The present invention relates to new tetrahydroquinoline (THQ) compounds and new methods for synthetizing such compounds.

The present invention relates to the applications thereof, in particular in therapeutic treatment.

Technological background

THQ synthesis is usually performed using the multicomponent Povarov reaction in cycloaddition [4+2] or sequential Mannich/Friedel-Craft reactions. In addition, the implementation of chiral catalysts such as Jacobsen's urea, phosphoric acid described by Zhu and Masson and, more recently, the silylated prolinol/POCl3/oxidation sequential reactions reported by Chan have been effective in obtaining 2,3,4-substituted THQs covering different configurations enantioselectively.

48-93% yield 87-98% ee 2) Friedel-Craft reaction (for R = EWG, POCI3 is required) H

3) Extraction up to 66-72% yield up to 99% ee

4) Dess-Martin periodinane dr > 95:5

However, access to the cis-2,3 and trans-3,4 configuration of enantioselectively 2,3,4-substituted THQs has not yet been obtained.

RVX208 ( quinazoline structure below) is the first molecule having a BD2 selectivity of BRD4 described in the literature and has been able to suppress the inflammatory response specifically for interleukin-6 (IL-6).

However, concerning THQ derivatives, PCT applications WO 201638120 A1 and WO 2014140076 A1 to Glaxo Smith Klime (GSK) describe a class of THQ being BET inhibitors.

However, THQ derivatives described by GSK are 2,3,4 substituted in configurations trans- 2,3 and trans- 3,4, because of the limitation of the existing synthesis routes which are not able to enantioselectively provide 2,3,4-THQ in the cis- 2,3 and trans- 3,4 configuration.

Goals of the invention

The present invention aims to solve the technical problem of providing a THQ substituted at least in 2,3,4 positions in another diastereomeric configuration and enantiomerically purethan the trans- 2,3 and trans- 3,4 configuration.

The present invention aims to solve the technical problem of providing a THQ substituted at least in 2,3,4 positions in the cis- 2,3 and trans- 3,4 configuration. The present invention aims to solve the technical problem of providing a process for preparing such THQ derivatives.

The present invention aims to solve the technical problem of providing a process preparing an enantiomeric mixture wherein one enantiomer of the THQ substituted at least in 2,3,4 positions in the cis- 2,3 and trans- 3,4 configuration predominates, and preferably, one enantiomer predominates to the extent of about 90% or greater, and preferably about 98% or greater (% in mol/mol of the enantiomer to the mixture of enantiomers).

Preferably, the present invention aims to solve the technical problem of providing a process easily implemented.

Preferably, the present invention aims to solve the technical problem of providing a process which could be implemented at the industrial scale.

Description of the invention

The present invention provides on one hand an innovative method of preparation of compounds according to the present invention, having a specific 2,3,4-THQ in the cis- 2,3 and trans- 3,4 configuration.

The present invention relates to compounds having the following structure: wherein R, R’, R _A , Nu, R1 , R2, R3, and R4 are each independently an atom or a chemical group of atoms.

In the present invention, the term compound relates to the compounds according to the recited structure or a salt thereof.

Advantageously, the compound of the invention is an enantiomer substantially free of other enantiomers or an enantiomeric mixture wherein one enantiomer of the compound according to the invention predominates. One enantiomer predominates to the extent of about 90% or greater, and preferably about 98% or greater (% in mol/mol of the enantiomer to the mixture of enantiomers).

In particular, the method according to the present invention allows the very specific configuration of the 2,3,4-THQ in the cis- 2,3 and trans- 3,4 configuration. In the prior art, such configuration is not obtainable in particular selectively and in enantiomerically pure form, and even less in a one pot reaction. Also, the method according to the present invention provides in a one pot reaction a 2,3,4-THQ in the trans- 2,3 and trans- 3,4 configuration. In the prior art, such configuration is not obtainable in a one pot reaction. Accordingly the present invention is very advantageous over the prior art. The cis-2,3 and trans-3,4 configuration of THQs is found to be essential for inhibitory activity on bromodomains and anti-inflammatory cellular effects. When the molecules are of trans- 2,3 and trans-3,4 configuration, no activity is observed on the inhibition of bromodomains (table XX), nor on cellular anti-inflammatory assays.

The present invention relates to compounds having the following structure: wherein R represents an alkyl group, wherein R1 , R2, R3, R4, R7 and R8 are each independently an atom or a chemical group of atoms, wherein -Nu represents a group of atoms, preferably selected from the group consisting of -N ₃ , -OR ₅ , -SR ₅ , -SeR ₅ and wherein -N is linked to the heterocycle and wherein the covalent bonds between N and R5, and N and R6 represent single or double bonds with R5 or R6, and wherein R5 and R6 represent each independently an atom or a chemical group of atoms and may form together a heterocycle, optionally substituted, or -Nu together with R form an heterocyle, for example thereby forming a hexahydropyrroloquinoline.

In particular, the compounds of the invention are enantiomers of structure A, A’, I, G

II or II’. Also, the present invention provides on the other hand compounds of structure A, A’, I, G II or IG according to the invention and structures falling within these definitions, because such compounds are good candidates for therapeutic treatments, for example for treating a disease involving an inflammatory disorder or a cancer. This is why the present invention focuses on these specific compounds.

In one preferred embodiment, R represents an alkyl group, typically a C1 -C6 alkyl, for example a methyl, ethyl or propyl. In an embodiment, R is CH ₃ (methyl).

In one preferred embodiment, R1 , R2 and R4 are hydrogen atoms.

In one preferred embodiment, R’ represents an alkyl group or -(CH ₂ )-R8 or-C(0)Rc, wherein Rc is a group of atoms, typically an alkyl, typically a C1 -C6 alkyl, for example a methyl, ethyl or propyl, wherein R8 is as defined in the present invention.

In an embodiment R8 is a group of atoms, typically an alkyl, typically a C1 -C6 alkyl, for example a methyl, ethyl or propyl. In an embodiment, R8 is CH ₃ (methyl).

In one preferred embodiment, R _A is H, an alkyl, -C(0)R7, -S(0) ₂ R7, -C(0)0R7,

In one preferred embodiment, R _A is H.

In one preferred embodiment, R _A is -C(0)R7.

In one preferred embodiment, R5 and R6 are each independently selected from the group consisting of H, C(0)0R ⁹ , an aryl or heteroaryl ring optionally substituted, wherein R9 is an alkyl group.

In one preferred embodiment, R5 and R6 form together an heteroaryl of the following structure: , wherein Ra and Rb are each independently an atom or a chemical group of atoms, preferably selected from the group consisting of H, a halogen atom (Br, Cl, F), an alkyl group optionally comprising one or more heteroatoms (O, N, S), an optionally substituted aryl or heteroaryl, for example an optionally substituted phenyl ring, and wherein Ra and Rb may form together a cycle or heterocycle, optionally substituted.

In one preferred embodiment, the compound according to the invention is of the following structure: In one preferred embodiment, the compound according to the invention is

In one preferred embodiment, Nu is N3. Advantageously, in such an embodiment wherein Nu is N ₃ , enable reactions of the click chemistry type to provide different compounds. Click Chemistry is a term describing reactions that are high yielding, wide in scope, creating only byproducts that can be removed without chromatography, and being stereospecific, simple to perform, and can be conducted in easily removable or benign solvents. Click chemistry is based on Azide-Alkyne Cycloaddition.

An example of click chemistry without the use of copper catalyst based compounds is: or the corresponding cis- 2,3 and trans- 3,4 enantiomer thereof. In one preferred embodiment,

Such compound may be for example obtained by reacting a compound of structure G or II’ with ^HC ^= Ra

In one embodiment, a compound according to the invention has the following structure:

or a corresponding cis- 2,3 and trans- 3,4 enantiomer thereof: or the corresponding cis- 2,3 and trans- 3,4 enantiomer thereof:

PROTAC

\ Spacer link wherein the spacer link is a covalent link comprising several atoms In one preferred embodiment, a compound according to the present invention has the following structure: corresponding cis- 2,3 and trans-

3,4 enantiomer thereof:

wherein the spacer link is a covalent link comprising several atoms. A skilled person knows many spacer links. The spacer link is functional to bind to or inhibit two targets, one target linking to one of the groups on each side of the spacer link. Such targets are for example BD1 and BD2.

An example of spacer link is -(CH ₂ 0)n-, wherein n is an integer, for example ranging from 2 to 10, typically n is 2, 3, 4, 5 or 6..

An example of spacer link is -CH ₂ -(OC ₂ H ₄ )n-OCH ₂ -, wherein n is an integer, for example ranging from 1 to 10, typically n is 2, 3, 4, 5 or 6. Also, in an embodiment, the compound of the present invention has a PROTAC function. Recently, PROTAC (Proteolysis-Targeting Chimeras) degraders have emerged as promising therapeutic strategy through utilization of the cells own protein destruction machinery to selectively degrade essential tumor drivers. A number of BET protein degraders (BBD) have been identified. In an embodiment, the compound of the present invention comprises one or more bromodomain and extra terminal domain (BET) proteolysis targeting chimera (PROTAC) (BET-PROTAC). In an embodiment a compound of the present invention has the following structure: PROTAC

\ Spacer link (VI) or the corresponding cis- 2,3 and trans- 3,4

PROTAC-

\ Spacer link enantiomer thereof: wherein the spacer link is a covalent link comprising several atoms, and PROTAC is a chemical group functioning as a PROTAC. A proteolysis targeting chimera (PROTAC) is a heterobifunctional small molecule composed of two active domains and a linker capable of removing specific unwanted proteins (the spacer link in the present invention). Rather than acting as a conventional enzyme inhibitor, a PROTAC works by inducing selective intracellular proteolysis. PROTACs consist of two covalently linked protein-binding molecules: one capable of engaging an E3 ubiquitin ligase, and another that binds to a target protein meant for degradation.

A skilled person knows many spacer links and PROTAC.

An example of spacer link is -(CH ₂ 0)n-, wherein n is an integer, for example ranging from 2 to 10, typically n is 2, 3, 4, 5 or 6.

An example of spacer link is -CH ₂ -(OC ₂ H ₄ )n-OCH ₂ -, wherein n is an integer, for example ranging from 1 to 10, typically n is 2, 3, 4, 5 or 6.

An example of such compounds are: , nd trans- 3,4 enantiomers thereof.

In another embodiment, compounds of the invention are of the following structure the corresponding cis- 2,3 and trans- 3,4 enantiomer thereof: wherein Rb, independently at each occurrence, has the same meaning as R1 , R2, R3, R4 and n represents the number of occurrence of Rb on the aromatic group.

In one embodiment of structure (VII), n is 1 and Rb is -NC(0)-(CH ₂ ) _m (C(0)NH0H, wherein m is an integer, for example ranging from 1 to 20, typically m is 2, 3, 4, 5, 6, 7, 8, 9 or 10. An example of such compound is: the corresponding cis- 2,3 and trans- 3,4 enantiomer thereof.

In one embodiment, R is an alkyl, preferably in C1 -C5, for example -C(CH ₂ )=CH ₂ , an alkoxy, for example an allyloxy or ^~ CH ₂ — CH (p _ro p-2-yn-1-yloxy), an aryl, preferably a phenyl group, and for example an alkyl-phenyl substituted on the phenyl group by a silane group, for example -CH _2. phenyl-0-Si-trialkyl.

In one preferred embodiment, in the compound according to the invention, in particular of structure (G) R and R8 are identical.

In one embodiment, R3 is H, OCH ₃ , O-alkyl, preferably 0-Ci-C ₅ alkyl, an ester, for example -C(0)0-alkyl, preferably C(0)0Me.

In an embodiment, R7 is an alkyl, typically a C1 to C5 alkyl.

In one embodiment, R7 is -CHsor -CH ₂ CH3. Preferably, R7 is -CH3.

In one embodiment, Nu is -S-aryl, for example -S-phenyl, -Se-aryl, for example - Se-phenyl,-NH-aryl, for example -NH-phenyl, OH, NH ₂ , -S-alkyl-SH.

In one embodiment, Nu is an isopropoxycarbonyl-amino group.

In one embodiment, Nu is an optionally substituted pyrimidin-2-yl-amino group.

In one embodiment, Nu is an optionally substituted 1 H-1 ,2,3-triazol-1-yl group.

In one embodiment, Nu is an optionally substituted (phenyl)-1 H-1 ,2,3-triazol-1-yl group.

In one embodiment, Nu is an optionally substituted 4-cyclohexyl-1 H-1 ,2,3-triazol-1- yl group.

In one embodiment, wherein Nu is a substituted 1 H-1 ,2,3-triazol-1 -yl group said triazole is substituted by a group comprising a polyethylene glycol group. Example of such groups are: 2-(prop-2-yn-1 -yloxy)ethoxy)methyl, oxybis(ethane-2,1- diyl))bis(oxy))bis(methylene), 2,5,8, 11 -tetraoxatetradec-13-yn-1-yl, 2,5,8, 11 ,14- pentaoxaheptadec-16-yn-1 -yl, 2,5,8, 11 ,14,17-hexaoxaicos-19-yn-1 -yl.

In one embodiment, -Nu and R form together a 5-member heterocycle, typically forming with the THQ ring a hexahydropyrroloquinoline. Such 5-member heterocycle can be substituted, for example by one or more alkyl groups and/or aryl groups, or -C(0)-alkyl, for example -C(0)-C ₂ H ₅ . Examples of substituted hexahydropyrroloquinoline are given in M. B. Haarr, M. O Sydnes, Molecules, 2021 , 26, 341 . In one embodiment, Nu is a link forming a dimeric THQ. Dimeric THQ are for example of the following structure: wherein

« linker » represents a group of atoms, preferably as defined for Nu groups, wherein at least one atom (preferably terminal atom) is further linked to the resulting part of the dimer.

In one embodiment, a compound according to the present invention is selected from the group consisting of:





or the corresponding cis- 2,3 and trans- 3,4 enantiomer thereof.

In one embodiment, the compound or the corresponding cis- 2,3 and trans- 3,4 enantiomer thereof.

In one embodiment, a compound of the present invention is a pure enantiomer. Pure” refers to a compound comprising less than 5%, preferably less than 4%, preferably less than 3%, preferably less than 2%, preferably less than 1%, preferably less than 0.5%, preferably less than 0.1%, even preferably less than 0.01%, of the other enantiomers.

The present invention relates to a method for providing tetrahydroquinoline (THQ) compounds as described according to the present invention.

More particularly, the present invention relates to a method for selective production of tetrahydroquinoline, said method comprising reacting reactive compounds in the presence of (i) L-Proline and/or D-Proline and/or (S) silyl prolinol and/or (R) silyl prolinol and (ii) a Lewis acid to provide a tetrahydroquinoline compound.

The present invention relates in particular to a new route to prepare asymmetric THQ according to an enantioselective way.

Preferably, the present invention is a one-pot method to provide tetrahydroquinoline compounds, in particular a compound of structure I or II.

In one preferred embodiment, the method according to the invention is implementing reactive compounds, which are easily commercially available.

Advantageously, a Lewis acid providing the aza-o-orthoquinone methide thereby allowing the possibility of grafting the Nu nucleophile.

In one preferred embodiment, the method according to the invention is implementing cheap catalysts, such as proline and BF ₃ .OEt2. To access to another configuration ( trans - 2,3 and trans- 3,4 configuration), silyl prolinol can be used

Advantageously, the present invention provides THQ substituted in positions 2,3,4 in the configuration cis- 2,3 and trans- 3,4 (with proline), and also trans- 2,3 and trans- 3,4 (with silyl prolinol) in a one pot reaction.

In one embodiment, the method according to the invention comprises the reaction with L-proline in association with a Lewis acid.

In one embodiment, the method according to the invention comprises the reaction with D-proline in association with a Lewis acid.

In one embodiment, the method according to the invention comprises the reaction with (S)-silyl prolinol in association with a Lewis acid.

In one embodiment, the method according to the invention comprises the reaction with (R)-silyl prolinol in association with a Lewis acid.

Typically, the method according to the present invention involves a Mannich/Friedel Craft reaction, preferably according to the Chan route. The classical Chan route implements a chiral silyl prolinol for the first Mannich reaction and then POCI ₃ for the Friedel Craft reaction. However, such classical route is unable to form the reactive intermediate azo- ortho-quinone methide provided by the present invention. In particular, the method of the present invention implements advantageously a Barbas-Mannich reaction in the presence of proline as catalyst starting from an imine, an aniline derivative and an aldehyde to yield a Mannich product in the cis-2,3 configuration.

A preferred aniline derivative has the following structure: wherein R1 , R2, R3 and R4 have the definition as mentioned in the present description.

Preferably, a Proline catalyst is implemented in this reaction to provides said Mannich product in the cis-2,3 configuration.

Preferably, in the presence of said Lewis acid based catalyst the very reactive aza- orthoquinone methide can be formed and react by an external nucleophile attack. Advantageously, according to the present invention the synthesis (the full sequence of reactions) is performed in one-pot, preferably by trapping (or scavenging) the aza- orthoquinone methide formed in situ by a nucleophilic compound.

Examples of Lewis acid based catalyst are the following compounds:

1 ) Lewis acid :

Lanthanide trifluoromethanesulfonates (ex : Yb(OTf) ₃ )

- Transition metals (ex : Y(OTf) ₃ )

Post-transition metal (ex : Zn(OTf) ₃ )

2) Organic Lewis acid :

- Carbocation (trityl), for example described by Naidu, Veluru Ramesh, Shengjun Ni, et Johan Franzen. « The Carbocation: A Forgotten Lewis Acid Catalyst ». ChemCatChem 7, n° 13 (3 juillet 2015): 1896-1905

(https://doi.Org/10.1002/cctc.201500225).

In one embodiment, said Lewis acid based catalyst is BF ₃ .OEt2.

In one embodiment, the method according to the invention comprises the following sequence in one-pot: wherein Rc is a group of atoms, typically an alkyl group, for example an ethyl group. The corresponding cis- 2,3 and trans- 3,4 enantiomer thereof may be obtained with the same reaction except reacting with D-proline instead of L-proline.

In one embodiment, R1 , R2 and R4 are hydrogen atoms.

In one embodiment, the method according to the invention comprises the following sequence in one-pot, wherein equivalents, temperature and solvents are given for illustrative purpose:

(50 mol%) 1 h

The corresponding cis- 2,3 and trans- 3,4 enantiomer thereof may be obtained with the same reaction except reacting with D-proline instead of L-proline. In one embodiment, the method according to the invention comprises the following sequence in one-pot, wherein equivalents, temperature and solvents are given for illustrative purpose: New multicomponent “one pot” general reaction

The corresponding cis- 2,3 and trans- 3,4 enantiomer thereof may be obtained with the same reaction except reacting with D-proline instead of L-proline.

New multicomponent "one pot" general reaction

The corresponding cis- 2,3 and trans- 3,4 enantiomer thereof may be obtained with the same reaction except reacting with D-proline instead of L-proline.

In one embodiment, the method according to the invention comprises the following sequence in one-pot, wherein equivalents, temperature and solvents are given for illustrative purpose: one pot general reaction wherein Rc is a group of atoms, typically an alkyl group, for example an ethyl group. one pot example of reaction wherein (4) and (5) represent respectively the alkyl chain length (4 or 5 carbon atoms). In one embodiment, said Lewis acid is BF ₃ OEt2 _.

In the present invention, the term carbon atom chain refers to a chain comprising in the present invention the term “alkyl group” refers to a linear, branched or cyclic alkyl group, optionally substituted and/or optionally containing one or more heteroatoms in the alkyl chain. According to the invention, the term “alkyl group” refers to a linear or branched alkyl group, optionally substituted. According to an embodiment, the alkyl group may contain one or more heteroatoms in the alkyl chain. Preferably the "alkyl group" is a linear or branched, substituted or unsubstituted, C1 -C10 alkyl group, such as C1 -C4 or C1 -C6, in particular a methyl, ethyl group, propyl group, preferably methyl. A cyclic alkyl group is a cycloalkyl group. The term "cycloalkyl group" means a saturated or partially unsaturated, monocyclic, fused bicyclic or bridged polycyclic ring assembly containing the number of ring atoms indicated. This includes substituted or unsubstituted cylcoalkyl groups. For example, cycloalkyl group may be C3-C10 alkyl group, such as C5 or C6, in particular a cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl group etc. Cycloalkyl groups include heterocycle, by replacing one or more carbon atoms of the cycloalkyl group by one or more nitrogen, oxygen, or sulphur atoms an alkyl group containing one or more oxygen (heteroatom) is a cycloalkyl group. Examples of suitable alkoxy groups include methoxy, ethoxy, n-propoxy, l-propoxy, n-butoxy, s-butoxy, t-butoxy. The term "aryl group" means a substituted or unsubstituted monocylic or fused bicycic aromatic ring assembly containing six to ten ring carbon atoms. For example, aryl may be phenyl or naphthyl, preferably phenyl. The term "heteroaryl group" is as defined for aryl group where one or more of the ring members is a heteroatom. For example heteroaryl groups includes pyridyl, indolyl, indazolyl, quinoxalinyl, quinolinyl, benzofuranyl, benzopyranyl, benzothiopyranyl, benzo[l,3]dioxole, imidazolyl, benzo-imidazolyl, pyrimidinyl, furanyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, thienyl, etc. Aryl and heteroaryl also include akylaryl and alkylheteroaryl groups, i.e. an aryl or heteroaryl group linked by an alkyl part to the rest of the molecule. Also, Aryl and heteroaryl include aralkyl and heteroaralkyl groups, i.e. aryl and heteroaryl substituted by one or more alkyl groups.

Advantageously, the present invention provides a very simple method to provide the compounds of the invention.

Advantageously, the method according to the present invention is reproducible and do not needs to implement hazardous reacting conditions.

Advantageously, the method according to the present invention is ecofriendly, i.e. economical in the number of synthesis steps (less time, less purification, less solvents consumed), use of a natural catalyst, i.e. proline, catalytic synthesis (thereby economical in the quantity of reagents).

BET proteins (Bromodomain and ExtraTerminal Domain) are regulators of epigenetic mechanisms involved in gene expression. The BET proteins (BRD2, BRD3, BRD4 and BRDT) have two bromodomains (BD), namely BD1 and BD2, which recognize acetylated lysins in histones. These C-terminal regions of histones are involved in the recruitment of molecular complexes essential for the activation and regulation of transcription. Inhibitors that prevent bromodomains from interacting with these regions are recent targets of choice in therapeutics and referred to as BET inhibitors (iBETs). The compounds of the present invention, in particular compounds of formula (I) or (II), are BET inhibitors.

