

4. The compound according to any one of the preceding claims, wherein the compound has the general formula (IId) or (IIe): in R 5 yes , , , or .

5. The compound according to any one of the preceding claims, wherein the compound has 0.5 µM or less, preferably 0.1 µM or less, a β-repressor protein that recruits EC. 50 And / or more than 71%, preferably more than 90%, of P-selectin expression is reduced.

6. A pharmaceutical composition comprising a compound according to any one of claims 1 to 5, or a pharmaceutical salt, prodrug, biologically active metabolite, solvate, or stereoisomer thereof.

7. The pharmaceutical composition according to claim 6, wherein the pharmaceutical composition further comprises an inert carrier and / or one or more other therapeutic agents.

8. The compound according to any one of claims 1 to 5, or its pharmaceutical salt, prodrug, biologically active metabolite, solvate or stereoisomer, or the composition according to claim 6 or 7, for the treatment of a disease or condition.

9. The compound or composition according to claim 8, wherein the disease is mediated by ACKR3.

10. The compound or composition according to claim 8 or 9, wherein the disease or condition is selected from the group consisting of: pain, painful functional disorders or conditions of the subject, cancer, inflammatory or autoimmune diseases and conditions, conditions of excessive or abnormal angiogenesis (including wound healing and HIV infection), stem cell differentiation and mobilization disorders, brain and neuronal dysfunction (including Alzheimer's disease, multiple sclerosis and demyelinating diseases), renal dysfunction, renal insufficiency, preeclampsia and obesity.

11. The compound or composition according to any one of claims 8 to 10, wherein the disease is a cardiovascular disease or a platelet disorder, preferably the cardiovascular disease is selected from coronary artery disease, such as angina pectoris and heart attack, stroke, heart failure, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, arrhythmia, congenital heart disease, valvular heart disease, carditis, aortic aneurysm, peripheral artery disease, thromboembolic disease, venous thrombosis, and atherosclerosis, and wherein the platelet disorder is selected from thrombocytosis, thrombocytopenia, and thrombocytopenia, such as immune thrombocytopenia and thrombotic thrombocytopenic purpura, Behr-Soe syndrome, Grantzmann platelet atony, Hay-Putsch syndrome, Jacobson syndrome, Loehr's syndrome, platelet release and storage pool defects, thrombocytopenia with radius loss (TAR) syndrome, and thrombotic thrombocytopenic purpura (TTP).

12. A method for treating a disease, wherein ACKR3-mediated processes such as activation or inhibition, regulation, and modulation are beneficial in humans or warm-blooded animals or mammals requiring such treatment, the method comprising administering to the human or warm-blooded animal or mammal a therapeutically effective amount of a compound according to any one of claims 1 to 5, or a pharmaceutical salt, prodrug, biologically active metabolite, solvate, or stereoisomer thereof, or a composition according to claim 6 or 7.

13. The method of claim 12, wherein the disease is a cardiovascular disease or a platelet disorder.

14. The method of claim 13, wherein the cardiovascular disease is selected from coronary artery disease, such as angina pectoris and heart attack, stroke, heart failure, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, arrhythmia, congenital heart disease, valvular heart disease, carditis, aortic aneurysm, peripheral artery disease, thromboembolic disease, venous thrombosis, and atherosclerosis, and wherein the platelet disorder is selected from thrombocytosis, thrombocytopenia, and thrombocytopenia, such as immune thrombocytopenia and thrombotic thrombocytopenic purpura, Behr-Soe syndrome, Grantzmann platelet atony, Hay-Putsch syndrome, Jacobson syndrome, Loehr's syndrome, platelet release and storage pool defects, thrombocytopenia with radius loss (TAR) syndrome, and thrombotic thrombocytopenic purpura (TTP).

15. The method of claim 12, wherein the disease is selected from the group consisting of: painful functional disorders or conditions, acute or chronic pain, cancer (including breast cancer, brain cancer, lymphoma), fibrosis (including myocardial fibrosis), inflammatory or autoimmune diseases and conditions, conditions of excessive or abnormal angiogenesis (including wound healing), stem cell differentiation and mobilization disorders, brain and neuronal dysfunction (including Alzheimer's disease, multiple sclerosis, and demyelinating diseases), renal dysfunction, renal insufficiency, preeclampsia, and obesity.

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