Advantageously, a compound of the invention, in particular a compound of formula (I) or (II) is for use as a selective BET inhibitor.

Advantageously, the compounds of the invention, in particular compounds of formula (I) or (II), have shown efficacy in inhibiting BRD.

In one embodiment, the compounds of the present invention, in particular compounds of formula (I) or (II), are BRD4 inhibitors.

Preferred compounds according to the invention, in particular compounds of formula (I) or (II), present a selectivity for BD2 toward BD1 . Advantageously, the compounds of the invention, in particular compounds of formula (I) or (II), present a very low cell toxicity.

Advantageously, the compounds of the present invention, in particular compounds of formula (I) or (II), are anti-inflammatory compounds.

Advantageously, the compounds of the invention, in particular compounds of formula (I) or (II), are effective in inhibiting the expression of IL-6 and TNF-a.

Advantageously, the compounds of the present invention, in particular compounds of formula (I) or (II), are good candidates for anti-cancer compounds.

In one embodiment, the compounds of the present invention are used in the treatment of an inflammation, preferably involving expression of IL-6 and/or TNF-a, or BET- dependent cancers: hematological malignancies (leukemia, lymphoma, multiple myeloma), solid tumors (prostate cancer, breast, lung,...) or inflammation-induced cancers (for example hepatocellular carcinoma).

According to WO 2016/038120, Bromodomain inhibitors are believed to be useful in the treatment of a variety of diseases or conditions related to systemic or tissue inflammation, inflammatory responses to infection or hypoxia, cellular activation and proliferation, lipid metabolism, fibrosis and in the prevention and treatment of viral infections.

Bromodomain inhibitors may be useful in the treatment of a wide variety of acute or chronic autoimmune and/or inflammatory conditions such as rheumatoid arthritis, osteoarthritis, acute gout, psoriasis, systemic lupus erythematosus, multiple sclerosis, inflammatory bowel disease (Crohn's disease and Ulcerative colitis), asthma, chronic obstructive airways disease, pneumonitis, myocarditis, pericarditis, myositis, eczema, dermatitis (including atopic dermatitis), alopecia, vitiligo, bullous skin diseases, nephritis, vasculitis, hypercholesterolemia, atherosclerosis, Alzheimer's disease, depression, Sjogren's syndrome, sialoadenitis, central retinal vein occlusion, branched retinal vein occlusion, Irvine-Gass syndrome (post cataract and post-surgical), retinitis pigmentosa, pars planitis, birdshot retinochoroidopathy, epiretinal membrane, cystic macular edema, parafoveal telengiectasis, tractional maculopathies, vitreomacular traction syndromes, retinal detachment, neuroretinitis, idiopathic macular edema, retinitis, dry eye (keratoconjunctivitis Sicca), vernal keratoconjunctivitis, atopic keratoconjunctivitis, uveitis (such as anterior uveitis, pan uveitis, posterior uveitis, uveitis-associated macular edema), scleritis, diabetic retinopathy, diabetic macula edema, age-related macular dystrophy, hepatitis, pancreatitis, primary biliary cirrhosis, sclerosing cholangitis, Addison's disease, hypophysitis, thyroiditis, type I diabetes, type II diabetes, giant cell arteritis, nephritis including lupus nephritis, vasculitis with organ involvement such as glomerulonephritis, vasculitis including giant cell arteritis, Wegener's granulomatosis, Polyarteritis nodosa, Behcet's disease, Kawasaki disease, Takayasu's Arteritis, pyoderma gangrenosum, vasculitis with organ involvement and acute rejection of transplanted organs.

Bromodomain inhibitors may be useful in the treatment of diseases or conditions which involve inflammatory responses to infections with bacteria, viruses, fungi, parasites or their toxins, such as sepsis, acute sepsis, sepsis syndrome, septic shock, endotoxaemia, systemic inflammatory response syndrome (SIRS), multi-organ dysfunction syndrome, toxic shock syndrome, acute lung injury, ARDS (adult respiratory distress syndrome), acute renal failure, fulminant hepatitis, burns, acute pancreatitis, post-surgical syndromes, sarcoidosis, Herxheimer reactions, encephalitis, myelitis, meningitis, malaria and SIRS associated with viral infections such as influenza, herpes zoster, herpes simplex and coronavirus. In one embodiment the disease or condition which involves an inflammatory response to an infection with bacteria, a virus, fungi, a parasite or their toxins is acute sepsis.

Bromodomain inhibitors may be useful in the treatment of viral infections.

Bromodomain inhibitors may be useful in the treatment of cancer, including hematological (such as leukaemia, lymphoma and multiple myeloma), epithelial including lung, breast and colon carcinomas, midline carcinomas, mesenchymal, hepatic, renal and neurological tumours.

In one embodiment the disease or condition for which a bromodomain inhibitor is indicated is selected from diseases associated with systemic inflammatory response syndrome, such as sepsis, burns, pancreatitis, major trauma, haemorrhage and ischaemia. In this embodiment the bromodomain inhibitor would be administered at the point of diagnosis to reduce the incidence of SIRS, the onset of shock, multi-organ dysfunction syndrome, which includes the onset of acute lung injury, ARDS, acute renal, hepatic, cardiac or gastro-intestinal injury and mortality. In another embodiment the bromodomain inhibitor would be administered prior to surgical or other procedures associated with a high risk of sepsis, haemorrhage, extensive tissue damage, SIRS or MODS (multiple organ dysfunction syndrome). In a particular embodiment the disease or condition for which a bromodomain inhibitor is indicated is sepsis, sepsis syndrome, septic shock and endotoxaemia. In another embodiment, the bromodomain inhibitor is indicated for the treatment of acute or chronic pancreatitis. In another embodiment the bromodomain is indicated for the treatment of burns.

As used herein the reference to the "treatment" of a particular disease or condition includes the prevention or prophylaxis of such a disease or condition.

The term "diseases or conditions for which a bromodomain inhibitor is indicated", is intended to include each of or all of the above diseases or conditions. While it is possible that for use in therapy, a compound of the present invention as well as pharmaceutically acceptable salts thereof may be administered as the raw chemical, it is common to present the active ingredient as a pharmaceutical composition.

In one embodiment, a compound of the present invention or a composition of the present invention is for use in a method for treating an inflammation or a cancer, preferably a cancer involving BET over-expression.

The present invention also relates to a composition, in particular a pharmaceutical composition, comprising a compound as described according to the present invention.

In a further aspect, the present invention relates to a pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers, diluents or excipients.

The composition of the present invention may conveniently be presented for use in the form of a pharmaceutical composition and thus pharmaceutical compositions comprising a combination as defined above together with a pharmaceutically acceptable diluent or carrier represent a further aspect of the invention.

The present invention also relates to a method for preparing a medicament or pharmaceutical composition comprising one or more compounds as defined in the present invention.

The present invention also relates to the use of one or more compounds as defined in the present invention in a method for preparing a medicament or pharmaceutical composition.

The present invention also relates to a method of therapeutic treatment of a human, said method comprising the administration of one or more compounds as defined in the present invention to said human in an effective amount, preferably in a medicament or pharmaceutical composition.

In the present invention, the terms “according to the invention” or “of the present invention” or the like refer to all and any embodiments, preferred and advantageous features alone or in any possible combination.

Other aims, characteristics and advantages of the invention will appear clearly to the person skilled in the art upon reading the explanatory description which makes reference to the Examples which are given simply as an illustration and which in no way limit the scope of the invention.

The Examples make up an integral part of the present invention, and any characteristic which appears novel with respect to any prior state of the art from the description taken in its entirety, including the Examples, makes up an integral part of the invention in its function and in its generality.

Thus, every example has a general scope. Furthermore, in the Examples, all percentages are given by weight, unless indicated otherwise, temperature is expressed in degrees Celsius unless indicated otherwise, and the pressure is atmospheric pressure, unless indicated otherwise.

On the figures

Figure 1 shows a graphic representing the evaluation of cytotoxicity after iBET/LPS treatment (on J774 mouse macrophages).

Figure 2 shows a graphic representing the evaluation of IL-6 cytokine expression after iBET/LPS treatment at 24h of different compounds.

Figure 3 shows a graphic representing the evaluation of IL-6 cytokine expression after iBET/LPS treatment at 48h of different compounds.

Figure 4 shows a graphic representing the evaluation of TNF-a cytokine expression after iBET/LPS treatment at 24h of different compounds.

Figure 5 shows a graphic representing the evaluation of TNF-a cytokine expression after iBET/LPS treatment at 48h of different compounds.

Figure 6 shows a graphic representing the HTFtF signal as a function of different inhibitors concentration (MLE-6-161 , MLE-6-162 are compounds of the invention whereas MLE-7-74, MLE-7-75, MLE-7-76 and JQ1 are compounds of the prior art).

Figure 7 concerns the evaluation of cytotoxicity and anti-inflammatory activity of BET inhibitor compounds. In this figure, the bubble plots represent anti-inflammatory activity (Al score) versus cytotoxicity (log IC ₅ o) for all compounds tested.

Figure 8 concerns the impact of “lead” BET inhibitors compounds on gene expression profiles. A. Volcano plot representing impact of iBET on gene expression profiles as indicated. B. Table and proportional Venn diagram representing number and overlap of differentially expressed (DE) genes for each iBET. C. Heat map representation of differentially expressed (DE) genes for each iBET. Subset on genes commonly downregulated for all iBET is circled. D. GSEA plots representing the biological pathway commonly downregulated. EXAMPLES

(2S,3S,4fl)-Ethyl-4-azido-6-methoxy-3-pentyl-1 ,2,3,4-tetrahydroquinoline-2- carboxylate (7)

To a solution of ethyl (E)-2-((4-methoxyphenyl)imino)acetate 2 (300.0 mg, 1.45 mmol) and freshly distilled heptanal (248.0 mg, 2.17 mmol) in anhydrous dioxane (10 ml.) was added L-proline (26.4 mg, 0.21 mmol) at rt. The reaction mixture was stirred for 48 h at the same temperature. After the reaction was complete, TMSN ₃ (216.0 mg, 1 .88 mmol) and BF _3. Et ₂ 0 (90 mI_, 0.73 mmol) were added and the mixture was stirred at rt for 1 h. The reaction was quenched with NaOH (2 N) until neutralization, extracted with EtOAc, dried by MgSC> ₄ , filtered off and concentrated by vacuum. After flash chromatography (silica gel, 5% EtOAc/cyclohexane), the product 7 was isolated as an oil (333.0 mg, 66.4%); R _f = 0.27 (10% EtOAc/cyclohexane); [a] _D ²⁰ = +126.6 (c = 1.03, CHCI ₃ ); IR v _max (thin film, CH ₂ CI ₂ ) 3393, 2954, 2927, 2091 , 1734, 1622, 1501 , 1463, 1224 cm ¹ ; UV : 208, 245, 325; ¹ H NMR (400 MHz, CDCIs) <5 ppm 0.84 (t, J = 6.9 Hz, 3H), 0.94-1.29 (m, 8H), 1.32 (t, J = 7.1 Hz, 3H), 2 .24 (m, 1 H), 3.76 (s, 3H), 4.19 (m, 1 H), 4.26 (m, 1 H), 4.29 (m, 1 H), 4.40 (d, J = 2.4 Hz, 1 H), 6.64 (d, J = 8.8 Hz, 1 H), 6.68 (d, J = 2.8 Hz, 1 H), 6.81 (dd, J = 8.8, 2.8 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) d ppm 14.1 (CH ₃ ), 14.4 (CH ₃ ), 22.6 (CH ₂ ), 26.4 (CH ₂ ), 27.0 (CH ₂ ), 31.8 (CH ₂ ), 38.6 (CH), 53.5 (CH), 56.0 (CH ₃ ), 61.4 (CH), 61.6 (CH ₂ ), 115.6 (CH), 115.8 (C), 116.3 (CH), 117.1 (CH), 136.8 (C), 151.9 (C), 172.5 (C); LRMS (ESI+) m/z (%) 347 (20) [/W+H] ⁺ , 304 (100); HRMS (ESI+) m/z calc for Ci ₈ H ₂₇ N ₄ 0 ₃ 347.2078, found 347.2077. (2S,3S,4fi)-Ethyl-6-methoxy-3-pentyl-4-(phenylthio)-1,2,3,4- tetrahydroquinoline-2-carboxylate (8)

From imine 2 (300.0 mg, 1.45 mmol), freshly distilled heptanal (248.0 mg, 2.17 mmol) and thiophenol (206.0 mg, 1.88 mmol) in dioxane (10 ml_), and using method A, the product was obtained after flash chromatography (silica gel, 5% EtOAc/ cyclohexane) to give 8 (329.0 mg, 55%); R _f = 0.17 (5% EtOAc/cyclohexane); [a] _D ²⁰ = +110 (c = 1 .08, CHCI ₃ ); IR V _max (thin film, CH ₂ CI ) 3393, 2900, 1731 , 1504, 1463, 1215, 1150 cm ¹ ; UV: 206, 223, 245, 328; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.76 (J = 7.3 Hz, 3H), 0.93-1.21 (m, 8H), 1.28 (t, J = 7.1 Hz, 3H), 2.30 (m, 1 H), 3.69 (s, 3H), 4.17-4.28 (m, 2H), 4.28 (d, J = 1.8 Hz, 1 H), 4.68 (d, J = 2.8 Hz, 1 H), 6.57 (d, J = 8.6 Hz, 1 H), 6.70 (dd, J = 8.6, 2.8 Hz, 1 H), 6.73 (d, J = 2.8 Hz, 1 H), 7.28-7.38 (m, 3H), 7.53 (dd, J= 8.1 , 1 .4 Hz, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 14.1 (CH ₃ ), 14.3 (CH ₃ ), 22.6 (CH ₂ ), 26.5 (CH ₂ ), 27.3 (CH ₂ ), 31.4 (CH ₂ ), 37.8 (CH), 50.5 (CH), 53.5 (CH), 55.9 (CH ₃ ), 61.3 (CH ₂ ), 115.9 (CH), 116.2 (2xCH), 118.0 (C), 127.9 (CH), 129.2 (2xCH), 133.4 (2xCH), 134.9 (C), 137.1 (C), 152.0 (C), 173.1 (C); LRMS (ESI+) m/z (%) 414 (20) [M+ H] ⁺ , 304 (100); HRMS (ESI+) m/z calc for C ₂₄ H ₃₂ N0 ₃ S 414.2097, found 414.2093.

(2S,3S,4fi)-Ethyl-6-methoxy-3-pentyl-4-(phenylselanyl)-1 ,2,3,4- tetrahydroquinoline-2-carboxylate (9)

From imine 2 (300.0 mg, 1.45 mmol), freshly distilled heptanal (248.0 mg, 2.17 mmol) and benzeneselenol (296.0 mg, 1 .88 mmol), and using method A, the product was obtained after flash chromatography (silica gel, 5% EtOAc/cyclohexane) to give 9 (220.0 mg, 33%); R _f = 0.23 (5% EtOAc/cyclohexane); [a] _D ²⁰ = +135 (c = 1 .01 , CHCI ₃ ); IR v _max (thin film, CH ₂ CI ₂ ) 3398, 2926, 1733, 1505, 1467, 1213, 1036 cm ¹ ; UV: 206, 226, 335; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.75 (J = 7.3 Hz, 3H), 0.86-1.18 (m, 8H), 1.29 (t, J = 7.1 Hz, 3H), 2.30 (m, 1 H), 3.70 (s, 3H), 4.17-4.30 (m, 2H), 4.52 (d, J = 1.6 Hz, 1 H), 4.79 (d, J = 2.8 Hz, 1 H), 6.54 (d, J = 8.7 Hz, 1 H), 6.67 (dd, J= 8.7, 2.9 Hz, 1 H), 6.72 (d, J = 2.8 Hz, 1 H), 7.29- 7.37 (m, 3H), 7.65 (m, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 14.1 (CH ₃ ), 14.4 (CH ₃ ), 22.6 (CH ₂ ), 26.5 (CH ₂ ), 27.4 (CH ₂ ), 31 .3 (CH ₂ ), 38.3 (CH), 46.2 (CH), 54.2 (CH), 55.9 (CH ₃ ), 61 .4 (CH ₂ ), 115.6 (CH), 155.9 (CH), 116.2 (CH), 118.9 (C), 128.3 (CH), 129.3 (2xCH), 130.3 (C), 135.8 (2xCH), 136.9 (C), 152.0 (C), 173.2 (C); LRMS (ESI+) m/z (%) 462 (15) [M+ H] ⁺ , 304 (100); HRMS (ESI+) m/zcalc. for C ₂₄ H ₃₂ N0 ₃ Se 462.1543, found 462.1542.

(2S,3/?,4fi)-Ethyl-4-((4-chlorophenyl)amino)-6-methoxy-3- pentyl-1,2,3,4- tetrahydroquinoline-2-carboxylate (10a)

From imine 2 (300.0 mg, 1.45 mmol), freshly distilled heptanal (248.0 mg, 2.17 mmol) and 4-chloroaniline (221.0 mg, 1.74 mmol), and using method A, the product was obtained after flash chromatography (silica gel neutralized by Et ₃ N, 5% EtOAc/cyclohexane) to give 10a (315.0 mg, 50.5%) and 10b (31.0 mg, 5%);

10a : R _f = 0.23 (10% EtOAc/cyclohexane); [a] _D ²⁰ = +80.2 (c = 1.03, CHCI ₃ ); IR v _max (thin film, CH ₂ CI ₂ ) 3393, 2954, 2924, 1725, 1595, 1501 , 1460, 1212, 1150 cm ¹ ; UV : 206, 258, 315; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.88 (t, J = 7.0 Hz, 3H), 1.09 (m, 1 H), 1.15- 1.35 (m, 7H), 1 .27 (t, J = 7.1 Hz, 3H), 2.41 (m, 1 H), 3.68 (s, 3H), 4.10 (bs, 1 H), 4.14-4.31 (m, 2H), 4.36 (bs, 1 H), 6.57-6.61 (m, 3H), 6.70 (d, J= 2.8 Hz, 1 H), 6.75 (dd, J= 8.8, 2.8 Hz, 1 H), 7.16 (d, J = 8.8 Hz, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) d ppm 14.2 (CH ₃ ), 14.4 (CH ₃ ), 22.7 (CH ₂ ), 26.9 (CH ₂ ), 27.3 (CH ₂ ), 32.2 (CH ₂ ), 37.9 (CH), 53.7 (CH), 54 .4 (CH), 55.9 (CH ₃ ), 61 .5 (CH ₂ ), 113.8 (2xCH), 115.6 (CH), 116.3 (CH), 116.6 (CH), 119.6 (C), 122.1 (C), 129.4 (2xCH), 136.8 (C), 145.1 (C), 152.3 (C), 173.3 (C); HRMS (ESI-) m/zcalc. for C ₂₄ H ₃ oN ₂ 0 ₃ CI 429.1950, found 429.1949. (2S,3/?,4S)-Ethyl-4-((4-chlorophenyl)amino)-6-methoxy-3-pent yl-1 ,2,3,4- tetrahydroquinoline-2-carboxylate (10b)

10b: R r = 0.20 (10% EtOAc/cyclohexane); [a] _D ²⁰ = +1.02 (c = 0.89, CHCI ₃ ); IR v _max (thin film, CH ₂ CI ₂ ) 3390, 2954, 2927, 2853, 2087, 1725, 1595, 1507, 1203, 1033 cm ¹ ; UV : 206, 258, 315; ¹ H NMR (400 MHz, CDCI ₃ ) d ppm 0.86 (t, J = 7.1 Hz, 3H), 0.91 (t, J = 6.8 Hz, 3H), 1 .24-1 .56 (m, 8H), 2.80 (m, 1 H), 3.71 (s, 3H), 3.76 (m, 1 H), 3.95 (m, 1 H), 4.09 (d, J = 4.2 Hz, 1 H), 4.19 (bs, 1 H), 6.45 (d, J = 8.8 Hz, 2H), 6.63 (d, J = 2.8 Hz, 1 H), 6.64 (d, J = 8.8 Hz, 1 H), 6.78 (dd, J = 8.8, 2.8 Hz, 1 H), 7.13 (d, J = 8.8 Hz, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 13.9 (CH ₃ ), 14.3 (CH ₃ ), 22.8 (CH ₂ ), 27.4 (CH ₂ ), 31.2 (CH ₂ ), 32.1 (CH ₂ ), 36.7 (CH), 54.0 (CH), 54.5 (CH), 56.0 (CH ₃ ), 61 .3 (CH ₂ ), 113.3 (2xCH), 115.5 (CH), 116.4 (CH), 116.6 (CH), 119.8 (C), 121.8 (C), 129.3 (2xCH), 136.6 (C), 145.1 (C), 152.5 (C), 175.6 (C); HRMS (ESI-) m/z calc for C ₂₄ H ₃ oN ₂ 0 ₃ CI 429.1950, found 429.1945. (2S,3S,4fl)-ethyl 4-{[(fi)-2-acetamido-3-methoxy-3-oxopropyl]thio}-6-methoxy-

3-pentyl-1,2,3,4-tetrahydroquinoline-2-carboxylate (11)

From imine 2 (300.0 mg, 1.45 mmol), freshly distilled heptanal (248.0 mg, 2.17 mmol) and methyl acetyl -/.-cystein ate (281.0 mg, 1.59 mmol), and using method A, the product was obtained after flash chromatography (silica gel, 40% EtOAc/cyclohexane) to give 11 (320.6 mg, 46%); R _f = 0.29 (50% EtOAc/cyclohexane); [a] _D ²⁰ = -11.6 (c = 1.19, CHCh); IR v _max (thin film, CH ₂ CI ) 3385, 2929, 1730, 1656, 1505, 1370, 1216, 1153 cm ¹ ; UV: 205, 246, 330; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.85 (t, J = 7.0 Hz, 3H), 1 .08 (m, 1 H), 1.14-1.29 (m, 7H), 1.34 (t, J = 7.1 Hz, 3H), 2.07 (s, 3H), 2.31 (m, 1 H), 2.94 (dd, J = 13.7, 6.3 Hz, 1 H), 3.23 (dd, J = 13.7, 4.5 Hz, 1 H), 3.75 (s, 3H), 3.79 (s, 3H), 3.93 (d, J = 1.6 Hz, 1 H), 4.22-4.37 (m, 2H), 4.50 (m, 1 H), 4.94 (ddd, J = 7.6, 6.3, 4.5 Hz, 1 H), 6.54 (d, J = 8.7 Hz, 1 H), 6.69 (dd, J= 8.7, 2.8 Hz, 1 H), 6.73 (d, J= 2.8 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 14.2 (CH ₃ ), 14.5 (CH ₃ ), 22.7 (CH ₂ ), 23.4 (CH ₃ ), 27.1 (CH ₂ ), 27.7 (CH ₂ ), 31.9 (CH ₂ ), 33.8 (CH ₂ ), 39.0 (CH), 46.2 (CH), 52.1 (CH ₃ ), 53.0 (CH), 53.7 (CH), 55.9 (CH ₃ ), 61 .6 (CH ₂ ), 115.9 (CH), 116.0 (CH), 116.1 (CH), 118.2 (C), 136.7 (C), 152.0 (C), 170.1 (C), 171.4 (C), 173.1 (C); LRMS (ESI+) m/z (%) 481 (1) [M+ H] ⁺ , 302 (100); HRMS (ESI+) m/z calc for C ₂₄ H ₃₇ N ₂ 0 ₆ S481 .2367, found 481 .2357.

(2/?,3/?,4S)-ethyl 4-{[(fl)-2-acetamido-3-methoxy-3-oxopropyl]thio}-6-methoxy- 3-pentyl-1 ,2,3,4-tetrahydroquinoline-2-carboxylate (12)

From 2 (300.0 mg, 1 .45 mmol), freshly distilled heptanal (248.0 mg, 2.17 mmol) and methyl acetyl-/.-cysteinate (281.0 mg, 1.59 mmol), and using the method A and D-proline as catalyst, the product was obtained after flash chromatography (silica gel, 40% EtOAc/cyclohexane) to give 12 (346.4 mg, 50%); R _f = 0.27 (50% EtOAc/cyclohexane); [a] _D ²⁰ = +123.8 (c = 1 .00, CHCIs); IR v _max (thin film, CH ₂ CI ) 3303, 2954, 2927, 2863, 1739, 1655, 1503, 1467, 1290, 1230 cnr ¹ ; UV: 206, 247, 333; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.85 (t, J = 7.0 Hz, 3H), 1.06 (m, 1 H), 1 .13-1 .30 (m, 7H), 1.33 (t, _* 7= 7.1 Hz, 3H), 2.05 (s, 3H), 2.30 (m, 1 H), 3.08 (dd, J = 13.9, 4.9 Hz, 1 H), 3.14 (dd, J = 13.9, 4.7 Hz, 1 H), 3.75 (s, 3H), 3.80 (s, 3H), 3.85 (d, J = 1.7 Hz, 1 H), 4.31 -4.36 (m, 2H), 4.51 (d, J = 2.9 Hz, 1 H), 4.95 (dt, J = 7.8, 4.8 Hz, 1 H), 6.54 (dd, J = 7.8, 1.2 Hz, 1 H), 6.67-6.70 (m, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 14.1 (CH ₃ ), 14.5 (CH ₃ ), 22.6 (CH ₂ ), 23.3 (CH ₃ ), 27.1 (CH ₂ ), 27.7 (CH ₂ ), 31.9 (CH ₂ ), 34.1 (CH ₂ ), 39.7 (CH), 47.0 (CH), 52.2 (CH ₃ ), 52.8 (CH), 53.6 (CH), 55.9 (CH ₃ ), 61.6 (CH ₂ ), 115.8 (CH), 116.0 (CH), 116.1 (CH), 118.2 (C), 136.7 (C) , 152.0 (C), 170.0 (C), 171.4 (C), 173.0 (C); HRMS (ESI+) m/z calc for C ₂₄ H ₃₇ N ₂ 0 ₆ S 481 .2367, found 481.2361. Ethyl-(2S,3S,4fi)-4-(((1 /?,2S,5fi)-2-isopropyl-5-methylcyclohexyl)oxy)-6- methoxy-3-pentyl-1,2,3,4-tetrahydroquinoline-2-carboxylate (13)

From imine 2 (300.0 mg, 1.45 mmol), freshly distilled heptanal (248.0 mg, 2.17 mmol) and (-)-menthol (295.0 mg, 1.88 mmol), and using method A, the product was obtained after flash chromatography (silica gel, 5% EtOAc/cyclohexane) to give 13 (217.0 mg, 32.6%); R _f = 0.29 (5% EtOAc/cyclohexane); [a] _D ²⁰ = +24.6 (c = 1.58, CHCI ₃ ); IR v _max (thin film, CH ₂ CI ₂ ) 3404, 2920, 1733, 1503, 1460, 1366, 1213, 1154, 1040 cm ¹ ; UV :226, 246; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.82-0.91 (m, 9H), 0.98 (d, J = 6.6 Hz, 3H), 1.02- 1.34 (m, 13H), 1.32 (t, J = 7.1 Hz, 3H), 1.42 (m, 1 H), 1.63-1.72 (m, 2H), 2.26-2.35 (m, 3H),

3.31 (td, J = 10.4, 4.1 Hz, 1 H), 4.17 (d, J = 1.9 Hz, 1 H), 4.23 (m, 1 H), 4.23-4.30 (m, 2H), 6.60 (m, 1 H), 6.71 -6.75 (m, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 14.2 (CH ₃ ), 14.4 (CH ₃ ), 16.0 (CH ₃ ), 21 .6 (CH ₃ ), 22.6 (CH ₃ , CH ₂ ), 23.1 (CH ₂ ), 25.2 (CH), 25.8 (CH ₂ ), 27.4 (CH ₂ ), 31 .9 (CH), 32.0 (CH ₂ ), 34.7 (CH ₂ ), 38.6 (CH), 41 .7 (CH ₂ ), 48.2 (CH), 53.7 (CH), 56.0 (CH ₃ ), 61 .3 (CH ₂ ), 73.5 (CH), 76.9 (CH), 115.9 (CH), 116.2 (CH), 116.8 (CH), 120.2 (C), 137.3 (C),

152.3 (C), 173.5 (C); HRMS (ESI+) m/z calc for C ₂₈ H ₄₆ N0 ₄ 460.3421 , found 460.3419.

Ethyl-(2/?,3/?,4S)-4-(((1 /?,2S,5fl)-2-isopropyl-5-methylcyclohexyl)oxy)-6- methoxy-3-pentyl-1,2,3,4-tetrahydroquinoline-2-carboxylate (14)

From imine 2 (300.0 mg, 1.45 mmol), freshly distilled heptanal (248.0 mg, 2.17 mmol) and (-)-menthol (295 mg, 1 .88 mmol), and using method A and D-proline as catalyst, the product was obtained after flash chromatography (silica gel, 5% EtOAc/cyclohexane) to give 14 (232.5 mg, 35%); R _f = 0.4 (5% EtOAc/cyclohexane); [a] _D ²⁰ = -108.6 (c = 0.89, CHCI ₃ ); IR V _max (thin film, CH ₂ CI ₂ ) 3402, 2930, 2089, 1505, 1463, 1250 cm ¹ ; UV: 225, 246; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.42 (d, J= 6.9 Hz, 3H), 0.80 (d, J= 7.1 Hz, 3H), 0.84 (t, J = 7.0 Hz, 3H), 0.96 (d, J = 6.6 Hz, 3H), 1.01 -1.32 (m, 13H), 1.33 (t, J = 7.1 Hz, 3H), 1.42 (m, 1 H), 1.54-1.68 (m, 2H), 2.10 (m, 1 H), 2.25 (m, 1 H), 2.33 (m, 1 H), 3.32 (td, J= 10.5, 4.0 Hz, 1 H), 3.74 (s, 3H), 4.20-4.32 (m, 2H), 4.29 (m, 1 H), 4.41 (m, 1 H), 6.58 (d, J = 8.7 Hz, 1 H), 6.65 (d, J = 2.8 Hz, 1 H), 6.74 (dd, J = 8.7, 2.8 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 14.2 (CH _S ), 14.5 (CH ₃ ), 15.5 (CH ₃ ), 21 .3 (CH ₃ ), 22.6 (CH), 22.7 (CH ₃ ), 23.2 (CH ₂ ), 25.4 (CH), 26.1 (CH ₂ ), 27.3 (CH ₂ ), 31 .9 (CH), 32.0 (CH ₂ ), 34.8 (CH), 38.8 (CH), 41 .6 (CH ₂ ), 48.9 (CH), 53.4 (CH), 56.1 (CH ₃ ), 61.3 (CH ₂ ), 73.8 (CH), 75.5 (CH), 115.7 (CH), 115.9 (CH), 117.0 (CH), 118.8 (C), 137.4 (C), 151.3 (C), 173.9 (C); HRMS (ESI+) m/z calc for C ₂₈ H ₄₆ N0 ₄ 460.3421 , found 460.3426.

(2S,3S,4fl)-Ethyl 4-hydroxy-6-methoxy-3-pentyl-1,2,3,4-tetrahydroquinoline-2- carboxylate (15)

From imine 2 (300.0 mg, 1.45 mmol), freshly distilled heptanal (248.0 mg, 2.17 mmol) and water (34.0 mg, 1.88 mmol), and using method A, the product was obtained after flash chromatography (silica gel, 10% to 15% EtOAc/cyclohexane) to give 15 (125.6 mg, 27%); R _f = 0.33 (20% EtOAc/cyclohexane); [a] _D ²⁰ = +33.3 (c = 1.00, CHC ); IR v _max (thin film, CH ₂ CI ₂ ) 3395, 2956, 2923, 2851 , 1730, 1503, 1461 , 1284, 1227, 1153, 1027 crrv ¹ ; UV: 208, 246, 322; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.84 (t, J = 6.9 Hz, 3H), 0.85-1.31 (m, 8H), 1 .32 (t, J = 7.1 Hz, 3H), 2.31 (m, 1 H), 4.22-4.32 (m, 2H), 4.25 (d, J = 3.2 Hz, 1 H), 4.52 (d, J = 2.4 Hz, 1 H), 6.62 (m, 1 H), 6.76-6.79 (m, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 14.1 (CH ₃ ), 14.4 (CH ₃ ), 22.6 (CH ₂ ), 26.0 (CH ₂ ), 27.2 (CH ₂ ), 31.9 (CH ₂ ), 40.5 (CH), 53.0 (CH), 55.9 (CH ₃ ), 61.4 (CH ₂ ), 69.6 (CH), 115.5 (CH), 116.1 (CH), 116.6 (CH), 121.1 (C), 136.8 (C), 152.1 (C), 173.4 (C); LRMS (ESI+) m/z (%) 322 (60) [M+ H] ⁺ , 304 (100); HRMS (ESI+) m/z calc for Ci ₈ H ₂₈ N0 ₄ 322.2013, found 322.2013. (2S,3S,4S)-Ethyl-4-azido-3-pentyl-1 ,2,3,4-tetrahydroquinoline-2-carboxylate

(16)

To a solution of aniline (108.0 mg, 1.16 mmol) and ethylglyoxylate (0.28 ml_, 1.39 mmol, 50% in toluene) in toluene was added MgSC>4 (87.0 mg, 0.72 mmol) at rt. The reaction mixture was stirred at 110°C for 1 h. After cooling rt, the reaction was filtered off and co evaporated with toluene two times. The mixture was diluted in dioxane (8 ml.) and freshly distilled heptanal (199.0 mg, 1.74 mmol) and L-proline (20.0 mg, 0.17 mmol) were added at rt. The reaction mixture was stirred at this temperature for 48 h. After the reaction was complete TMSN ₃ (213.0 mg, 1 .51 mmol) and BF _3. Et ₂ 0 (54 mI_, 0.58 mmol) were added at rt and the reaction was stirred at this temperature for 1 h30. The reaction was quenched by NaOH (2 N), extracted with EtOAc, dried by MgS0 ₄ , filtered off and concentrated in vacuum. The product was obtained after flash chromatography (silica gel, 5% EtOAc/cyclohexane) to give 16 (140.0 mg, 38%); R _f = 0.21 (5% EtOAc/cyclohexane); [a] _D ²⁰ = + 151.3 (c = 1.10, CHCIs); IR v _max (thin film, CH ₂ CI ) 3427, 2932, 2857, 2093, 1732, 1620, 1496, 1259, 1220 crrv ¹ ; UV : 225, 252; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.84 (t, J = 6.9 Hz, 3H), 1.02 (m, 1 H), 1.08-1 .29 (m, 7H), 1 .33 (t, J = 7.1 Hz, 1 H), 2.24 (m, 1 H), 4.21 -4.35 (m, 3H), 4.44 (d, J = 2.5 Hz, 1 H), 6.66 (d, J= 8.1 Hz, 1 H), 6.71 (td, = 7.4, 1.1 Hz, 1 H), 7.10 (dd, J= 7.4, 1 .1 Hz, 1 H), 7.16 (ddd, = 8.1 , 7.4, 1.5 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 14.1 (CH ₃ ), 14.4 (CH ₃ ), 22.6 (CH ₂ ), 26.4 (CH ₂ ), 27.0 (CH ₂ ), 31 .8 (CH ₂ ), 38.2 (CH), 53.2 (CH), 61 .2 (CH), 61.7 (CH ₂ ), 114.8 (C), 114.9 (CH), 117.4 (CH), 130.2 (CH), 131.2 (CH), 142.7 (C), 172.5 (C); HRMS (ESI+) m/z calc for Ci ₇ H ₂₄ 0 ₂ N ₄ Li 323.2054, found 323.2047.

(2S,3S,4fl)-Ethyl-4-azido-6-methoxy-3-methyl-1,2,3,4-tetr ahydroquinoline-2- carboxylate (17)

From imine 2 (450.0 mg, 2.17 mmol), recently purchased propioaldehyde (189.0 mg, 3.26 mmol) and TMSN ₃ (275.0 mg, 2.82 mmol), and using method A, the product was obtained after flash chromatography (silica gel, 4% EtOAc/cyclohexane) to give 17 as an oil (373.0 mg, 59.5%); R _f = 0.48 (7% EtOAc/cyclohexane); [a] _D ²⁰ = +126.1 (c = 1 .00, CHCI ₃ ); IR V _max (thin film, CH ₂ CI ₂ ) 3384, 2933, 2090, 1731 , 1501 , 1463, 1251 cm ¹ ; UV : 205, 249, 326; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.76 (d, J = 7.0 Hz, 3H), 1.32 (t, J = 7.1 Hz, 3H), 2.40 (m, 1 H), 3.76 (s, 3H), 4.18 (dd, J = 2.9, 2.3 Hz, 1 H), 4.24-4.34 (m, 3H), 6.65 (d, J = 8.8 Hz, 1 H), 6.68 (d, J= 2.8 Hz, 1 H), 6.81 (dd, J= 8.8, 2.9 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 12.0 (CH ₃ ), 14.4 (CH ₃ ), 33.3 (CH), 53.2 (CH), 56.0 (CH ₃ ), 61.7 (CH ₂ ), 63.4 (CH), 115,4 (C), 115.6 (CH), 116.4 (CH), 117.2 (CH), 136.4 (C), 151.9 (C), 172.4 (C); HRMS (ESI+) m/z calc for CI ₄ HI ₉ N ₄ 0 ₃ 291 .1452, found 291 .1447.

(2S,3S,4fl)-ferf-Butyl-4-azido-6-methoxy-3-methyl-1 ,2,3,4- tetrahydroquinoline-2-carboxylate (18)

From tert- butyl (£)-2-((4-methoxyphenyl)imino)acetate ¹ (341.0 mg, 1.45 mmol), recently purchased propioaldehyde (126.0 mg, 2.17 mmol) and TMSN ₃ (218.0 mg, 1.88 mmol), and using method A, the product was obtained after flash chromatography (silica gel, 5% to 10% EtOAc/cyclohexane) to give 18 (221.0 mg, 48%); R _f = 0.37 (5% EtOAc/cyclohexane); [a] _D ²⁰ = +145.2 (c = 1.11 , CHCI ₃ ); IR v _max (thin film, CH ₂ CI ) 3398, 2977, 2090, 1725, 1503, 1366, 1240, 1157, 1040 cm ¹ ; UV : 247; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.76 (d, J= 7.0 Hz, 3H), 1.51 (s, 9H), 2.35 (m, 1 H), 3.75 (s, 3H), 4.06 (d, = 3.1 Hz, 1 H), 4.22 (d, J = 2.4 Hz, 1 H), 6.63 (d, J = 8.8 Hz, 1 H), 6.67 (d, J = 2.8 Hz, 1 H), 6.80 (dd, J = 8.8, 2.8 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 11 .8 (CH ₃ ), 28.2 (3xCH ₃ ), 33.2 (CH), 53.4 (CH), 56.0 (CH ₃ ), 63.6 (CH), 82.3 (C), 115.3 (C), 115.6 (CH), 116.3 (CH), 117.3 (CH), 136.6 (C), 151.8 (C), 171.6 (C); LRMS (ESI+) m/z (%) 319 (10) [M+ H] ⁺ , 220 (100); HRMS (ESI+) m/z calc for Ci ₆ H ₂₃ N ₄ 0 ₃ 319.1770, found 319.1764. (2S,3S,4fl)-Ethyl-4-azido-3-(4-((ferf-butyldimethylsilyl)oxy )benzyl)-6-methoxy- 1 ,2,3,4-tetrahydroquinoline-2-carboxylate (19)

From imine 2 (300.0 mg, 1.45 mmol), 3-(4 -((tert- butyldimethylsilyl)oxy)phenyl)propanal ² (574.0 mg, 2.17 mmol) and TMSN ₃ (216.0 mg, 1.88 mmol), and using the method A, the product was obtained after flash chromatography (silica gel, 5% EtOAc/cyclohexane) to give 19 as a white solid (485.0 mg, 67.5%); R 0.38 (5% EtOAc/cyclohexane); [a] _D ²⁰ = +40.5 (c = 1 .00, CHCI ₃ ); IR v _max (thin film, CH ₂ CI ) 3406, 2956, 2934, 2854, 2094, 1737, 1605, 1509, 1466, 1257, 1229 cm ¹ ; UV : 205, 249, 326; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.19 (s, 6H), 0.98 (s, 9H), 1.30 (t, J = 7.1 Hz, 3H), 2.18 (dd, J = 14.0, 10.2 Hz, 1 H), 2.47 (dd, J= 14.0, 4.8 Hz, 1 H), 2.52 (m, 1 H), 3.75 (s, 3H), 4.14-4.21 (m, 2H), 4.22 (d, J = 2.3 Hz, 1 H), 4.25 (dd, J = 2.8, 1 .9 Hz, 1 H), 6.63 (d, J = 2.8 Hz, 1 H), 6.68 (d, J = 8.8 Hz, 1 H), 6.74 (d, J = 8.5 Hz, 2H), 6.84 (dd, J = 8.8, 2.8 Hz, 1 H), 6.88 (d, J = 8.5 Hz, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 14.4 (CH ₃ ), 18.3 (C), 25.8 (3xCH ₃ ), 32.0 (CH ₂ ), 40.3 (CH), 53.1 (CH), 55.9 (CH ₃ ), 60.4 (CH), 61.6 (CH ₂ ), 115.4 (C), 115.5 (CH), 116.3 (CH), 117.2 (CH), 120.1 (2xCH), 130.2 (2xCH), 131.3 (C), 136.7 (C) , 151.9 (C), 154.4 (C), 172.1 (C); LRMS (ESI+) m/z (%) 497 (100) [M+ H] ⁺ , 454 (35) ; HRMS (ESI+) m/z calc for C ₂₆ H ₃₇ N ₄ 0 ₄ Si 497.2579, found 497.2581 .

(2S,3S,4fl)-Ethyl-3-allyl-4-azido-6-methoxy-1 ,2,3,4-tetrahydroquinoline-2- carboxylate (20)

From imine 2 (300.0 mg, 1 .45 mmol), recently purchased 4-pentenal (187.0 mg, 2.17 mmol) and TMSN ₃ (217.0 mg, 1 .88 mmol), and using method A, the product was obtained after flash chromatography (silica gel, 5% EtOAc/cyclohexane) to give 20 (266.0 mg, 58%); R _f = 0.29 (5% EtOAc/cyclohexane); [a] _D ²⁰ = +188.6 (c = 1.00, CHCI ₃ ); IR v _max (thin film, CH ₂ CI ₂ ) 3405, 2936, 2091 , 1729, 1503, 1226, 1153, 1035 cnr ¹ ; UV: 226, 245; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 1.31 (t, J= 7.1 Hz, 3H), 1.76 (m, 1 H), 1.97 (m, 1 H), 2.34 (m, 1 H), 3.74 (s, 3H), 4.20 (m, 1 H), 4.20-4.31 (m, 2H), 4.40 (d, = 2.3 Hz, 1 H), 4.92 (dd, J= 17.0, 1.4 Hz, 1 H), 5.00 (d, f = 10.1 Hz, 1 H), 5.70 (m, 1 H), 6.64 (d, J = 8.8 Hz, 1 H), 6.67 (d, J = 2.8 Hz, 1 H), 6.81 (dd, J = 8.8, 2.8 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 14.3 (CH ₃ ), 31.0 (CH ₂ ), 38.0 (CH), 52.9 (CH), 55.9 (CH ₃ ), 60.8 (CH), 61 .5 (CH ₂ ), 115.3 (C), 115.4 (CH), 116.2 (CH), 117.1 (CH), 117.7 (CH ₂ ), 135.2 (CH), 137.0 (C), 151.9 (C), 172.0 (C); LRMS (ESI+) m/z (%) 317 (12) [M+ H] ⁺ , 274 (100); HRMS (ESI+) m/z calc for CI ₆ H ₂ IN ₄ 0 ₃ 317.1608, found 317.1600.

(2S,3S,4fl)-Ethyl-3-allyl-4-(allyloxy)-6-methoxy-1 ,2,3,4-tetrahydroquinoline-2- carboxylate (21)

From imine 2 (300.0 mg, 1 .45 mmol), recently purchased 2-pentenal (187.0 mg, 2.17 mmol) and 2-propen-1-ol (109.0 mg, 1.88 mmol), and using method A, the product was obtained after flash chromatography (silica gel, 5% EtOAc/cyclohexane) to give 21 (182.8 mg, 38%); R f = 0.21 (5% EtOAc/cyclohexane); [a] _D ²⁰ = +104.7 (c = 1.02, CHC ); IR v _max (thin film, CH ₂ CI ₂ ) 3408, 2936, 1730, 1638, 1505, 1495, 1210, 1153 cm ¹ ; UV: 245; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 1 .32 (t, J = 7.1 Hz, 3H), 1 .65 (m, 1 H), 1 .91 (m, 1 H), 2.56 (m, 1 H), 3.74 (s, 3H), 4.10 (m, 1 H), 4.11 (m, 1 H), 4.15 (d, J = 2.6 Hz, 1 H), 4.19-4.32 (m, 2H), 4.33 (dd, J= 3.1 , 2.1 Hz, 1 H), 4.90 (ddd, J= 17.0, 3.2, 1.6 Hz, 1 H), 4.96 (m, 1 H), 5.19 (ddd, J = 10.4, 3.0, 1.3 Hz, 1 H), 5.30 (ddd, J= 17.0, 1.6, 1.6 Hz, 1 H), 5.72 (dddd, J= 17.0, 10.4, 7.8, 6.4 Hz, 1 H), 5.95 (dddd, J = 17.0, 10.4, 5.6, 5.6 Hz, 1 H), 6.61 (d, J = 8.7 Hz, 1 H), 6.68 (d, J = 2.9 Hz, 1 H), 6.77 (dd, J = 8.7, 2.9 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 14.3 (CH ₃ ), 30.6 (CH ₂ ), 37.4 (CH), 53.0 (CH), 56.0 (CH ₃ ), 61.3 (CH ₂ ), 69.0 (CH ₂ ), 75.4 (CH), 115.8 (CH), 116.3 (CH), 116.8 (CH), 117.0 (CH), 117.1 (CH), 118.0 (C), 135.4 (CH), 135.9 (CH), 137.1 (C), 151.6 (C), 173.0 (C); LRMS (ESI+) m/z (%) 332 (24) [M+ H] ⁺ , 274 (100); HRMS (ESI+) m/z calc. for Ci ₉ H ₂₆ N0 ₄ 332.1856, found 332.1858. (2S,3S,4fl)-Ethyl-3-allyl-6-methoxy-4-(prop-2-yn-1-yloxy)-1 ,2,3,4- tetrahydroquinoline-2-carboxylate (22)

From imine 2 (300.0 mg, 1 .45 mmol), recently purchased 2-pentenal (187.0 mg, 2.17 mmol) and propargyl alcohol (106.0 mg, 1.88 mmol), and using method A, the product was obtained after flash chromatography (silica gel, 5% EtOAc/cyclohexane) to give 22 (191.0 mg, 40%); R _f = 0.21 (5% EtOAc/cyclohexane); [a] _D ²⁰ = +162.2 (c = 1.06, CHCI ₃ ); IR v _max (thin film, CH ₂ CI ₂ ) 3284, 2942, 1729, 1505, 1229, 1207, 1155 cm ¹ ; UV: 248, 324, 322; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 1.32 (t, J= 7.1 Hz, 3H), 1.68 (m, 1 H), 1.91 (m, 1 H), 2.50 (t, J = 2.4 Hz, 1 H), 2.52 (m, 1 H), 3.76 (s, 3H), 4.18 (dd, J = 16.0, 2.4 Hz, 1 H), 4.18-4.30 (m, 4H), 4.42 (d, J = 2.6 Hz, 1 H), 4.91 (ddd, J = 17.0, 3.2, 1.5 Hz, 1 H), 4.98 (m, 1 H), 5.73 (m, 1 H), 6.63 (d, J = 8.1 Hz, 1 H), 6.77-6.81 (m, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 14.4 (CH ₃ ), 30.7 (CH ₂ ), 37.3 (CH), 52.9 (CH), 54.9 (CH ₂ ), 56.1 (CH3), 61.4 (CH ₂ ), 74.5 (CH), 74.8 (C), 80.4 (CH), 116.0 (CH), 116.7 (C), 116.8 (C), 117.2 (CH), 135.7 (CH), 137.2 (C) , 151.7 (C), 173.0 (C); LRMS (ESI+) m/z(%) 329 (45), 274 (100); HRMS (ESI+) m/z calc for CI ₉ H ₂₄ N0 ₄ 330.1700, found 330.1704.

(2S,3S,4fi)-Ethyl-4-((3-mercaptopropyl)thio)-6-methoxy-3- pentyl-1 ,2,3,4- tetrahydroquinoline-2-carboxylate (23)

Chemical Formula: C _2i H ₃₃ N0 ₃ S ₂ Molecular Weight: 411.6216

From imine 2 (300.0 mg, 1.45 mmol), freshly distilled heptanal (248.0 mg, 2.17 mmol) and 1 ,3 propanedithiol (235.0 mg, 2.17 mmol), and using method A, the product was obtained after flash chromatography (silica gel, 5% EtOAc/cyclohexane) to give 23 (208.5 mg, 35%); R _f = 0.32 (10% EtOAc/cyclohexane); [a] _D ²⁰ = + 51.9 (c = 1.03, CHCI ₃ ); IR v _max (thin film, CH ₂ CI ₂ ) 3398, 2955, 2923, 2850, 1732, 1503, 1467, 1213, 1150, 1039 cm ¹ ; UV: 205, 245, 325; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.85 (t, J = 6.9 Hz, 3H), 1.10 (m, 1 H), 1.15-1.39 (m, 7H), 1.33 (t, J = 7.1 Hz, 3H), 1.96 (m, 2H), 2.34 (m, 1 H), 2.64-2.85 (m, 4H), 3.74 (s, 3H), 3.89 (d, J = 1 .7 Hz, 1 H), 4.21-4.36 (m, 2H), 4.57 (d, J = 2.8 Hz, 1 H), 6.55 (d, J = 8.0 Hz, 1 H), 6.68 (dd, J = 8.0, 2.8 Hz, 1 H), 6.69 (d, J= 2.8 Hz, 1 H); ¹³ C NMR (100 MHz, CDCIs) <5 ppm 14.1 (CH ₃ ), 14.5 (CH ₃ ), 22.6 (CH ₂ ), 23.6 (CH ₂ ), 27.7 (CH ₂ ), 30.1 (CH ₂ ), 31.9 (CH ₂ ), 33.6 (CH ₂ ), 39.0 (CH), 46.2 (CH), 53.7 (CH), 55.9 (CH ₃ ), 61.5 (CH ₂ ), 115.5 (CH), 116.0 (CH), 116.1 (CH), 118.9 (C), 136.7 (C), 152.0 (C), 173.2 (C); LRMS (ESI+) m/z (%) 412 (5) [/W+H] ⁺ , 304 (100); HRMS (ESI+) m/z calc for C _2i H ₃₄ N0 ₃ S ₂ 412.1974, found 412.1966.

Methyl-(2S,3/?,4S)-4-azido-2-ethyl-3-methyl-1,2,3,4-tetra hydroquinoline-6- carboxylate (26)

To a solution of methyl-4-aminobenzoate (1.02 g, 6.75 mmol) and propionaldehyde (1.95 g, 33.1 mmol) at -20°C, D-proline (228.5 mg, 1.985 mmol) was added at -20°C. The reaction mixture was stirred for 20 min at -20°C and kept at the same temperature for two days. Dioxane was added (50 ml.) and TMSN ₃ (1 .3 ml_, 9.92 mmol) and then BF _3. Et ₂ 0 (0.6 ml_, 4.72 mmol) were added at rt and the reaction mixture was stirred at this temperature for 25 min. The reaction was quenched by NaOH (1 N), extracted with EtOAc, dried by MgSC> ₄ , filtered off and concentrated in vacuo. After flash chromatography (silica gel, 10% EtOAc/ cyclohexane), the product 26 was isolated (1.52 g, 82%) as a yellow solid. The product is recrystallized in CH ₂ CI ₂ /heptane to obtain 814 mg (44%) with an enantiomeric excess > 99%; R _f = 0.27 (10% EtOAc in cyclohexane); [a] _D ²⁰ = - 128.3 (c = 0.93, CHCI ₃ ); IR V _max (thin film, CH ₂ CI ): 3427, 3053, 2969, 2093, 1704, 1613, 1436, 1265 cm ¹ ; ¹ H NMR (400 MHz, CDCIs) <5 ppm 0.74 (d, J = 7.1 Hz, 3H), 1.03 (t, J = 7.5 Hz, 3H), 1.53-1.62 (m, 2H), 1 .97 (m, 1 H), 3.42 (m, 1 H), 3.85 (s, 3H), 4.27 (d, J = 2.7 Hz, 1 H), 6.52 (d, J = 8.5 Hz, 1 H), 7.80 (dd, J= 8.5, 2.0 Hz, 1 H), 7.83 (d, J= 2.0 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) d ppm 10.4 (CH ₃ ), 10.5 (CH ₃ ), 25.3 (CH ₂ ), 33.2 (CH), 51.1 (CH ₃ ), 51.8 (CH), 63.8 (CH), 113.5 (CH), 114.5 (C), 118.1 (C), 131.8 (CH), 133.6 (CH), 148.1 (C), 167.2 (C); LMRS (ESI +) m/z (%) 297 (100) [/W+Na] ⁺ ; HRMS (ESI+) m/z calc for Ci ₄ Hi ₈ N ₄ 0 ₂ U [M+ Li] ⁺ 281.1584, found 281.1585. Methyl-(2S,3/?,4S)-1 -acetyl-4-azido-2-ethyl-3-methyl-1 ,2,3,4- tetrahydroquinoline-6-carboxylate (27)

To a stirred solution of tetrahydroquinoline 26 (437.0 mg, 1 .59 mmol) in dry CH2CI2 under argon atmosphere was added dry pyridine (0.39 ml_, 4.78 mmol). Acetyl chloride (0.23 ml_, 3.18 mmol) was added dropwise at 0°C. The mixture was stirred at room temperature overnight. The same procedure was repeated (dry pyridine (0.39 ml_, 4.78 mmol) and acetyl chloride (0.23 ml_, 3.18 mmol) added at 0°C) and the mixture was stirred overnight at room temperature until completion. The solvent was removed and EtOAc was added. The organic layer was washed with brine, dried over MgSC>4 and the solvent was removed in vacuum. The product was obtained after flash chromatography (silica gel, 20% EtOAc/cyclohexane) to give 27 (479.9 mg, 95%); R _f = 0.29 (20% EtOA c/cyclohexane); [O]D ²⁰ = + 307.8 (c = 0.96, CHCI3); IR v _max (thin film, CH2CI2): 1461 , 2967, 2936, 2877, 2095, 1710, 1609, 1497, 1125 cm ¹ ; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.85 (t, J= 7.2 Hz, 3H), 1.07 (m, 1 H), 1.24 (d, J= 6.9 Hz, 3H), 1.61 (m, 1 H), 2.30 (s, 3H), 2.31 (m, 1 H), 3.94 (s, 3H), 4.10 (d, J= 10.3 Hz, 1 H), 4.68 (m, 1 H), 7.35 (d, = 8.5 Hz, 1 H), 7.97 (dd, = 8.5, 2.0 Hz, 1 H), 8.14 (d, J= 2.0 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.7 (CH ₃ ), 16.8 (CH ₃ ), 18.5 (CH ₂ ),

23.8 (CH ₃ ), 39.1 (CH), 52.5 (CH ₃ ), 59.0 (CH), 63.7 (CH), 125.5 (CH), 127.3 (C), 128.2 (C),

129.8 (CH), 130.8 (CH), 141.0 (C), 166.2 (C), 170.1 (C); LRMS (ESI+) m/z (%) 317 (30) [/W+H] ⁺ , 232 (100); HRMS (ESI+) m/z calc. for. C16H21N4O3317.1614, found 317.1608.

Methyl-(2S,3/?,4S)-1 -acetyl-2-ethyl-3-methyl-4-(4-phenyl-1 H-1 ,2,3-triazol-1 -yl)- 1 ,2,3,4-tetrahydroquinoline-6-carboxylate (28) Method B

To a solution of 6-a (300.0 mg, 0.67 mmol) and phenylacetylene (138.0 mg, 1.35 mmol) in CH2CI2 (1 mL) was added Amberlyst Cul.Et3N ^[3] (40.0 mg) at rt. The reaction mixture was stirred at this temperature for 24 h. After completion, the reaction was filtered off, the supported catalyst was washed with CH2CI2 (2 mL), and the solvent was concentrated under vacuum. After flash chromatography (silica gel, 25% EtOAc/ cyclohexane), the product 28 was isolated (318.3 mg, 86%) as a white solid; R _f = 0.12 (30% EtOA c/cyclohexane); [a] _D ²⁰ = +189.9 (c = 1.00, CHCI ₃ ); IR v _max (thin film, CH2CI2): 3478, 3128, 2966, 2877, 2094, 1719, 1664, 1260 cnr ¹ ; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.95 (t, J= 7.2 Hz, 3H), 1 .09 (d, J= 6.7 Hz, 2H), 1 .20-1 .33 (m, 2H), 1 .76 (m, 1 H), 2.41 (s, 3H), 2.65 (m, 1 H), 3.81 (s, 3H), 5.66 (d, J = 11 .2 Hz, 1 H), 7.33 (t, J = 7.4 Hz, 1 H), 7.41 (t, J = 7.4 Hz, 2H), 7.54 (bs, 1 H), 7.64 (d, J = 1 .9 Hz, 1 H), 7.67 (s, 1 H), 7.83 (d, J = 7.4 Hz, 2H), 7.99 (dd, J = 8.6, 1.9 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.8 (CH ₃ ), 16.1 (CH ₃ ), 18.5 (CH ₂ ), 24.1 (CH ₃ ), 40.6 (CH), 52.4 (CH ₃ ), 60.6 (CH), 63.1 (CH), 117.9 (CH), 125.9 (2xCH), 126.9 (CH), 127.6 (2xC), 128.6 (CH), 129.1 (2xCH), 130.1 (CH), 130.3 (CH), 130.4 (C), 140.5 (C), 149.0 (C), 165.9 (C), 170.1 (C); LRMS (ESI+) m/z (%) 441 (100) [/W+Na] ⁺ , 419 (10) [/W+H] ⁺ ; HRMS (ESI+) m/z calc for C ₂₄ H ₂₆ N ₄ 0 ₃ Na 441.1903, found 441.1901.

Methyl-(2S,3S,4fl)-4-azido-2-ethyl-3-methyl-1,2,3,4-tetra hydroquinoline-6- carboxylate (29)

A solution of methyl-4-benzoate (100.0 mg, 0.66 mmol), (S)-(-)-a,a-diphenyl-2- pyrrolidine methanol trimethylsilyl ether (44.0 mg, 0.13 mmol) and 4-nitrophenyl acetic acid (24 mg, 0.13 mmol) in MTBE (1.5 mL) was stirred for 10 min at -10°C. Then propionaldehyde (0.65 mL, 8.61 mmol) was added and stirred for 48 h at -10°C. After the reaction was complete, TMSN ₃ (0.13 mL, 0.99 mmol) and BF _3. Et ₂ 0 (180 pL, 1.39 mmol) were added at -10°C and the mixture was stirred at rt for 4 h. The reaction was quenched with NaOH (2 N) until neutralization, extracted with EtOAc, dried by MgS0 ₄ , filtered off and concentrated by vacuum. After flash chromatography (silica gel, 5% EtOAc/cyclohexane), the product 29 was isolated (106.4 mg, 58.6%); R _f = 0.35 (10% EtOAc/cyclohexane); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.98 (t, J = 7.5 Hz, 3H), 1 .08 (d, J = 6.7 Hz, 3H), 1 .53 (m, 1 H), 1 .72 (m, 1 H), 1 .84 (m, 1 H), 3.24 (ddd, J = 10.3, 7.0, 3.3 Hz, 1 H), 3.85 (s, 3H), 4.39 (d, = 3.1 Hz, 1 H), 6.52 (d, J= 9.1 Hz, 1 H), 7.79 (d, J= 2.0 Hz, 1 H), 7.80 (dd, J= 9.1 , 2.0 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 8.6 (CH ₃ ), 14.3 (CH ₃ ), 25.7 (CH ₂ ), 34.5 (CH), 51.8 (CH ₃ ), 52.9 (CH), 63.7 (CH), 113.5 (CH), 116.7 (C), 117.7 (C), 131.9 (CH), 132.0 (CH), 148.3 (C), 167.2 (C); HRMS (ESI+) m/z calc for Ci ₄ Hi ₈ N ₄ 0 ₂ Li [M+ Li] ⁺ 281.1584, found 281.1585.

Methyl-(2S,3S,4fl)-1-acetyl-4-azido-2-ethyl-3-methyl-1,2, 3,4-tetrahydroquinoline-6- carboxylate (30)

To a stirred solution of tetrahydroquinoline 29 (85.8 mg, 0.31 mmol) in dry CH2CI2 (1 .6 ml.) under argon atmosphere was added dry pyridine (76 mI_, 0.94 mmol). Acetyl chloride (45 mI_, 0.63 mmol) was added dropwise at 0°C. The mixture was stirred at room temperature overnight. The same procedure was repeated (dry pyridine (76 mI_, 0.94 mmol) and acetyl chloride (45 mI_, 0.63 mmol) added at 0°C) and the mixture was stirred for 8 h at room temperature until completion. The solvent was removed and EtOAc was added. The organic layer was washed with brine, dried over MgS0 ₄ and the solvent was removed in vacuum. The product was obtained after flash chromatography (silica gel, 10% EtOAc/cyclohexane) to give 30 (65.2 mg, 65.8%); R _f = 0.20 (10% EtOAc/cyclohexane); ¹ H NMR (400 MHz, CDCh) <5 ppm 0.80 (t, J = 7.4 Hz, 3H), 1 .24 (d, J = 6.7 Hz, 3H), 1 .38 (m, 1 H), 1 .75 (m, 1 H), 1 .83 (m, 1 H), 2.16 (s, 3H), 3.95 (s, 3H), 4.31 (m, 1 H), 4.48 (d, J = 3.3 Hz, 1 H), 7.30 (d, J = 8.3 Hz, 1 H), 7.99 (d, J = 2.0 Hz, 1 H), 8.07 (dd, J = 8.3, 2.0 Hz, 1 H); ¹³ C NMR (100 MHz, CDCh) <5 ppm 9.7 (CH ₃ ), 17.2 (CH ₃ ), 23.2 (CH ₃ ), 27.2 (CH ₂ ), 40.7 (CH), 52.6 (CH ₃ ), 57.4 (CH), 63.8 (CH), 126.7 (CH), 127.8 (C), 129.1 (CH), 131.0 (CH), 132.2 (C), 142.9 (C), 166.2 (C), 170.2 (C); LRMS (ESI+) m/z (%) 339 (25) [/W+Na] ⁺ , 317 (100) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for CI ₆ H ₂ I0 ₃ N ₄ [M+ H] ⁺ 317.1608, found 317.1606.

Methyl-(2S,3S,4fi)-1 -acetyl-2-ethyl-3-methyl-4-(4-phenyl-1 H-1 ,2,3-triazol-1 -yl)-1 ,2,3,4- tetrahydroquinoline-6-carboxylate (31) To a solution of acetyled product 30 (27.5 mg, 0.087 mmol) and phenylacetylene (18 mI_, 0.174 mmol) in dry DCM (0.1 ml.) was added Cul.EtsN (9 mg) according to Method B. The mixture was stirred at RT overnight. Cul.EtsN (6.8 mg) and phenylacetylene were added (18 mI_, 0.174) again, and the reaction was stirring at RT overnight until completion. The reaction was filtered off and washed by DCM and concentrated in vacuo. After flash chromatography (silica gel, 20% EtOAc/cyclohexane), the product was isolated (16.4 mg, 45%).

R _f = 0.23 (20% EtOAc/cyclohexane); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.90 (t, J = 7.2 Hz, 3H), 0.91 (d, J = 7.0 Hz, 3H), 1.53 (m, 1 H), 1.72 (m, 1 H), 2.25 (s, 3H), 2.47 (m, 1 H), 3.81 (s, 3H), 4.39 (m, 1 H), 5.94 (d, J = 6.0 Hz, 1 H), 7.25 (t, J = 7.4 Hz, 1 H), 7.34 (t, J = 7.4 Hz,

2H), 7.47 (m, 1 H), 7.51 (s, 1 H), 7.72 (d, J = 7.4 Hz, 2H), 7.82 (d, J = 2.0 Hz, 1 H), 7.99 (dd, J = 8.5, 2.0 Hz, 1 H); LRMS (ESI+) m/z (%) 441 (35) [/W+Na] ⁺ , 419 (100) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C24H27O3N4 [M+ H] ⁺ 419.2078, found 419.2069. Methyl-(2S,3S,4fi)-4-(4-(2,5,8,11 -tetraoxatetradec-13-yn-1 -yl)-1 H-1 ,2,3-triazol-1 -yl)-1 - acetyl-2-ethyl-3-methyl-1,2,3,4-tetrahydroquinoline-6-carbox ylate (32) Using the same protocol as 31 and starting from 30 (20.5 mg, 0.06 mmol) and dialkyne- PEG (44 mg, 0.19 mmol), and after flash chromatography (silica gel, EtOAc 100%), the compound 32 was obtained (16.2 mg, 46%) :

R _f = 0.34 (2% MeOH/CH ₂ CI ); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.92 (d, J = 6.9 Hz, 3H), 0.93 (t, J = 7.4 Hz, 3H), 1.56 (m, 1 H), 1.79 (m, 1 H), 2.29 (s, 3H), 2.42 (t, J = 2.4 Hz, 1 H), 2.45 (m, 1 H), 3.61-3.72 (m, 12H), 3.88 (s, 3H), 4.19 (d, J= 2.4 Hz, 2H), 4.43 (m, 1 H), 4.67 (s, 2H), 5.91 (d, J = 5.9 Hz, 1 H), 7.30 (s, 1 H), 7.48 (m, 1 H), 7.82 (s, 1 H), 8.04 (dd, J = 8.5, 2.0 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) d ppm 10.2 (CH ₃ ), 16.0 (CH ₃ ), 23.5 (CH ₃ ), 25.9 (CH ₂ ), 38.0 (CH), 52.5 (CH ₃ ), 58.6 (CH ₂ ), 59.8 (CH), 61 .0 (CH), 64.9 (CH ₂ ), 70.0 (CH ₂ ), 70.6 (CH ₂ ), 70.7 (CH ₂ ), 70.7 (CH ₂ ), 70.8 (CH ₂ ), 70.8 (CH ₂ ), 74.7 (CH), 79.9 (C), 122.9 (CH), 126.4 (CH), 127.6 (2xC), 130.4 (CH), 130.7 (CH), 142.1 (C), 145.4 (C), 166.0 (C), 170.7 (C); LRMS (ESI+) m/z (%) 441 (35) [/W+Na] ⁺ , 419 (100) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C ₂₈ H ₃₉ 0 ₇ N ₄ [M+ H] ⁺ 543.2813, found 543.2793.

N-(4'-amino-[1 ,1 '-biphenyl]-4-yl)hex-5-enamide (XA)

Chemical Formula: CI ₃ H ₂₀ N ₂ O Molecular Weight: 280,3710

A solution of 5-hexenoic acid (1 .3 ml_, 7.246 mmol), DMAP (133.0 mg, 1 .09 mmol), and benzidine (2.0 g, 10.87 mmol) in DCM (70 ml.) was cooled at 0°C. DCC (2.24 g, 10.87 mmol) was solubilized in CH ₂ CI ₂ (70 ml.) and this solution was added dropwise to the previous mixture. The reaction mixture was stirred overnight at rt. The mixture was filtered on and washed with HCI (1 N) then with NaHC0 ₃ until pH = 8. Organic layer was washed again with brine, dried over MgS0 ₄ , filtered on and concentrated under vacuum. The product was purified by flash chromatography (30% cyclohexane in 70% mixture of 2% EtOAc in CH ₂ CI ₂ ) to yield the product XA (1.32 g, 65%) as a white solid. R _f = 0.20 (2% EtOAc/CH ₂ CI ₂ ); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 1.84 (m, 2H), 2.15 (m, 2H), 2.36 (t, J = 7.5 Hz, 2H), 4.98-5.08 (m, 2H), 5.80 (m, 1 H), 6.73 (d, J = 8.5 Hz, 2H), 7.37 (d, J = 8.5 Hz, 2H), 7.47 (d, J = 8.7 Hz, 2H), 7.53 (d, J = 8.7 Hz, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 24.8 (CH ₂ ), 33.3 (CH ₂ ), 37.0 (CH ₂ ), 115.6 (2xCH), 115.7 (CH ₂ ), 120.4 (2xCH), 126.9 (2xCH), 127.9 (2xCH), 131.1 (C), 136.5 (C), 137.4 (C), 138.0 (CH), 145.9 (C), 171.3 (C); LRMS (ESI+) m/z (%) 281 (100) [M+ H] ⁺ ; HRMS (ESI+): m/z calc for CI ₈ H ₂ ION ₂ 281 .1648, found 281.1646.

N-(4-((2S,3/?,4S)-4-azido-2-ethyl-3-methyl-1,2,3,4-tetrah ydroquinolin-6- yl)phenyl)hex-5-enamide (XB)

Chemical Formula: C ₂₄ H ₂₉ N ₅ 0

Molecular Weight: 403.52

A solution of N-(4'-amino-[1 ,1'-biphenyl]-4-yl)hex-5-enamide XA (213.0 mg, 0.761 mmol) and D-proline (26.0 mg, 0.226 mmol) in CH ₃ CN (20 ml_) was cooled at -40°C. Propionaldehyde (600.0 mg, 10.33 mmol) was added at -40°C. The reaction mixture was stirred at -40°C for 2 h then at 0°C overnight. TMSN ₃ (114.0 mg, 0.989 mmol) and then BF _3. Et ₂ 0 (0.40 ml_, 3.241 mmol) were added at rt. The reaction mixture is stirred at rt for 1 h30. The reaction is quenched with NaOFI (1 N), concentrated and dissolved in EtOAc. The layer was dried over MgSC> ₄ , filtered on and concentrated under vacuum. The product was purified by flash chromatography (silica gel, 10% to 20% EtOAc/cyclohexane) to obtain XB (194.1 mg, 63%); R _f = 0.37 (20% EtOAc/cyclohexane); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.79 (d, J = 7.1 Hz, 3H), 1 .02 (t, J = 7.4 Hz, 3H), 1 .55 (m, 2H), 1 .84 (m, 2H), 1 .95 (m, 1 H), 2.14 (m, 2H), 2.36 (t, J= 7.5 Hz, 2H), 3.35 (td, J= 7.1 , 2.6 Hz, 1 H), 4.28 (d, J= 2.6 Hz, 1 H), 4.96-5.08 (m, 2H), 5.80 (ddt, J = 16.9, 10.2, 6.7 Hz, 1 H), 6.60 (d, J = 8.4 Hz, 1 H), 7.29 (d, J = 2.0 Hz, 1 H), 7.34 (dd, J = 8.4, 2.0 Hz, 1 H), 7.46 (d, J = 8.5 Hz, 2H), 7.53 (d, J = 8.5 Hz, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.5 (CH ₃ ), 10.6 (CH ₃ ), 24.8 (CH ₂ ), 25.5 (CH ₂ ), 33.3 (CH ₂ ), 33.9 (CH), 37.0 (CH ₂ ), 51.1 (CH), 64.4 (CH), 114.9 (CH), 115.6 (CH ₂ ), 115.9 (C), 120.4 (2xCH), 126.8 (2xCH), 128.3 (CH), 129.3 (C), 129.4 (CH), 136.4 (C), 137.1 (C), 138.0 (CH), 143.6 (C), 171.4 (C); LRMS (ESI+) m/z (%) 361 (100) 404 (10) [M+ H] ⁺ ; HRMS (ESI+): m/z calc for C ₂₄ H ₃ oN ₅ 0404.2450, found 404.2439. N-(4-((2S,3/?,4S)-1-acetyl-4-azido-2-ethyl-3-methyl-1,2,3,4- tetrahydroquinolin- 6-yl)phel)hex-5-enamide (XC)

To a solution of XB (191.5 mg, 0.47 mmol) and pyridine (0.11 mL, 1.43 mmol) in CH ₂ CI ₂ (6 mL) was added acetyl chloride (36 pL, 0.50 mmol) at 0°C. The reaction mixture was stirred at 0°C to rt for 2h30. The reaction was concentrated under vacuum. After flash chromatography (silica gel, 30% EtOAc/cyclohexane), the product XC was isolated (117.8 mg, 84%) as a yellow solid; R _f = 0.20 (30% EtOAc/cyclohexane); [O]D ²⁰ = + 247.5 (c = 1 .03, CHCI ₃ ); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.86 (t, J= 7.2 Hz, 3H), 1.12 (m, 1 H), 1.22 (d, J = 6.9 Hz, 3H), 1 .60 (m, 1 H), 1 .86 (m, 2H), 2.16 (m, 2H), 2.31 (s, 3H), 2.33 (m, 1 H), 2.40 (t, J = 7.5 Hz, 2H), 4.11 (d, J = 10.0 Hz, 1 H), 4.74 (m, 1 H), 5.01 (m, 1 H), 5.06 (m, 1 H), 5.81 (ddt, J = 16.9, 10.0, 6.6 Hz, 1 H), 7.29 (m, 1 H), 7.49 (dd, J = 8.4, 2.1 Hz, 1 H), 7.54 (d, J = 8.6 Hz, 2H), 7.61 (d, J = 2.1 Hz, 1 H), 7.64 (d, J = 8.6 Hz, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.8 (CH ₃ ), 16.9 (CH ₃ ), 18.3 (CH ₂ ), 23.7 (CH ₃ ), 24.7 (CH ₂ ), 33.3 (CH ₂ ), 37.0 (CH ₂ ), 39.5 (CH), 60.6 (CH), 64.1 (CH), 115.6 (CH ₂ ), 120.4 (2xCH), 126.1 (CH), 127.0 (CH), 127.3 (CH), 127.5 (2xCH), 127.6 (C), 127.7 (C), 128.7 (C), 135.5 (C), 138.0 (CH), 138.2 (C), 170.3 (C), 171.6 (C); LMRS (ESI+) m/z (%) 359 (100) 446 (10) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C ₂₆ H _3i N ₅ C> ₂ Na 468.2375, found 468.2383.

N-(4-((2S,3fl,4S)-1 -acetyl-2-ethyl-3-methyl-4-(4-phenyl-1 H-1 ,2,3-triazoM -yl)- 1 ,2,3,4-tetrahydroquinolin-6-yl)phenyl)hex-5-enamide (XD)

According to method B: To a solution of XC (300.0 mg, 0.67 mmol) and phenylacetylene (138.0 mg, 1.35 mmol) in CH ₂ CI ₂ (1 mL) was added Amberlyst Cul.EtsN (40.0 mg) at rt. The reaction mixture was stirred at this temperature for 24 h. After completed, the reaction was filtered off, washed with CH ₂ CI ₂ (2 ml.) and concentrated under vacuum. After flash chromatography (silica gel, 50% EtOAc/cyclohexane), the product XD was isolated (318.3 mg, 86%) as a white solid; R _f = 0.24 (50% EtOAc/cyclohexane); [O]D ²⁰ = + 294.1 (c = 0.97, CHCI ₃ ); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.96 (t, J= 7.3 Hz, 3H), 1 .09 (d, J = 6.8 Hz, 3H), 1.31 (m, 1 H), 1.77 (m, 1 H), 1.82 (m, 2H), 2.10 (m, 2H), 2.35 (m, 2H), 2.41 (s, 3H), 2.64 (m, 1 H), 4.96 (m, 1 H), 5.00 (m, 2H), 5.69 (d, J = 11.3 Hz, 1 H), 5.76 (m, 1 H), 7.12 (s, 1 H), 7.27-7.59 (m, 9H), 7.76 (m, 1 H), 7.78 (d, J = 7.8 Hz, 2H); ¹³ C NMR (100 MHZ, CDCI ₃ ) <5 ppm 10.8 (CH ₃ ), 16.1 (CH ₃ ), 18.3 (CH ₂ ), 24.0 (CH ₃ ), 24.7 (CH ₂ ), 33.2 (CH ₂ ), 37.0 (CH ₂ ), 40.9 (CH), 60.6 (CH), 63.5 (CH), 115.6 (CH ₂ ), 117.8 (CH), 120.3 (2xCH), 125.8 (2xCH), 127.4 (2xCH), 127.5 (C), 127.9 (CH), 128.6 (CH), 128.7 (CH), 128.9 (C), 129.0 (2xCH), 130.1 (C), 130.4 (CH), 134.9 (C), 138.0 (CH), 138.1 (C), 138.4 (C), 149.0 (C), 170.2 (C), 171.5 (C); LRMS (ESI+) m/z (%) 570 (100) 548 (10) [M+H] ⁺ ; HRMS (ESI+) m/z calc for C ₃₄ H ₃₇ N ₅ 0 ₂ Na 570.2845, found 570.2842.

N-(4-((2S,3fl,4S)-1 -acetyl-2-ethyl-3-methyl-4-(4-pentyl-1 H-1 ,2,3-triazoM -yl)- 1 ,2,3,4-tetrahydroquinolin-6-yl)phenyl)hex-5-enamide (XE)

From XC (100.0 mg, 0.22 mmol), 1 -heptyne (43.0 mg, 0.45 mmol), Amberlyst Cul.Et ₃ N (40.0 mg), CH ₂ CI ₂ (1 ml_), and using the method B, the product was obtained after flash chromatography (silica gel 30% to 50% EtOAc/cyclohexane) to give XE (87.0 mg, 71.5%); R _f = 0.25 (50% EtOAc/cyclohexane); [a] _D ²⁰ = + 234.6 (c = 0.68, CHCI ₃ ); ¹ H NMR (400 MHz, CDCI ₃ ) d ppm 0.84 (t, J = 6.8 Hz, 3H), 0.95 (t, J = 7.6 Hz, 3H), 1 .05 (d, J = 6.8 Hz, 3H), 1 .25-1.35 (m, 5H), 1.64 (m, 2H), 1.73 (m, 1 H), 1.84 (m, 2H), 2.14 (m, 2H), 2.38 (m, 2H), 2.39 (s, 3H), 2.56 (m, 1 H), 2.68 (m, 2H), 4.95-5.07 (m, 3H), 5.61 (d, J = 11 .3 Hz, 1 H), 5.79 (m, 1 H), 7.05 (s, 1 H), 7.15 (s, 1 H), 7.17 (m, 1 H), 7.35 (d, J = 8.5 Hz, 2H), 7.50 (dd, J = 8.4, 2.0 Hz, 1 H), 7.56 (d, J = 8.5 Hz, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.8 (CH ₃ ), 14.1 (CH ₃ ), 16.1 (CH ₃ ), 18.2 (CH ₂ ), 22.5 (CH ₂ ), 23.9 (CH ₃ ), 24.7 (CH ₂ ), 26.0 (CH ₂ ), 29.1 (CH ₂ ), 31.6 (CH ₂ ), 33.3 (CH ₂ ), 37.0 (CH ₂ ), 40.8 (CH), 58.2 (CH), 63.2 (CH), 115.6 (CH ₂ ), 118.7 (CH), 120.3 (2xCH), 126.3 (CH), 127.3 (CH), 127.4 (3xCH), 127.7 (C), 135.1 (C), 138.0 (CH, C), 138.1 (C), 138.3 (C), 149.9 (C), 170.2 (C), 171.5 (C); LRMS (ESI+) m/z (%) 564 (100) 542 (40) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C ₃₃ H ₄₄ N ₅ O ₂ 542.3495, found 542.3489.

Methyl (2S,3fl,4S)-1 -acetyl-2-ethyl-3-methyl-4-(4-pentyl-1 H-1 ,2,3-triazol-1 -yl)- 1 ,2,3,4-tetrahydroquinoline-6-carboxylate (XF)

From 27 (100.0 mg, 0.32 mmol), 1-heptyne (61.0 mg, 0.63 mmol), Amberlyst Cul.Et ₃ N (40.0 mg), CH2CI2 (1 ml_), and using the method B, the product was obtained after flash chromatography (silica gel, 30% EtOAc/ cyclohexane) to give XF (98.0 mg, 75%); R _f = 0.10 (30% EtOAc/cyclohexane) ; [a] _D ²⁰ = +219.7 (c = 0.76, CHCI ₃ ); ¹ H NMR (400 MHz, CDCI3) <5 ppm 0.88 (t, J= 7.4 Hz, 3H), 0.93 (t, J= 7.2 Hz, 3H), 1 .04 (d, J= 6.8 Hz, 3H), 1 .24 (m, 1 H), 1 .28-1 .36 (m, 4H), 1 .66 (m, 2H), 1 .73 (m, 1 H), 2.38 (s, 3H), 2.57 (m, 1 H), 2.71 (t, J = 7.6 Hz, 2H), 3.83 (s, 3H), 4.69 (bs, 1 H), 5.57 (d, J = 11 .3 Hz, 1 H), 7.14 (s, 1 H), 7.49 (s, 1 H), 7.57 (d, J = 2.1 Hz, 1 H), 7.97 (d, J = 8.5, 2.1 Hz, 1 H); ¹³ C NMR (125 MHz, CDCI3) <5 ppm 1 0.8 (CH ₃ ), 14.1 (CH ₃ ), 16.1 (CH ₃ ), 18.4 (CH ₂ ), 22.5 (CH ₂ ), 24.0 (CH ₃ ), 26.0 (CH ₂ ), 29.0 (CH ₂ ), 31.6 (CH ₂ ), 40.4 (CH), 52.4 (CH ₃ ), 59.5 (CH), 62.8 (CH), 118.9 (CH), 125.8 (CH), 127.2 (C), 127.6 (C), 130.0 (CH), 130.1 (CH), 140.5 (C), 149.8 (C), 165.9 (C), 170.1 (C); LRMS (ESI+) m/z (%) 274 (100) 413 (55) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C23H33N4O3413.2553, found 413.2551 .

N-(4-((2S,3S,4S)-1-acetyl-4-amino-2-ethyl-3-methyl-1,2,3, 4-tetrahydroquinolin- 6-yl)phenyl)hex-5-enamide (XG)

To a solution of XC (300.0 mg, 0.67 mmol) in methanol (6 ml.) were added EtsN (0.47 ml_, 3.37 mmol) and 1 ,3-propanedithiol (0.34 ml_, 3.37 mmol) at rt. The reaction mixture was added at this temperature for 24 h. After completion, the reaction was diluted in methanol and H2O, acidified by HCI 1 M, washed with ether. The organic layer was basified with NaOH 1 M, extracted by EtOAc four times, dried by MgSC>4, filtered off and concentrated under vacuum. The product XG was obtained (240.9 mg, 85%) as a yellow solid; R _f = 0.24 (4% MeOH/CH ₂ CI ; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.75-0.85 (t, J = 7.3 Hz, 3H), 1.09 (m, 1 H), 1.18 (d, J = 6.9 Hz, 3H), 1.55 (m, 1 H), 1.83 (m, 2H), 2.14 (m, 2H), 2.29 (s, 3H), 2.39 (m, 2H), 2.86 (m, 1 H), 3.68 (d, J = 10.0 Hz, 1 H), 4.78 (m, 1 H), 5.00 (m, 1 H), 5.04 (m, 1 H), 5.81 (m, 1 H), 7.21 (m, 1 H), 7.40 (d, J = 2.0 Hz, 1 H) 7.48-7.64 (m, 4H), 7.73 (m, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) d ppm 10.7 (CH ₃ ), 16.8 (CH ₃ ), 18.2 (CH ₂ ), 23.7 (CH ₃ ), 24.8 (CH ₂ ), 33.3 (CH ₂ ), 36.9 (CH ₂ ), 42.1 (CH), 53.9 (CH), 58.4 (CH), 115.5 (CH ₂ ), 120.4 (2xCH), 125.7 (CH), 126.9 (CH), 127.3 (3xCH, C), 135.6 (C), 137.8 (C), 137.9 (2xC), 138.0 (CH), 170.3 (C), 171.9 (C); LRMS (ESI-) m/z (%) 454 (100) 418 (40) [M- H] ; HRMS (ESI-) m/z calc for C ₂₆ H ₃₂ N ₃ 0 ₂ 418.2495, found 418.2476.

Methyl-(2S,3S,4S)-1-acetyl-4-amino-2-ethyl-3-methyl-1,2,3 ,4- tetrahydroquinoline-6-carboxylate (XH)

To a solution of 27 (843.2 mg, 2.66 mmol) in ethanol (40 ml.) was added Pd/C (290.0 mg, 2.66 mmol). The reaction mixture was stirred at rt under H ₂ for the night. The reaction was filtered off under celite, washed with ethanol and concentrated under vacuum. The product XH was isolated (744.5 mg, 96.2%); R _f = 0.33 (5% MeOH/CH ₂ CI ₂ ); ¹ H NMR (400 MHz, CDsOD) <5 ppm 0.82 (t, J = 7.2 Hz, 3H), 1 .01 (m, 1 H), 1 .30 (d, J = 6.9 Hz, 3H), 1 .68 (m, 1 H), 2.31 (m, 1 H), 2.34, (s, 3H), 3.93 (s, 3H), 4.31 (d, J = 9.5 Hz, 1 H), 4.66 (m, 1 H), 7.64 (d, J = 8.3 Hz, 1 H), 8.04 (d, J = 8.3, 1.6 Hz, 1 H), 8.27 (d, J = 1 .6 Hz, 1 H); ¹³ C NMR <5 ppm (100 MHz, CD ₃ OD) 10.5 (CH ₃ ), 16.4 (CH ₃ ), 19.2 (CH ₂ ), 23.5 (CH ₃ ), 39.5 (CH), 52.9 (CH ₃ ), 54.2 (2xCH), 127.8 (CH), 128.9 (C), 131.5 (2xCH), 132.7 (C), 142.3 (C), 167.2 (C), 172.4 (C); LRMS (ESI+) m/z (%) 232 (100); HRMS (ESI+) m/z calc for Ci ₆ H ₂₂ N ₂ 0 ₃ Na 313.1528, found 313.1526 lsopropyl-((2S,3S,4S)-1-acetyl-2-ethyl-6-(4-(hex-5-enamido)p henyl)-3-methyl- 1 ,2,3,4-tetrahydroquinolin-4-yl)carbamate (XI)

To a solution of XG (220.0 mg, 0.52 mmol) and DIPEA (0.27 ml_, 1.57 mmol) in CH ₂ CI ₂ (6 ml.) under N ₂ at rt was added isopropyl chloroformate 2 M in toluene (0.39 ml_, 0.79 mmol). The reaction mixture was stirred at this temperature for 2 h. The reaction was concentrated under vacuum. EtOAc (11 ml.) and H ₂ 0 (4 ml.) were added and the organic layer was washed with H ₂ 0 (3 ml_), with saturated aqueous solution of NaCI (1 .5 ml_), dried over MgSC> ₄ , filtered off and concentrated under vacuum. After flash chromatography (silica gel, 20% to 50% EtOAc/cyclohexane), the product XI (188.9 mg, 71%) was isolated as a white solid. R _f = 0.35 (50% EtOAc/cyclohexane); [a] _D ²⁰ = + 111 .3 (c = 1 .02, CHCI ₃ ); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.88 (t, J = 7.2 Hz, 3H), 1 .15 (d, J = 6.8 Hz, 3H), 1 .27 (d, J = 6.4 Hz, 6H), 1.28 (m, 1 H), 1.65 (m, 1 H), 1.86 (m, 2H), 2.06 (m, 1 H), 2.17 (m, 2H), 2.30 (s, 3H), 2.39 (t, J= 7.5 Hz, 2H), 4.60 (m, 1 H), 4.72 (m, 1 H), 4.94- 5.11 (m, 3H), 5.82 (ddt, J = 16.9, 10.2, 6.7 Hz, 1 H), 7.21 (m, 1 H), 7.43 (dd, J= 8.3, 2.1 Hz, 1 H), 7.51 (d, J= 8.5 Hz, 2H), 7.56 (m, 1 H), 7.58 (d, J = 8.5 Hz, 2H); ¹³ C NMR (100 MHz, CD ₃ OD) d ppm 11.1 (CH ₃ ), 16.6 (CH ₃ ), 19.0 (CH ₂ ), 22.6 (2xCH ₃ ), 23.7 (CH ₃ ), 26.2 (CH ₂ ), 34.5 (CH ₂ ), 37.5 (CH ₂ ), 40.6 (CH), 54.2 (2xCH), 69.7 (CH), 115.9 (CH ₂ ), 121.6 (2xCH), 126.7 (CH), 127.3 (C), 127.5 (CH), 128.1 (3xCH), 137.0 (C), 139.2 (CH, C), 139.6 (2xC), 159.8 (C), 172.9 (C), 174.5 (C); LRMS (ESI+) m/z (%) 528 (100) 506 (10) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C ₃ oH ₃₉ N ₃ 0 ₄ Na 528.2838, found 528.2839.

Methyl-(2S,3S,4S)-1-acetyl-2-ethyl-4-((isopropoxycarbonyl )amino)-3-methyl- 1 ,2,3,4-tetrahydroquinoline-6-carboxylate (XJ)

Chemical formula: C2 ₀ H2 ₈ N2O ₅ Molecular weight: 376.45 To a solution of XH (70.0 mg, 0.24 mmol) and DIPEA (0.12 mL, 0.70 mmol) in CH2CI2 (2 mL) under N ₂ at rt was added isopropyl chloroformate 2 M in toluene (0.18 mL, 0.36 mmol). The reaction mixture was stirred at this temperature for 24 h. The reaction was concentrated under vacuum. EtOAc (8 mL) and H ₂ 0 (4 mL) were added and the organic layer was washed with H ₂ 0 (3 mL), with saturated aqueous solution of NaCI (1 .5 mL), dried over MgSC>4, filtered off and concentrated under vacuum. The product was obtained after flash chromatography (silica gel, 20% to 30% EtOAc/cyclohexane) to give XJ (70.9 mg, 78%); FL = 0.37 (50% EtOAc/cyclohexane); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.87 (t, J = 7.3 Hz, 3H), 1.13 (m, 1 H), 1.14 (d, J = 6.9 Hz, 3H), 1.27 (d, J = 6.2 Hz, 3H), 1.31 (d, J= 6.2 Hz, 3H), 1.65 (m, 1 H), 2.07 (m, 1 H), 2.28 (s, 3H), 3.89 (s, 3H), 4.62 (m, 1 H), 4.81 (t, J= 9.9 Hz, 1 H), 5.0 (m, 1 H), 7.34 (m, 1 H), 7.89 (dd, J = 8.5, 2.0 Hz, 1 H), 8.05 (d, J = 2.0 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI3) d ppm 10.9 (CH ₃ ), 16.4 (CH ₃ ), 18.4 (CH ₂ ), 22.3 (2xCH ₃ ), 24.2 (CH ₃ ), 40.5 (CH), 52.4 (CH ₃ ), 52.8 (2xCH), 69.0 (CH), 125.4 (C), 127.2 (CH), 129.0 (CH), 130.6 (CH, C), 140.5 (C), 157.2 (C), 166.5 (C), 170.2 (C); LRMS (ESI+) m/z (%) 399 (100) 377 (10) [/W+H] ⁺ ; HRMS (ESI+) m/z calc for C20H29N2O5377.2076, found 377.2069.

N-(4-((2S,3S,4S)-1-acetyl-4-((5-chloropynmidin-2-yl)amino )-2-ethyl-3-methyl- 1 ,2,3,4-tetrahydroquinolin-6-yl)phenyl)hex-5-enamide (XK)

To a solution of XG (30.5 mg, 0.073 mmol) and 2,5-dichloropyrimidine (22.0 mg, 0.145 mmol) in DMSO (0.4 mL) was added DIPEA (28.0 mg, 0.218 mmol). The reaction mixture was stirred under microware at 160°C for 1 h30. After cooling to rt EtOAc and H2O were added. The reaction was extracted by EtOAc, the organic layer was dried by MgS04, filtered off and concentrated under vacuum. After flash chromatography (silica gel, 20% to 50% EtOAc/cyclohexane), the product XK was obtained (15.0 mg, 38.7%); R _f = 0.36 (50% EtOAc/cyclohexane); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.91 (t, J= 7.2 Hz, 3H), 1.11 (d, J = 6.9 Hz, 3H), 1 .29 (m, 1 H), 1 .70 (m, 1 H), 1 .85 (m, 2H), 2.16 (m, 2H), 2.25 (m, 1 H), 2.33 (s, 3H), 2.37 (t, J = 7.5 Hz, 2H), 4.86 (m, 1 H), 5.01 (m, 1 H), 5.05 (m, 1 H), 5.15 (m, 1 H), 5.81 (ddt, J = 17.0, 10.0, 6.7 Hz, 1 H), 7.29 (s, 1 H), 7.39-7.44 (m, 3H), 7.48 (m, 1 H), 7.54 (d, J = 8.2 Hz, 2H), 8.18 (s, 2H); ¹³ C NMR (100 MHz, CDCI3) <5 ppm 10.9 (CH ₃ ), 16.5 (CH ₂ ), 18.4 (CH ₂ ), 23.9 (CH ₃ ), 24.8 (CH ₂ ), 33.3 (CH ₂ ), 37.1 (CH ₂ ), 40.6 (CH), 53.4 (CH), 60.6 (CH), 115.8 (CH ₂ ), 119.5 (C), 120.3 (2xCH), 126.1 (CH), 126.7 (C), 127.6 (4xCH), 135.1 (C), 137.5 (C), 137.8 (C) 138.0 (CH, C), 156.5 (C), 161.3 (2xCH), 170.3 (C), 171.3 (C); LRMS (ESI+) m/z (%) 532 (100) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C ₃ oH ₃₅ N ₅ 0 ₂ CI 532.2479, found 532.2485.

Methyl (2S,3S,4S)-1-acetyl-4-((5-chloropynmidin-2-yl)amino)-2-ethyl -3-methyl- 1 ,2,3,4-tetrahydroquinoline-6-carboxylate (XL)

To a solution of XH (100.0 mg, 0.34 mmol) and 2,5-dichloropyrimidine (103.0 mg, 0.69 mmol) in DMSO (1 mL) was added DIPEA (0.17, 1 .0 mmol). The reaction mixture was stirred under microware at 160°C for 1 h30. After cooling to rt EtOAc and H ₂ 0 were added. The reaction was extracted by EtOAc, the organic layer was dried by MgS0 ₄ , filtered off and concentrated under vacuum. The product was obtained after flash chromatography (silica gel, 30% to 50% EtOAc/cyclohexane) to give XL (13.5 mg, 10%); R _f = 0.36 (50% EtOAc/cyclohexane); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.89 (t, J = 7.3 Hz, 3H), 1.10 (d, J = 6.9 Hz, 3H), 1 .21 (m, 1 H), 1 .69 (m, 1 H), 2.27 (m, 1 H), 2.31 (s, 3H), 3.85 (s, 3H), 4.64 (m, 1 H), 5.51 (d, J = 9.7 Hz, 1 H), 7.34 (s, 1 H), 7.89 (dd, J = 8.5, 2.1 Hz, 1 H), 7.99 (d, J = 2.1 Hz, 1 H), 8.20 (s, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.8 (CH ₃ ), 16.4 (CH ₃ ), 18.5 (CH ₂ ), 23.9 (CH ₃ ), 39.9 (CH), 52.3 (CH ₃ ), 53.1 (CH), 60.5 (CH), 119.7 (CH, C), 125.4 (C), 127.1 (C), 128.8 (CH), 130.2 (CH), 140.8 (C), 156.5 (2xCH), 161.1 (C), 166.5 (C), 170.2 (C); LRMS (ESI+) m/z (%) 232 (100) 403 (45) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C ₂₀ H ₂₄ N ₄ O ₃ CI 403.1537, found 403.1534. Methyl (2S,3/?,4S)-1 -acetyl-2-ethyl-4-(4-(4-methoxyphenyl)-1 H-1 ,2,3-triazol-1 - yl)-3-methyl-1 ,2,3,4-tetrahydroquinoline-6-carboxylate (XM) From 27 (106.0 mg, 0.33 mmol), 4-ethynylanisole (89.0 mg, 0.67 mmol), Amberlyst

Cul.EtsN (1 0.0 mg), CH2CI2 (0.4 ml_), and using the method B, the product was obtained after flash chromatography (silica gel, 40% EtOAc/cyclohexane) to give XM (118.0 mg, 78.4%); R r = 0.25 (40% EtOAc/cyclohexane); [a] _D ²⁰ = +146.6 (c = 1.05, CHCI ₃ ); ¹ H NMR (400 MHz, CDCI3) <5 ppm 0.95 (t, J = 7.2 Hz, 3H), 1 .08 (d, J = 6.8 Hz, 3H), 1 .25 (m, 1 H), 1 .76 (m, 1 H), 2.40 (s, 3H), 2.66 (m, 1 H), 3.80 (s, 3H), 3.82 (s, 3H), 4.70 (m, 1 H), 5.64 (d, J

= 11 .2 Hz, 1 H), 6.93 (d, J = 8.8 Hz, 2H), 7.55 (m, 1 H), 7.62 (s, 1 H), 7.64 (s, 1 H), 7.74 (d, J = 8.8 Hz, 2H), 7.98 (dd, J= 8.5, 2.0 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.7 (CH ₃ ), 16.0 (CH ₃ ), 18.4 (CH ₂ ), 24.0 (CH ₃ ), 40.4 (CH), 52.3 (CH ₃ ), 55.4 (CH ₃ ), 60.5 (CH), 62.9 (CH), 114.4 (2xCH), 117.1 (CH), 123.1 (CH), 125.8 (CH), 126.9 (C), 127.1 (2xCH), 127.5 (C), 130.0 (CH), 130.1 (CH), 140.5 (C), 148.7 (C), 159.9 (C), 165.8 (C), 170.1 (C); LRMS (ESI+) m/z (%) 449 (100) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C25H29N4O4 449.2189, found 449.2190.

Methyl (2S,3/?,4S)-1-acetyl-2-ethyl-3-methyl-4-(4-(4-(tnfluoromethy l)phenyl)- 1 H-1 ,2,3-triazol-1-yl)-1 ,2,3,4-tetrahydroquinoline-6-carboxylate (XN)

From 27 (110.0 mg, 0.35 mmol), 1 -ethynyl-4-(trifluoromethyl)benzene (118.0 mg, 0.70 mmol), Amberlyst Cul.EtsN (10.0 mg), CH2CI2 (0.4 ml_), and using the method B, the product was obtained after flash chromatography (silica gel, 30% EtOAc/cyclohexane) to give XN (123.0 mg, 73%); R _f = 0.15 (30% EtOAc/cyclohexane); [a] _D ²⁰ = +123.3 (c = 1.23, CHCI3); ¹ H NMR (400 MHz, CDCI3) <5 ppm 0.96 (t, J= 7.2 Hz, 3H), 1.11 (d, J= 6.8 Hz, 3H), 1 .26 (m, 1 H), 1 .78 (m, 1 H), 2.41 (s, 3H), 2.67 (m, 1 H), 3.81 (s, 3H), 4.70 (m, 1 H), 5.68 (d, J = 11 .3 Hz, 1 H), 7.51 (m, 1 H), 7.64 (d, J= 2.0 Hz, 1 H), 7.66 (d, J= 8.2 Hz, 2H), 7.78 (s, 1 H), 7.94 (d, J = 8.2 Hz, 2H), 8.00 (dd, J = 8.6, 2.0 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.7 (CH ₃ ), 16.1 (CH ₃ ), 18.5 (CH ₂ ), 24.0 (CH ₃ ), 40.7 (CH), 52.4 (CH ₃ ), 59.6 (CH), 63.3 (CH), 118.8 (CH), 124.2 (CF ₃ , q, J = 272 Hz), 125.9 (2xCH, q, J = 4 Hz), 126.0 (2xCH), 126.7 (2xC), 127.7 (C), 130.0 (CH), 130.2 (CH), 130.3 (C, q, J = 32 Hz), 133.9 (C), 140.6 (C), 147.6 (C), 165.8 (C), 170.1 (C); LRMS (ESI+) m/z (%) 487 (100) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C25H26N4O3F3487.1957, found 487.1959.

Methyl-(2S,3R,4S)-1 -acetyl-4-(4-cyclohexyl-1 H-1 ,2,3-triazol-1 -yl)-2-ethyl-3- methyl-1 ,2,3,4-tetrahydroquinoline-6-carboxylate (XO)

From 27 (67.5 mg, 0.21 mmol), cyclohexylacetylene (46.0 mg, 0.43 mmol), Amberlyst Cul.EtsN (10.0 mg), CH2CI2 (0.4 ml_), and using the method B, the product was obtained after flash chromatography (silica gel, 30% EtOAc/cyclohexane) to give XO (67.0 mg, 74%); R,= 0.16 (30% EtOAc/cyclohexane); [a] _D ²⁰ = +158.6 (c = 1.20, CHCI ₃ ); ¹ H NMR (400 MHz, CDCI3) <5 ppm 0.93 (t, J = 7.2 Hz, 3H), 1.03 (d, J = 6.8 Hz, 3H), 1.16-1.45 (m, 5H), 1 .66-1 .86 (m, 5H), 2.05 (m, 2H), 2.38 (s, 3H), 2.57 (m, 1 H), 2.75 (m, 1 H), 3.8 (s, 3H), 4.68 (s, 1 H), 5.56 (d, J = 11 .3 Hz, 1 H), 7.10 (s, 1 H), 7.48 (m, 1 H), 7.58 (d, J = 2.0 Hz, 1 H), 7.97 (dd, J = 8.6, 2.0 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) d ppm 10.8 (CH ₃ ), 16.1 (CH ₃ ), 18.5 (CH ₂ ), 24.0 (CH ₃ ), 26.2 (CH ₂ ), 26.3 (CH ₂ ), 27.1 (CH ₂ ), 33.0 (CH ₂ ), 33.1 (CH ₂ ), 35.6 (CH), 40.5 (CH), 52.4 (CH ₃ ), 59.9 (CH), 62.8 (CH), 117.5 (CH), 125.8 (CH), 127.3 (C), 127.6 (C), 130.1 (C), 130.2 (C), 140.5 (C), 155.1 (C), 166.0 (C), 170.1 (C); LRMS (ESI+) m/z (%) 425 (100) [/W+H] ⁺ ; HRMS (ESI+) m/z calc for C24H33N4O3425.2533, found 425.2562. Methyl-(2S,3/?,4S)-1-acetyl-2-ethyl-3-methyl-4-(4-((2-(prop- 2-yn-1- yloxy)ethoxy)methyl)-1H-1 ,2,3-triazol-1-yl)-1 ,2,3,4-tetrahydroquinoline-6-carboxylate (XP)

Method C

To a stirred solution of 27 (100.0 mg, 0.32 mmol), 1 ,2-bis(prop-2-yn-1-yloxy)ethane (131.0 mg, 0.95 mmol) in 1 :1 H ₂ 0/f-BuOH (4 ml.) was added CuS0 ₄ .5H ₂ 0 (16.0 mg, 0.06 mmol), and sodium ascorbate (63.0 mg, 0.22 mmol) and the mixture was stirred under argon at rt for 48 h. The product was extracted with EtOAc, dried (MgSC>4) and the solvent was removed in vacuum. The product was obtained after flash chromatography (silica gel, 100% EtOAc) to give XP (70.2 mg, 49%); R _f = 0.44 (80% EtOAc/cyclohexane); ¹ H NMR (400 MHz, CDCh) <5 ppm 0.92 (t, J = 7.2 Hz, 3H), 1 .05 (d, J = 6.8 Hz, 3H), 1 .22 (m, 1 H), 1 .73 (m, 1 H), 2.38 (s, 3H), 2.42 (t, J= 2.4 Hz, 1 H), 2.59 (m, 1 H), 3.68-3.75 (m, 4H), 3.84 (s, 3H), 4.17 (d, J = 2.4 Hz, 2H), 4.71 (m, 3H), 5.59 (d, J = 11 .3 Hz, 1 H), 7.46 (s, 1 H), 7.48 (m, 1 H), 7.58 (d, J = 2.0 Hz, 1 H), 7.97 (dd, J = 8.6, 2.0 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) d ppm 10.7 (CH ₃ ), 16.1 (CH ₃ ), 18.4 (CH ₂ ), 24.0 (CH ₃ ), 40.5 (CH), 52.4 (CH ₃ ), 58.6 (CH ₂ ), 59.3 (CH), 63.1 (CH), 65.1 (CH ₂ ), 69.1 (CH ₂ ), 70.0 (CH ₂ ), 74.8 (CH), 79.7 (C), 120.9 (CH), 125.8 (CH), 127.0 (C), 127.6 (C), 130.1 (CH), 130.2 (CH), 140.6 (C), 146.6 (C), 165.9 (C), 170.1 (C); LRMS (ESI+) m/z (%) 455 (100) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C ₂₄ H ₃ IN ₄ 05455.2289, found 455.2282. Dimethyl 4,4'-((((oxybis(ethane-2,1-diyl))bis(oxy))bis(methylene))bis (1 H-1,2,3- triazole-4,1 -diyl))(2S,2'S, 3R, 3'R, 4S,4'S)-bis(1 -acetyl-2-ethyl-3-methyl-1 ,2,3,4- tetrahydroquinoline-6-carboxylate) (XQ)

From 27 (100.0 mg, 0.32 mmol), 3-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)prop-1 - yne (173.0 mg, 0.95 mmol), 1 :1 H ₂ 0/f-Bu0H (4 ml_), CuS0 ₄ .5H ₂ 0 (16.0 mg, 0.06 mmol), and sodium ascorbate (63.0 mg, 0.22 mmol) and using method C, the product was obtained after flash chromatography (silica gel, 100% EtOAc) to give XQ (69.5 mg, 44%); R _f = 0.44 (4% MeOH/ CH ₂ CI ₂ ); [a] _D ²⁰ = +97.8 (c = 0.92, CHCI ₃ ); IR v _max (thin film, CH ₂ CI ₂ ) 3421 , 2963, 2879, 2372, 2094, 1719, 1664, 1260 cm ¹ ; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.92 (t, J = 7.2 Hz, 3H), 1.04 (d, J = 6.8 Hz, 3H), 1.22 (m, 1 H), 1.73 (m, 1 H), 2.38 (s, 3H), 2.41 (t, J = 2.4 Hz, 1 H), 2.59 (m, 1 H), 3.60-3.77 (m, 8H), 3.84 (s, 3H), 4.17 (d, J = 2.4 Hz, 2H), 4.71 (m, 3H), 5.59 (d, J = 11 .2 Hz, 1 H), 7.44 (m, 1 H), 7.46 (s, 1 H), 7.57 (d, J = 2.0 Hz, 1 H), 7.97 (d, J= 8.5, 2.0 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.7 (CH ₃ ), 16.1 (CH ₃ ), 18.4 (CH ₂ ), 24.0 (CH ₃ ), 40.4 (CH), 52.4 (CH ₃ ), 58.5 (CH ₂ ), 59.4 (CH), 63.1 (CH), 65.1 (CH ₂ ), 69.2 (CH ₂ ), 70.1 (CH ₂ ), 70.5 (CH ₂ ), 70.7 (CH ₂ ), 74.7 (CH), 79.8 (C), 121.0 (CH), 125.8 (CH), 127.0 (C), 127.6 (C), 130.1 (CH), 130.2 (C), 140.6 (C), 146.6 (C), 165.9 (C), 170.1 (C); LRMS (ESI+) m/z (%) 521.2 (100) [/W+Na] ⁺ , 499 (60) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C ₂₆ H ₃₅ N ₄ 0 ₆ 499.2551 , found 499.2548.

Methyl-(2S,3ff,4S)-4-(4-(2,5,8,11-tetraoxatetradec-13-yn- 1-yl)-1 H-1 ,2,3-triazol- 1-yl)-1-acetyl-2-ethyl-3-methyl-1 ,2,3,4-tetrahydroquinoline-6-carboxylate (XR) From 27 (100.0 mg, 0.32 mmol), 4,7,10,13-tetraoxahexadeca-1 ,15-diyne (219.0 mg,

0.95 mmol), 1 :1 H ₂ 0/f-Bu0H (4 mL), CuS0 ₄ .5H ₂ 0 (16.0 mg, 0.06 mmol), and sodium ascorbate (63.0 mg, 0.22 mmol) and using method C, the product was obtained after flash chromatography (silica, 100% EtOAc) to give XR (72.7 mg, 42%); R _f = 0.31 (4% MeOH/CH ₂ CI ₂ ); [a] _D ²⁰ = +195.7 (c = 0.44, CHCI ₃ ); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.92 (t, J = 7.2 Hz, 3H), 1 .04 (d, J = 6.8 Hz, 3H), 1 .22 (m, 1 H), 1 .73 (m, 1 H), 2.38 (s, 3H), 2.42

(t, J= 2.4 Hz, 1 H), 2.60 (m, 1 H), 3.63-3.72 (m, 12H), 3.84 (s, 3H), 4.18 (d, J = 2.4 Hz, 2H), 4.70 (m, 3H), 5.58 (d, J = 11 .3 Hz, 1 H), 7.47 (m, 2H), 7.57 (d, J = 2.0 Hz, 1 H), 7.97 (d, J = 8.6, 2.0 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) d ppm 10.7 (CH ₃ ), 16.1 (CH ₃ ), 18.4 (CH ₂ ), 24.0 (CH ₃ ), 40.4 (CH), 52.4 (CH ₃ ), 58.5 (CH ₂ ), 59.4 (CH), 63.1 (CH), 65.1 (CH ₂ ), 69.2 (CH ₂ ), 70.1 (CH ₂ ), 70.5 (CH ₂ ), 70.7 (2xCH ₂ ), 70.8 (CH ₂ ), 74.7 (CH), 79.8 (C), 121.0 (CH), 125.8

(CH), 127.0 (C), 127.6 (C), 130.1 (CH), 130.2 (C), 140.6 (C), 146.6 (C), 165.9 (C), 170.1 (C); LRMS (ESI+) m/z (%) 566 (100) [/W+Na] ⁺ , 543 (80) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C ₂₈ H ₃₉ N ₄ 07 543.2813, found 543.2808.

Methyl-(2S,3/?,4S)-4-(4-(2,5,8,11 ,14-pentaoxaheptadec-16-yn-1 -yl)-1 H-1 ,2,3- triazol-1-yl)-1-acetyl-2-ethyl-3-methyl-1 ,2,3,4-tetrahydroquinoline-6-carboxylate (XS)

From 27 (100.0 mg, 0.32 mmol), 3,6,9,12,15-pentaoxaoctadec-1 ,17-diyne (256.0 mg, 0.95 mmol), 1 :1 H ₂ 0/f-Bu0H (4 ml_), CuS0 ₄ .5H ₂ 0 (16.0 mg, 0.06 mmol), and sodium ascorbate (63.0 mg, 0.22 mmol) and using method C, the product was obtained after flash chromatography (silica, 100% EtOAc) to give XS (79.2 mg, 43%); R _f = 0.29 (4% MeOH/CH ₂ CI ₂ ); [a] _D ²⁰ = +251.6 (c= 1.11 , CHCI ₃ ); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.92 (t, J = 7.2 Hz, 3H), 1.05 (d, J = 6.8 Hz, 3H), 1.22 (m, 1 H), 1.73 (m, 1 H), 2.38 (s, 3H), 2.43 (t, J= 2.4 Hz, 1 H), 2.59 (m, 1 H), 3.59-3.73 (m, 16H), 3.84 (s, 3H), 4.19 (d, J = 2.4 Hz, 2H), 4.70 (m, 3H), 5.59 (d, J = 11 .3 Hz, 1 H), 7.46 (m, 2H), 7.57 (d, J = 2.0 Hz, 1 H), 7.97 (d, J = 8.6, 2.0 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.7 (CH ₃ ), 16.1 (CH ₃ ), 18.4 (CH ₂ ), 24.0 (CH ₃ ), 40.5 (CH), 52.4 (CH ₃ ), 58.6 (CH ₂ ), 59.2 (CH), 63.1 (CH), 65.1 (CH ₂ ), 69.3 (CH ₂ ), 70.2 (CH ₂ ), 70.6 (CH ₂ ), 70.7 (3xCH ₂ ), 70.8 (2xCH ₂ ), 74.7 (CH), 79.8 (C), 120.9 (CH), 125.8 (CH), 127.0 (C), 127.6 (C), 130.1 (CH), 130.2 (CH), 140.6 (C), 146.6 (C), 165.9 (C), 170.1 (C); LRMS (ESI+) m/z (%) 609 (100) [/W+Na] ⁺ , 587 (50) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C _3O H4 ₃ 0 ₃ N 587.3075, found 587.3059.

Methyl-(2S,3/?,4S)-4-(4-(2,5,8,11 ,14,17-hexaoxaicos-19-yn-1 -yl)-1 H-1 ,2,3- triazol-1-yl)-1-acetyl-2-ethyl-3-methyl-1 ,2,3,4-tetrahydroquinoline-6-carboxylate (XT)

From 27 (100.0 mg, 0.32 mmol), 4,7,10,13,16,19-hexaoxadocosa-1 ,21-diyne (298.0 mg, 0.95 mmol), 1 :1 H ₂ 0/f-Bu0H (4 ml_), CuS04.5H ₂ 0 (16.0 mg, 0.06 mmol), and sodium ascorbate (63.0 mg, 0.22 mmol) and using method C, the product was obtained after flash chromatography (silica, 100% EtOAc, then 100 % CH ₂ CI ₂ and then gradient from 1 to 4 % MeOH in CH ₂ CI ₂ ) to give XT (90.0 mg, 45%); R _f = 0.29 (4% MeOH/CH ₂ CI ₂ ); ¹ H NMR (400 MHz, CDCI3) <5 ppm 0.92 (t, J= 7.2 Hz, 3H), 1.04 (d, J= 6.8 Hz, 3H), 1.22 (m, 1 H), 1.74 (m, 1 H), 2.38 (s, 3H), 2.43 (t, J = 2.4 Hz, 1 H), 2.59 (m, 1 H), 3.58-3.74 (m, 20H), 3.84 (s, 3H), 4.20 (d, J = 2.4 Hz, 2H), 4.70 (m, 3H), 5.58 (d, J = 11 .3 Hz, 1 H), 7.46 (m, 2H), 7.57 (d, J = 2.0 Hz, 1 H), 7.97 (d, J = 8.6, 2.0 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.7 (CH ₃ ), 16.1 (CH ₃ ), 18.4 (CH ₂ ), 24.0 (CH ₃ ), 40.4 (CH), 52.4 (CH ₃ ), 58.6 (CH ₂ ), 59.3 (CH), 63.2 (CH), 65.0 (CH ₂ ), 69.3 (CH ₂ ), 70.2 (CH ₂ ), 70.5 (3xCH ₂ ), 70.7 (3xCH ₂ ), 70.8 (2xCH ₂ ), 74.7 (CH), 79.8 (C), 121.2 (CH), 125.8 (CH), 127.0 (C), 127.6 (C), 130.1 (CH), 130.3 (CH), 140.5 (C), 146.4 (C), 165.9 (C), 170.2 (C); LRMS (ESI+) m/z (%) 653 (100) [/W+Na] ⁺ , 631 (55) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C32H47O9N4 631 .3338, found 631 .3327.

Dimethyl -4, 4'-(((ethane-1 ,2-diylbis(oxy))bis(methylene))bis(1 H-1 ,2,3-triazole- 4,1 -diyl))(2S,2'S,3/?,3'/?,4S,4'S)-bis(1 -acetyl-2-ethyl-3-methyl-1 ,2,3,4- tetrahydroquinoline-6-carboxylate) (XU) From 27 (69.6 mg, 0.22 mmol), 1 ,2-bis(prop-2-yn-1-yloxy)ethane (13.6 mg, 0.09 mmol), Amberlyst Cul.EtsN (40.0 mg), CH2CI2 (0.4 ml_), and using the method B with stirring for 72 h, the product was obtained after flash chromatography (silica gel, 100% EtOAc) to give XU (36.8 mg, 54%); R, = 0.37 (4% MeOH/CH ₂ CI ); [a] _D ²⁰ = +190.9 (c = 0.95, CHCI ₃ ); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.92 (t, J= 7.2 Hz, 6H), 1.03 (d, J= 6.9 Hz, 6H), 1.23 (m, 2H), 1 .74 (m, 2H), 2.38 (s, 6H), 2.61 (m, 2H), 3.70 (s, 4H), 3.82 (s, 6H), 4.67 (m, 6H), 5.59

(d, J = 11.2 Hz, 2H), 7.47 (m, 2H), 7.49 (s, 2H), 7.55 (d, J = 2.0 Hz, 2H), 7.96 (dd, J = 8.5, 2.0 Hz, 2H); ¹³ C NMR (125 MHz, CDCI ₃ ) <5 ppm 10.8 (2xCH ₃ ), 16.1 (2xCH ₃ ), 18.4 (2xCH ₂ ), 24.1 (2XCH ₃ ), 40.4 (2xCH), 52.4 (2xCH ₃ ), 59.3 (2xCH), 63.1 (2xCH), 65.1 (2xCH ₂ ), 70.1 (2XCH ₂ ), 121.1 (2xCH), 125.8 (2xCH), 127.0 (2xC), 127.6 (2xC), 130.1 (2xCH), 130.2 (2xCH), 140.6 (2xC), 146.4 (2xC), 165.9 (2xC), 170.1 (2xC); LRMS (ESI+) m/z (%) 793

(100) [MfNa] ⁺ ; HRMS (ESI+) m/z calc for C _4O H ₅ IN ₈ 0 ₈ [M+ H] ⁺ 771 .3824, found 771 .3824.

Dimethyl -4, 4'-((((oxybis(ethane-2,1 -diyl))bis(oxy))bis(methylene))bis(1 H-1 ,2,3- triazole-4,1 -diyl))(2S,2'S,3fl,3'fl,4S,4'S)-bis(1 -acetyl-2-ethyl-3-methyl-1 ,2,3,4- tetrahydroquinoline-6-carboxylate) (XV)

From 27 (100.0 mg, 0.32 mmol), 3-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)prop-1 - yne (23.0 mg, 0.13 mmol), Amberlyst Cul.EtsN (40.0 mg), CH2CI2 (1 ml_), and using the method B with stirring for 72 h, the product was obtained after flash chromatography (silica gel, 100% EtOAc, then 100 % CH ₂ CI ₂ and then gradient from 1 to 4 % MeOH in CH ₂ CI ₂ ) to give XV (39.3 mg, 38%); [a] _D ²⁰ = + 189.6 (c = 0.94, CHCI ₃ ); ¹ H NMR (400 MHz, CDCI ₃ ) d ppm 0.92 (t, J = 7.3 Hz, 6H), 1.03 (d, J = 6.8 Hz, 6H), 1.23 (m, 2H), 1.73 (m, 2H), 2.37 (s, 6H), 2.61 (m, 2H), 3.59-3.72 (m, 8H), 3.83 (s, 6H), 4.67 (m, 6H), 5.59 (d, J= 11 .3 Hz, 2H), 7.47 (m, 2H), 7.50 (s, 2H), 7.56 (d, J= 2.0 Hz, 2H), 7.96 (dd, J= 8.6, 2.0 Hz, 2H); ¹³ C NMR (125 MHz, CDCI ₃ ) <5 ppm 10.7 (2xCH ₃ ), 16.1 (2xCH ₃ ), 18.4 (2xCH ₂ ), 24.0 (2xCH ₃ ), 40.4 (2xCH), 52.4 (2XCH ₃ ), 59.4 (2xCH), 63.1 (2xCH), 65.1 (2xCH ₂ ), 70.1 (2xCH ₂ ), 70.6 (2XCH ₂ ), 121.1 (2xCH), 125.8 (2xCH), 127.0 (2xC), 127.5 (2xC), 130.1 (2xCH), 130.2 (2xCH), 140.6 (2xC), 146.5 (2xC), 165.9 (2xC), 170.1 (2xC); HRMS (ESI+) m/z calc for C ₄₂ H ₅₄ N ₈ 0 ₉ [M+ H] ⁺ 815.4087, found 815.4087.

Dimethyl -4, 4'-((2, 5, 8,11 -tetraoxadodecane-1 ,12-diyl)bis(1 H-1 ,2,3-triazole-4,1- diyl))(2S,2'S,3/?,3'/?,4S,4'S)-bis(1-acetyl-2-ethyl-3-methyl -1 ,2,3,4-tetrahydroquinoline- 6-carboxylate) (XW)

From 27 (111.0 mg, 0.35 mmol), 4,7,10,13-tetraoxahexadeca-1 ,15-diyne (31.6 mg, 0.14 mmol), Amberlyst Cul.Et ₈ N (40.0 mg), CH ₂ CI ₂ (1 ml_), and using the method B with stirring for 72 h, the product was obtained after flash chromatography (silica gel, 100% EtOAc, then 100 % CH ₂ CI ₂ and then gradient from 1 to 4 % MeOH in CH ₂ CI ₂ ) to give XW (45.1 mg, 38%); R _f = 0.24 (4% MeOH/CH ₂ CI ); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.92 (t, J = 7.2 Hz, 6H), 1.03 (d, J = 6.8 Hz, 6H), 1.22 (m, 2H), 1.73 (m, 2H), 2.37 (s, 6H), 2.60 (m, 2H), 3.61 -3.70 (m, 12H), 3.83 (s, 6H), 4.67 (m, 6H), 5.58 (d, J = 11 .3 Hz, 2H), 7.47 (m, 4H), 7.57 (d, J = 2.0 Hz, 2H), 7.96 (dd, J = 8.5, 2.0 Hz, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.7 (2XCH ₃ ), 16.1 (2XCH ₃ ), 18.4 (2xCH ₂ ), 24.0 (2xCH ₃ ), 40.4 (2xCH), 52.4 (2xCH ₃ ), 59.3 (2xCH), 63.2 (2xCH), 65.0 (2xCH ₂ ), 63.2 (2xCH ₂ ), 70.6 (4xCH ₂ ), 121.3 (2xCH), 125.8 (2xCH), 127.0 (2xC), 127.5 (2xC), 130.1 (2xCH), 130.2 (2xCH), 140.6 (2xC), 146.2 (2xC), 165.9 (2xC), 170.1 (2xC); LRMS (ESI+) m/z (%) 881 (100) [/W+Na] ⁺ , 859 (85) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C ₄₄ H ₅₉ N ₈ OI _O 859.4349, found 859.4344. Dimethyl -4, 4'-((2, 5, 8,11 ,14-pentaoxapentadecane-1 ,15-diyl)bis(1 H-1 ,2,3- triazole-4,1 -diyl))(2S,2'S,3fl,3'fl,4S,4'S)-bis(1 -acetyl-2-ethyl-3-methyl-1 ,2,3,4- tetrahydroquinoline-6-carboxylate) (XX)

From 27 (51.6 mg, 0.19 mmol), 3,6,9,12,15-pentaoxaoctadec-1 ,17-diyne (20.5 mg, 0.08 mmol), Amberlyst Cul.EtsN (33.0 mg), CH ₂ CI ₂ (0.4 ml_), and using the method B with stirring for 72 h, the product was obtained after flash chromatography (silica gel, 100% EtOAc, then 100 % CH ₂ CI ₂ and then gradient from 1 to 4 % MeOH in CH ₂ CI ₂ ) to give a mixture of monomer and dimer. This mixture was purified by flash silica chromatography (silica gel, 6% MeOH/EtOAc) to give the desired product XX (42.7 mg, 62%); R _f = 0.19 (6% MeOH/EtOAc); [a] _D ²⁰ = +130.3 (c = 0.58, CHCI ₃ ); IR v _max (thin film, CH ₂ CI ₂ ) 3420, 2962, 2875, 1717, 1661 , 1610, 1259 cm ¹ ; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.92 (t, J = 7.2 Hz, 6H), 1.04 (d, J = 6.8 Hz, 6H), 1.23 (m, 2H), 1.72 (m, 2H), 2.38 (s, 6H), 2.59 (m, 2H), 3.57- 3.72 (m, 16H), 3.83 (s, 6H), 4.68 (s, 6H), 5.58 (d, J = 11 .2 Hz, 2H), 7.46 (s, 4H), 7.57 (d, J

= 2.0 Hz, 2H), 7.96 (dd, J = 8.5, 2.0 Hz, 2H); ¹³ C NMR (125 MHz, CDCI ₃ ) <5 10.8 (2xCH ₃ ), 16.2 (2XCH ₃ ), 18.5 (2xCH ₂ ), 24.1 (2xCH ₃ ), 40.5 (2xCH), 52.5 (2xCH ₃ ), 59.3 (2xCH), 63.1 (2xCH), 65.2 (2XCH ₂ ), 70.2 (2xCH ₂ ), 70.8 (6xCH ₂ ), 121.1 (2xCH), 125.8 (2xCH), 127.1 (2xC), 127.6 (2xC), 130.2 (2xCH), 130.2 (2xCH), 140.7 (2xC), 146.7 (2xC), 165.9 (2xC), 170.1 (2xC); LRMS (ESI+) m/z (%) 925 (100) [/W+Na] ⁺ ; HRMS (ESI+) m/z calc for

C ₄₆ H ₆₃ N ₈ 0II [M+ H] ⁺ , 903.4611 , found 903.4611 . Dimethyl -4, 4'-((2, 5, 8,11 ,14,17-hexaoxaoctadecane-1 ,18-diyl)bis(1 H-1 ,2,3- triazole-4,1 -diyl))(2S,2'S,3fl,3'fl,4S,4'S)-bis(1 -acetyl-2-ethyl-3-methyl-1 ,2,3,4- tetrahydroquinoline-6-carboxylate) (XY) From 27 (71.9 mg, 0.23 mmol), 4,7,10,13,16,19-hexaoxadocosa-1 ,21 -diyne (28.8 mg, 0.09 mmol), Amberlyst Cul.EtsN (40.0 mg), CH2CI2 (0.4 ml_), and using the method B with stirring for 72 h, the product was obtained after flash chromatography (silica gel, 100% EtOAc, then 100 % CH2CI2 and then gradient from 1 to 4 % MeOH in CH2CI2) to give a mixture of monomer and dimer. This mixture was purified by flash silica chromatography (silica gel, 6 to 8% MeOH/EtOAc) to give the desired product XY (29.5 mg, 26%); R _f = 0.14 (4% MeOH/EtOAc); [a] _D ²⁰ = +130.3 (c = 0.58, CHCI ₃ ); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.92 (t, J = 7.2 Hz, 6H), 1.04 (d, J = 6.8 Hz, 6H), 1.22 (m, 2H), 1.74 (m, 2H), 2.38 (s, 6H), 2.60 (m, 2H), 3.57-3.72 (m, 20H), 3.83 (s, 6H), 4.69 (s, 6H), 5.58 (d, J = 11 .2 Hz, 2H), 7.47 (s, 4H), 7.57 (d, J= 2.0 Hz, 2H), 7.97 (dd, J= 8.5, 2.0 Hz, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.7 (2xCH ₃ ), 16.1 (2xCH ₃ ), 18.4 (2xCH ₂ ), 24.0 (2xCH ₃ ), 40.5 (2xCH), 52.4 (2xCH ₃ ),

59.3 (2xCH), 63.1 (2xCH), 65.2 (2xCH ₂ ), 70.2 (2xCH ₂ ), 70.6 (2xCH ₂ ), 70.7 (4xCH ₂ ), 121 .0 (2xCH), 125.8 (2xCH), 127.1 (2xC), 127.6 (2xC), 130.2 (2xCH), 130.2 (2xCH), 140.6 (2xC), 146.6 (2xC), 165.9 (2xC), 170.1 (2xC); LRMS (ESI+) m/z (%) 969 (100) [/W+Na] ⁺ ; HRMS (ESI+) m/z calc for C ₄ 8H ₆ 7N ₈ Oi2 [M+ H] ⁺ 947.4873, found 947.4873.

Methyl-(2S,3/?,4S)-1-acetyl-4-(4-(14-(2-(2,6-dioxopiperid in-3-yl)-1- oxoisoindolin-4-yl)-2,5,8,11-tetraoxatetradec-13-yn-1-yl)-1H -1,2,3-triazol-1-yl)-2- ethyl-3-methyl-1 ,2,3,4-tetrahydroquinoline-6-carboxylate (YF)

To a solution of 3-(4-iodo-1-oxoisoindolin-2-yl)piperidine-2,6-dione ^[41 (11 mg, 0.02 mmol) and XR (9.5 mg, 0.257 mmol) in dry DMF (1 mL) were added Pd(PPh ₃ )2Cl2 (1 .4 mg, 0.00197 mmol) and Cul (0.2 mg, 0.000985 mmol) in triethylamine (40 mI_). The mixture was degazed by argon and purged many times. The reaction was stirring at 80°C for 6 h under Argon. After cooling at room temperature, the mixture was concentrated in vacuo. After flash chromatography (silica gel, 100% EtOAC, then EtOAc/MeOH (1 -4%)), the product was isolated as a yellow oil (6.3 mg, 40%).

R _f = 0.30 (4% MeOH/CH ₂ CI ); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.92 (t, J= 7.2 Hz, 3H), 1 .04 (m, 3H), 1 .23 (m, 1 H), 1 .74 (m, 1 H), 2.24 (m, 1 H), 2.37 (s, 3H), 2.44 (m, 1 H), 2.62 (m, 1 H), 2.84 (m, 1 H), 2.92 (m, 1 H), 3.55-3.80 (m, 12H), 3.84 (s, 3H), 4.40 (d, J = 16.7 Hz, 1 H), 4.47 (s, 2H), 4.53 (dd, J= 16.7, 6.0 Hz, 1 H), 4.69 (bs, 2H), 4.71 (m, 1 H), 5.23 (m, 1 H), 5.59 (m, 1 H), 7.46 (t, J= 7.6 Hz, 1 H), 7.51 (m, 2H), 7.57 (d, J= 1 .9 Hz, 1 H), 7.61 (d, J= 7.6 Hz, 1 H), 7.84 (d, J= 7.6 Hz, 1 H), 7.96 (dd, J= 7.6, 1.9 Hz, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.8 (CH ₃ ), 16.2 (CH ₃ ), 18.5 (CH ₂ ), 23.6 (CH ₂ ), 24.1 (CH ₃ ), 31.8 (CH ₂ ), 40.5 (CH), 47.3 (CH ₂ ), 52.1 (CH), 52.5 (CH ₃ ), 59.4 (CH ₂ ), 59.6 (CH), 63.2 (CH), 65.1 (CH ₂ ), 69.5 (CH ₂ ), 70.1 (CH ₂ ), 70.5 (2XCH ₂ ), 70.6 (CH ₂ ), 70.7 (CH ₂ ), 82.1 (C), 90.8 (C), 1 18.4 (C), 120.1 (CH), 124.4 (CH), 125.9 (CH), 127.0 (C), 127.6 (C), 128.7 (CH), 130.1 (CH), 130.2 (CH), 132.0 (CH), 135.0 (CH), 140.6 (C), 144.1 (C), 146.5 (C), 166.0 (C), 169.0 (C), 169.7 (C), 170.2 (C), 171.3 (C); LRMS (ESI+) m/z (%) 807 (100) [/W+Na] ⁺ , 785 (48) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C41H49N6O10 [M+ H] ⁺ 785.3505, found 785.3502. Methyl-(2S,3/?,4S)-1-acetyl-4-(4-((2-((3-(2-(2,6-dioxopipend in-3-yl)-1- oxoisoindolin-4-yl)prop-2-yn-1-yl)oxy)ethoxy)methyl)-1 H-1 ,2,3-triazol-1-yl)-2-ethyl-3- methyl-1 ,2,3,4-tetrahydroquinoline-6-carboxylate (YE)

Using the same protocol as YF, 15.0 mg of compound YE was obtained (52%);

R _f = 0.30 (4% MeOH/CH ₂ CI ); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.92 (t, J= 7.2 Hz, 3H), 1 .04 (m, 3H), 1 .22 (m, 1 H), 1 .72 (m, 1 H), 2.23 (m, 1 H), 2.37 (s, 3H), 2.48 (m, 1 H), 2.57 (m, 1 H), 2.85 (m, 1 H), 2.95 (m, 1 H), 3.65-3.79 (m, 4H), 3.83 (s, 3H), 4.41 (dd, J = 16.8, 3.4 Hz, 1 H), 4.43 (s, 2H), 4.54 (dd, J = 16.8, 6.1 Hz, 1 H), 4.67-4.72 (m, 3H), 5.24 (dt, J = 13.3, 5.3 Hz, 1 H), 5.59 (m, 1 H), 7.44 (s, 1 H), 7.46 (t, J = 7.3 Hz, 1 H), 7.48 (m, 1 H), 7.57 (s, 1 H), 7.61 (dd, J = 7.6, 1 .0 Hz, 1 H), 7.85 (d, J = 7.6 Hz, 1 H), 7.96 (m, 1 H) ); ¹³ C NMR (100 MHz, CDCh) <5 ppm 10.6 (CH ₃ ), 15.9 (CH ₃ ), 18.2 (CH ₂ ), 23.4 (CH ₂ ), 23.8 (CH ₃ ), 31 .6 (CH ₂ ), 40.4 (CH), 47.1 (CH ₂ ), 51 .9 (CH), 52.3 (CH ₃ ), 59.1 (CH ₂ ), 59.3 (CH), 62.9 (CH), 64.9 (CH ₂ ), 69.2 (CH ₂ ), 69.7 (CH ₂ ), 82.0 (C), 90.4 (C), 118.1 (C), 120.7 (CH), 124.7 (CH), 125.7 (CH), 126.8

(C), 127.4 (C), 128.4 (CH), 129.9 (CH), 130.0 (CH), 131.8 (CH), 134.8 (CH), 140.4 (C), 143.9 (C), 146.2 (C), 165.8 (C), 168.8 (C), 169.5 (C), 170.0 (C), 171.1 (C); LRMS (ESI+) m/z (%) 719 (100) [/W+Na] ⁺ , 697 (90) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C ₃₇ H _4i N ₆ 0 ₈ [M+ H] ⁺ 697.2980, found 697.2978. Methyl-(2S,3/?,4S)-1-acetyl-4-(4-(20-(2-(2,6-dioxopipendin-3 -yl)-1- oxoisoindolin-4-yl)-2,5,8,11,14,17-hexaoxaicos-19-yn-1-yl)-1 H-1,2,3-triazol-1-yl)-2- ethyl-3-methyl-1 ,2,3,4-tetrahydroquinoline-6-carboxylate (YG)

Using the same protocol as YF, 15.0 mg of compound Y was obtained (59%);R _f = 0.28 (4% MeOH/CH ₂ CI ); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.92 (t, J= 7.2 Hz, 3H), 1.04 (d ; J= 6.8 Hz, 3H), 1.22 (m, 1H), 1.73 (m, 1H), 2.23 (m, 1H), 2.38 (s, 3H), 2.48 (m, 1H), 2.65 (m, 1 H), 2.84 (m, 1H), 2.92 (m, 1H), 3.59-3.80 (m, 20H), 3.84 (s, 3H), 4.44 (m, 1H), 4.48 (s, 2H), 4.55 (dd, J= 16.9, 2.5 Hz, 1H), 4.71 (bs, 3H), 5.22 (m, 1H), 5.60 (d, J= 11.2 Hz, 1H), 7.46 (t, = 7.7 Hz, 1H), 7.51 (bs, 2H), 7.57 (d, J= 1.1 Hz, 1H), 7.62 (d, J=7.7 Hz, 1 H), 7.84 (d, J= 7.7, 1.1 Hz, 1H), 7.96 (d, J= 7.7 Hz, 1H); ¹³ C NMR (100 MHz, CDCI ₃ ) d ppm 10.8 (CH ₃ ), 16.2 (CH ₃ ), 18.5 (CH ₂ ), 23.6 (CH ₂ ), 24.1 (CH ₃ ), 31.8 (CH ₂ ), 40.4 (CH), 47.5 (CH ₂ ), 52.2 (CH), 52.5 (CH ₃ ), 59.4 (CH ₂ ), 59.6 (CH), 63.1 (CH), 64.9 (CH ₂ ), 69.3 (CH ₂ ), 69.8 (CH ₂ ), 70.3 (6XCH ₂ ), 70.5 (2xCH ₂ ), 82.2 (C), 90.7 (C), 118.3 (C), 121.3 (CH), 124.4 (CH), 125.9 (CH), 127.0 (C), 127.5 (C), 128.6 (CH), 130.1 (CH), 130.2 (CH), 132.0 (CH), 134.8 (CH), 140.7(C), 144.1 (C), 146.2(C), 166.0(C), 169.0 (C), 169.8(C), 170.2(C), 171.4(C); LRMS (ESI+) m/z (%) 895 (100) [/W+Na] ⁺ , 873 (48) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C ₅ H N ₆ OI ₂ [M+ H] ⁺ 873.4029, found 873.4029. 2S,3/?,4S)-4-azido-2-ethyl-3-methyl-6-(trifluoromethyl)-1 ,2,3,4- tetrahydroquinoline (YA)

To a solution of 4(trifluoromethyl)aniline (506.7 mg, 3.145 mmol) and propionaldehyde (41.52g, 77.58 mmol) in CH ₃ CN (31 mL) at -20°C was added D-proline (109.4 mg, 0.955 mmol). The reaction mixture was stirred for 20 min at -20°C and kept at -20°C for two days. TMSN3 (0.58 mL, 4.027 mmol) and BF ₃ Et ₂ 0 (0.28 mL, 2.201 mmol) were added at rt and the reaction mixture was stirred at this temperature for 30 min. After the reaction was complete, it was quenched by NaOH (1 N) until neutralization, extracted with EtOAc, dried by MgSC>4, filtered off and concentrated under vacuum. After flash chromatoghraphy (silica gel, 1% EtOAc/5% DCIW 86% cyclohexane), the product YA was isolated (710.9 mg, 80%); R _f = 0.32 (2% EtOAc/cyclohexane); [a] _D ²⁰ = -207.1 (c=0.78, CHCI3); IR vm _ax (thin film, CH2CI2) 2970, 2090, 1622, 1520, 1330, 1109 cm ¹ ; ¹ H NMR (400 MHz, CDCI3) <5 ppm 0.75 (d, J = 7.1 Hz, 3H), 1.02 (t, J = 7.5 Hz, 3H), 1.53-1.60 (m, 2H), 1 .97 (m, 1 H), 3.38 (td, J = 7.2, 2.8 Hz, 1 H), 4.20 (s, 1 H), 4.24 (d, J = 2.8 Hz, 1 H), 6.56 (d, J = 8.1 Hz, 1 H), 7.30-7.35 (m, 2H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.4 (CH ₃ ), 10.5 (CH ₃ ), 25.3 (CH ₂ ), 33.2 (CH), 51.0 (CH), 63.7 (CH), 113.9 (CH), 115.0 (C), 118.4 (C, q, J = 32.8 Hz), 124.9 (C, q, J= 270.5 Hz), 126.9 (C, q, J= 3.5 Hz), 128.6 (C, q, J= 7.5 Hz), 146.8 (C); LRMS (ESI+) m/z (%) 304 (100), 278 (10) [/W+Na] ⁺ ; HRMS (ESI-) m/z calc for C13H15 F3N4CI [/W+CI]-319.0942, found 319.0929.

1-((2S,3/?,4S)-4-azido-2-ethyl-3-methyl-6-(tnfluoromethyl )-3,4- dihydroquinolin-1 (2H)-yl)ethan-1 -one (YB)

To a solution of YA (638.6 mg, 2.21 mmol) and pyridine (0.53 mL, 6.63 mmol) in CH2CI2 (5 mL) was added acetyl chloride (0.32 mL, 4.42 mmol) at 0°C. The reaction mixture was stirred at 0°C to rt for 2h30 and then stirred at rt overnight. The reaction was concentrated under vacuum. After flash chromatography (silica gel, 20% EtOAc/cyclohexane), the product YB was isolated (350.1 mg, 48.5%); R _f = 0.12 (10% EtOA c/cyclohexane); [a] _D ²⁰ = +145.6 (c=1 , CHCI ₃ ); IR v _max (thin film, CH ₂ CI ): 3478, 3055, 2969, 2486, 2097, 1666, 1331 , 1169, 737 cm ¹ ; ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.88 (t, J = 7.2 Hz, 3H), 1.09 (m, 1 H), 1.25 (d, J = 6.9 Hz, 3H), 1.64 (m, 1 H), 2.30 (s, 3H), 2.34 (m, 1 H), 4.11 (d, J = 10.4 Hz, 1 H), 4.62 (bs, 1 H), 7.49 (m, 1 H), 7.56 (dd, J = 8.5, 2.1 Hz, 1 H), 7.72 (bs, 1 H); ¹³ C NMR (100 MHz, CDCI ₃ ) <5 ppm 10.6 (CH ₃ ), 16.7 (CH ₃ ),18.3 (CH ₃ ), 23.6 (CH ₂ ), 38.8 (CH), 59.1 (CH), 63.3 (CH), 119.7 (CH), 122.4 (C), 125.1 (C), 125.4 (C, q, J = 125.4 Hz), 125.9 (C), 126.2 (C, q, J = 126.21 Hz), 127.3 (C, q, J = 127.5 Hz), 128.6 (C), 139.8 (C), 169.9 (C); LRMS (ESI+) m/z (%) 242 (100) 327 (40) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for CI ₅ HI ₈ N ₄ OF ₃ 327.1433 found 327.1425.

1 -((2S,3fl,4S)-2-Ethyl-3-methyl-4-(4-phenyl-1 H-1 ,2,3-triazol-1 -yl)-6- (trifluoromethyl)-3,4-dihydroquinolin-1(2H)-yl)ethan-1-one (YC)

From YB (55.0 mg, 0.17 mmol), phenylacetylene (37 mI_, 0.34 mmol), Amberlyst Cul.Et ₃ N (20.0 mg), CH ₂ CI ₂ (0.5 mL), and using the method B with stirring for 72 h, the product was obtained after flash chromatography (silica, 20% to 40% EtOAc/cyclohexane) to give YC (67.7 mg, 93.5%); R _f = 0.26 (20% EtOAc/cyclohexane); ¹ H NMR (400 MHz, CDCIs) <5 ppm 0.97 (t, J = 7.3 Hz, 3H), 1 .09 (d, J = 6.9 Hz, 3H), 1 .26 (m, 1 H), 1 .78 (m, 1 H), 2.40 (s, 3H), 2.67 (m, 1 H), 4.65 (m, 1 H), 5.66 (d, J = 11 .5 Hz, 1 H), 7.22 (s, 1 H), 7.35 (d, J = 7.5 Hz, 1 H), 7.39-7.45 (m, 3H), 7.56-7.60 (m, 2H), 7.83 (d, J= 7.5 Hz, 2H);; LRMS (ESI+) m/z (%) 429 (100) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C ₂₃ H ₂₄ N ₄ OF ₃ 429.1902, found 429.1898.

1 -((2S,3fl,4S)-2-ethyl-3-methyl-4-(4-pentyl-1 H-1 ,2,3-triazol-1 -yl)-6- (trifluoromethyl)-3,4-dihydroquinolin-1(2H)-yl)ethan-1-one (YD) From YB (100.0 mg, 0.31 mmol), 1 -heptyne (59 mg, 0.61 mmol), Amberlyst Cul.EtsN (40.0 mg), CH2CI2 (1 ml_), and using the method B with stirring for 48 h, the product was obtained after flash chromatography (silica, 20% to 30% EtOAc/cyclohexane) to give YD (107.5 mg, 83%); R _f = 0.38 (30% EtOAc/cyclohexane); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.87 (t, J = 6.8 Hz, 3H), 0.95 (t, J = 7.2 Hz, 3H), 1 .04 (d, J = 6.8 Hz, 3H), 1 .2-1 .38 (m, 5H), 1.61-1.71 (m, 3H), 1.75 (m, 1 H), 2.38 (s, 3H), 2.59 (m, 1 H), 2.71 (t J = 7.6 Hz, 2H), 4.63 (bs, 1 H), 5.57 (d, J = 11.3 Hz, 1 H), 7.12 (m, 1 H), 7.15 (m, 1 H), 7.56 (d, J = 8.0 Hz, 1 H);; LRMS (ESI+) m/z (%) 423 (100) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C22H30N4OF3423.2372, found 423.2374.

N-(4-((2S,3/?,4S)-1-acetyl-4-azido-2-ethyl-3-methyl-1,2,3 ,4-tetrahydroquinolin- 6-yl)phenyl)-N-hydroxyoctanediamide (YH)

R r= 0.22 (5% MeOH/CH ₂ CI ₂ ); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.85 (d, J= 7.3 Hz, 3H), 1.11 (m, 1 H), 1 .21 (d, J = 6.9 Hz, 3H), 1 .27-1 .46 (m, 5H), 1 .64 (m, 2H), 1 .71 (m, 2H), 2.09 (t, J= 7.4 Hz, 2H), 2.29 (s, 3H), 2.31 (m, 1 H), 2.39 (t, J = 7.4 Hz, 2H), 4.32 (d, J= 9.5 Hz, 1 H), 4.64 (m, 1 H), 7.23 (d, J = 7.9 Hz, 1 H), 7.44 (m, 1 H), 7.60 (d, J = 8.2 Hz, 2H), 7.66 (m, 1 H), 7.70 (d, J = 8.2 Hz, 2H); ¹³ C NMR (100 MHz, CD ₃ OD) d ppm 11.0 (CH ₃ ), 16.9 (CH ₃ ),19.5 (CH), 21.4 (CH ₃ ), 26.7 (CH ₂ ), 26.8 (CH ₂ ), 29.9 (CH ₂ ), 30.0 (CH ₂ ), 34.8 (CH ₂ ), 38.0 (CH ₂ ), 40.9 (CH), 60.4 (CH), 65.0 (CH), 121.7 (CH), 127.1 (2xCH), 127.7 (CH), 128.2 (CH), 129.9 (2xCH), 130.6 (C), 136.7 (C), 139.8 (C), 141.9 (C), 143.6 (C), 172.8 (C), 173.1 (C), 174.8 (C); LRMS (ESI+) m/z (%) 521 (30) [M+ H] ⁺ , 543 (100) [/W+Na] ⁺ ; HRMS (ESI+) m/z calc for C23H37N6O4 521 .2863, found 521 .2863.

Methyl-(2S,3/?,4S)-1-acetyl-2-ethyl-4-(6-(hydroxymethyl)- 5,5a,6,6a,7,8- hexahydrocyclopropa[5,6]cycloocta[1 ,2-d][1 ,2,3]triazol-1 (4H)-yl)-3-methyl-1 ,2,3,4- tetrahydroquinoline-6-carboxylate (Yl)

Compound 26 (21 mg, 0.06 mmol) and bicyclo[6.1 0]non-4-yn-9-ylmethanol (BCN, 10 mg, 0.06 mmol) were dissolved in CHCl ₃ (1 mL) and the reaction was stirred for 2 h at room temperature. The solvent was removed in vacuo and the product was obtained after flash chromatography (silica, 4% MeOH/CH ₂ CI ₂ ) to give Yl (13 mg, 42%);

R _f = 0.33 (4% MeOH/CH ₂ CI ); ¹ H NMR (400 MHz, CDCI ₃ ) <5 ppm 0.93 (d, J= 7.2 Hz, 3H), 0.96-1.34 (m, 9H), 1.44-1.59 (m, 2H), 1.67-1.95 (m, 3H), 2.26 (m, 1 H), 2.36 (s, 3H), 2.48-2.79 (m, 2H), 2.92 (m, 1 H), 3.11 (m, 1 H), 3.61 (m, 1 H), 3.72 (m, 1 H), 3.82 (s, 3H), 4.81 (m, 1 H), 5.59 (m, 1 H), 7.42 (m, 1 H), 7.48 (m, 1 H), 7.96 (m, 1 H); ); LRMS (ESI+) m/z (%) 489 (88) [MfNa] ⁺ , 467 (100) [M+ H] ⁺ ; HRMS (ESI+) m/z calc for C _M H ₃₅ N ₄ O ₄ [M+H] ⁺ 467.2653, found 467.2646.

References

[1] Hatano, M.; Yamashita, K.; Ishihara, K. Org. Let. 2015, 17 , 2412-2415

[2] Allais, F.; Ducrot, P.-H. Synthesis 2010, 1649-1653

[3] Girard, C.; Onen, E.; Aufort, M.; Beauviere, S.; Samson, E.; Herscovici, J. Org. Let. 2006, 8, 1689-1692.

[4] Chong, Q.; Hu, Y. Zhou, B.; Fernandez-Salas, E.; Yang, C.-Y.; Liu, L.; McEachern, D. et al. J. Med. Chem. 2018, 67,6685-6704. In vitro experimentations

Evaluation of cytotoxicity and cytokine expression after iBET/LPS treatment

J774 mouse macrophages were cultured in DMEM 10% Fetal Calf Serum (FCS) 1 % penicillin and streptomycin at 37°C 5% C02. 250,000 cells/ ml. Cells were then seeded at 250,000 cells/ml and treated with 1 mM, 10 mM and 100 pM for each iBET and, then, after 30 min, with 100gg/ml LPS (lipopolysaccharides, Cell Signaling Technology ref: 14011 S) to induce inflammatory response. After 24h, cytotoxicity was evaluated using CytoTox-ONE™ Flomogeneous Membrane Integrity Assay and/or CellTiter 96 Non-radioactive Cell Proliferation Assay (Promega) according to manufacturer’s instructions. IL6 and TNFa secretion were assessed using respectively

IL-6 Mouse ELISA assay and TNFa Mouse ELISA assay (Invitrogen) according to manufacturer’s instructions.

Results are given on figures 1 (J774) and 2 to 5 (IL-6 and TNFa).

In vitro evaluation of bromodomain inhibitors by Homogeneous Time Resolved Fluorescence

HTRF reagents and buffers were purchased from Cisbio Bioassays. The assay used a terbium (III) cryptate donor reagent conjugated to an anti-GST antibody (MAb anti-GST- Tb; GSTTLA), a streptavidin-conjugated acceptor reagent (streptavidin-d2) and Cisbio proprietary buffers (EPIgeneous Binding Domain Diluent and Detection buffer, respectively). GST-tagged bromodomains (BDs) were expressed in E. coli and purified using standard procedures. Incubation of GST-tagged BDs with biotinylated acetylated H4 peptide (H4K5acK8acK12acK16ac, here named H4ac4) brings the donor and acceptor into close proximity and allows for a FRET reaction. GST-tagged proteins in 25 mM Hepes pH 7.5, 150 mM NaCI, 0.5 mM DTT were assayed at a final concentration of 5 nM. Biotinylated H4ac4 peptides were used at a final concentration of 50, 600 nM in assays involving Brd4 BD1 , Brd4 BD2, respectively. The antibody-conjugated donor was used at 0.5 nM and the streptavidin-conjugated acceptor was used at 1/8 of the H4ac4 peptide concentration. Inhibitors were tested by performing an eleven-point dilution series with a maximal final concentration of 20 mM. These concentrations allowed a fixed DMSO concentration at 0.2 %, critical for a Z’ factor ³ 0.8. Components were incubated at 4 °C for 4 h (BD1) or for 24 h (BD2). Experiments were performed in 384-well white plates (Greiner ref. 781080) and analyzed in a ClarioStar plate reader (BMG LABTECH, excitation at 330 nm and emission at 620 and 665 nm, corresponding to the donor and acceptor emission peaks, respectively; the 665/620 ratio is used to calculate the specific HTRF signal) with an integration delay of 60 ps and an integration time of 400 ps.

HTRF Results (IC50 sur BD1 et BD2) Some of the HTRF results are illustrated on Figure 6.

89

Evaluation of cytotoxicity and anti-inflammatory activity of BET inhibitor compounds

Experimental procedure: J774 mouse macrophages were cultured in DMEM 10% Fetal Calf Serum (FCS) 1% penicillin and streptomycin at 37°C 5% CO2. Cells were seeded at 250,000 cells/mL and treated with 1 mM, 10 mM and 100 pM for each iBET and, after 30 min, with 100 pg/mL LPS (lipopolysaccharides, Cell Signaling Technology) to induce inflammatory response. After 24h, cytotoxicity was evaluated using the Cell Proliferation Kit I (Roche) according to manufacturer’s instructions. IC50 were calculated using GraphPad (Prism) using non linear regression curve fit. IL6 and TNFD Dsecretion was assessed using respectively IL-6 Mouse and TNFa Mouse ELISA assay (Invitrogen) on cell culture supernatants according to manufacturer’s instructions. Normalized values were combined to calculate anti-inflammatory (Al) score.

As shown in Figure 7, compounds show a wide variety of Al score and cytotoxicity. Interestingly, among all, some show a good ratio between biological activity (Al score) and low level of cell toxicity and therefore have a good potential for clinical transfer.

Impact of “lead” BET inhibitors compounds on gene expression profiles

Experimental procedure: RNA was extracted from 5 million of LPS-stimulated J774 mouse macrophages cells treated for 6h with DMSO (negative control), reference iBETs (JQ1 and iBET726, 1 pM) and lead compounds (MLE83 and JUP102, 10 pM) using Trizol (Invitrogen) according to manufacturer’s instructions. RNA sequencing was subcontracted to BGI Genomics (DNBSEQ™ Sequencing Technology). FASTQ sequencing files were aligned to mouse genome (GRCm39 assembly) using STAR (v2.7.1a). Bam files were counted using HTSeq framework (vO.11.2). Reads normalization and differential analysis was performed suing SARTools and DESeq2 (v1.22.2) R packages. Genes were considered as differentially expressed when |log2FC|>1 AND p value < 0.05 between control and treated conditions. Gene Set Enrichment Analysis was performed using GSEA v4.2.1 software against the hallmark v7.5 gene sets from the Broad Institute Molecular Signature Database. A detailed description of GSEA methodology and interpretation is provided online (http://www.broadinstitute.org/gsea/doc/GSEAUserGui- deFrame.html). As shown in Figure 8, for the two molecules with the best inflammatory activity/toxicity ratio, the pan-iBET MLE83 and the BD2-selective compound JUP102, we performed RNAseq analyses to precisely characterize the impact of these different compounds, compared to reference iBETs (JQ1 and iBET726) on the gene expression profiles of LPS-treated J774 (murine macrophage line) cells. This analysis showed that the impact of the new compounds on the gene expression profiles was less important, although they preserved an anti-inflammatory activity comparable to that of the commercial compounds. Interestingly, this appears to be independent of the selectivity of the molecule: the pan iBET compound, MLE83 impacting nearly 10 times fewer genes than the BD2-selective compound, JUP102.

GSEA analysis revealed that the limited number of genes impacted by JUP102 and MLE83 are still able to mediate iBET -associated cellular responses, i.e. reduces inflammatory response and MYC signaling. These data strongly suggest that these compounds, having an impact on a limited number of target genes, should have a lower toxicity and be associated with fewer side effects in vivo. iBET-726 (reference) has the following structure: