Novel Bifunctional Molecules For Targeted Protein Degradation

Abstract

The present disclosure relates to a novel class of bifunctional molecules that are useful in a targeted or selective degradation of a protein.

Claims

1. A bifunctional molecule comprising the general formula:
TBL-L-Z wherein TBL is a target protein binding ligand; L is a linker; and Z comprises a structure according to formula (I): ##STR00463## wherein R.sup.1 is selected from C.sub.1 to C.sub.6 alkyl, benzyl, substituted benzyl, carbocyclyl, substituted carbocyclyl, heterocyclyl and substituted heterocyclyl, optionally wherein the C.sub.1 to C.sub.6 alkyl is substituted with one or more heteroatoms selected from halo, N, O and S and/or is substituted with a carbocyclic or heterocyclic group; A is absent or is CR.sup.2R.sup.2; B is selected from aryl, heteroaryl, substituted aryl and substituted heteroaryl; R.sup.2 and R.sup.2 are each independently selected from H and C.sub.1 to C.sub.6 alkyl, optionally wherein the C.sub.1 to C.sub.6 alkyl is substituted with one or more heteroatoms selected from N, O or S, or wherein R.sup.2 and R.sup.2 together form a 3-, 4-, 5- or 6-membered carbocyclic or heterocyclic ring; R.sup.3 is selected from C.sub.1 to C.sub.6 alkyl, aryl, heteroaryl, substituted aryl, substituted heteroaryl, carbocyclyl, substituted carbocyclyl, heterocyclyl and substituted heterocyclyl, optionally wherein the C.sub.1 to C.sub.6 alkyl is substituted with one or more heteroatoms selected from halo, N, O and S and/or is substituted with a carbocyclic or heterocyclic group; R.sup.4 is H, C.sub.1 to C.sub.6 alkyl, optionally wherein the C.sub.1 to C.sub.6 alkyl is substituted with one or more heteroatoms selected from N, O or S; or wherein R.sup.1 and R.sup.4 together form a 5-, 6-, or 7-membered heterocyclic ring; or wherein when A is CR.sup.2R.sup.2: R.sup.1 and R.sup.2 together form a 5-, 6-, or 7-membered heterocyclic ring; or R.sup.2 and R.sup.4 together form a 5-, 6-, or 7-membered heterocyclic or carbocyclic ring; and L shows the point of attachment of the linker.

2. A bifunctional molecule according to claim 1, wherein: (i) when R.sup.1 and R.sup.4 together form a 5-, 6-, or 7-membered heterocyclic ring, Z is represented by formula (Ia): ##STR00464## wherein A, B, R.sup.3 and L are as defined for formula (I); and n is 1, 2 or 3; W is selected from CR.sup.W1R.sup.W2, O, NR.sup.W3 and S; R.sup.W1, R.sup.W2 and R.sup.W3 are each independently selected from H and C.sub.1 to C.sub.6 alkyl; and wherein when n is 2 or 3, each W is independently selected from CR.sup.W1R.sup.W2, O, NR.sup.W3, and S; (ii) when R.sup.1 and R.sup.2 together form a 5-, 6-, or 7-membered heterocyclic ring, Z is represented as formula (Ib): ##STR00465## Wherein B, R.sup.2, R.sup.3, R.sup.4 and L are as defined for formula (I); m is 3, 4 or 5; each T is independently selected from CR.sup.T1R.sup.T2, O, NR.sup.T3 and S; and R.sup.T1, R.sup.T2 and R.sup.T3 are each independently selected from H and C.sub.1 to C.sub.6 alkyl; or (iii) when R.sup.2 and R.sup.4 together form a 5-, 6-, or 7-membered heterocyclic or carbocyclic ring, Z is represented as formula (Ic): ##STR00466## Wherein B, R.sup.1, R.sup.2, R.sup.3 and L are as defined for formula (I); p is 2, 3 or 4; and each U is independently selected from CR.sup.U1R.sup.U2, O, NR.sup.U3 and S; and R.sup.U1, R.sup.U2 and R.sup.U3 are each independently selected from H and C.sub.1 to C.sub.6 alkyl.

3. The bifunctional molecule according to claim 1, wherein R.sup.3 is a heteroaryl, substituted heteroaryl, aryl, substituted aryl, or a C.sub.1-C.sub.6 alkyl substituted with a heterocyclic group, optionally wherein R.sup.3 is selected from: ##STR00467## wherein the dotted line indicates the position at which each of the respective R.sup.3 groups is joined to the structure shown in formula (I) to (Ic), or wherein when the dotted line is not appended to an atom, the dotted line indicates that each of the respective R.sup.3 groups is joined to the structure via any position on the aromatic or heteroaromatic ring; R.sup.5 is absent or is selected from halo, CF.sub.3, CH.sub.2F, CHF.sub.2, C.sub.1 to C.sub.6 alkyl, CN, OH, OMe, SMe, SOMe, SO.sub.2Me, NH.sub.2, NHMe, NMe.sub.2, CO.sub.2Me, NO.sub.2, CHO and COMe; R.sup.6 is C.sub.1 to C.sub.6 alkyl; and Q is C.sub.1 to C.sub.6 alkylene.

4. The bifunctional molecule according to claim 1, wherein A is CR.sup.2R.sup.2, optionally wherein one of R.sup.2 and R.sup.2 is a hydrogen and the other is C.sub.1 to C.sub.6 alkyl.

5. The bifunctional molecule according to claim 1, wherein B is a phenyl group.

6. The bifunctional molecule according to claim 1, wherein Z is represented as formula (IIaa): ##STR00468## wherein A, R.sup.3, and L are as defined for formula (I); n is 1, 2 or 3; and W is selected from CR.sup.W1R.sup.W2, O, NR.sup.W3 and S; and R.sup.W1, R.sup.W2 and R.sup.W3 are each independently selected from H and C.sub.1 to C.sub.6 alkyl; and wherein when n is 2 or 3, each W is independently selected from CR.sup.W1R.sup.W2, O, NR.sup.W3, and S.

7. The bifunctional molecule according to claim 1, wherein Z is represented as formula (IIa) ##STR00469## wherein R.sup.2, R.sup.2, R.sup.3 and L are as defined in claim 2, n is 1, 2 or 3; and W is selected from CR.sup.W1R.sup.W2, Q, NR.sup.W3 and S; and R.sup.W1, R.sup.W2 and R.sup.W3 are each independently selected from H and C.sub.1 to C.sub.6 alkyl; and wherein when n is 2 or 3, each W is independently selected from CR.sup.W1R.sup.W2, Q, NR.sup.W3, and S.

8. The bifunctional molecule according to claim 1, wherein Z is represented as formula (IIb) ##STR00470## wherein R.sup.2, R.sup.3 and L are as defined in claim 1; m is 3, 4 or 5; and each T is independently selected from CR.sup.T1R.sup.T2, O, NR.sup.T3 and S; and R.sup.T1, R.sup.T2 and R.sup.T3 are each independently selected from H and C.sub.1 to C.sub.6 alkyl.

9. The bifunctional molecule according to any one of the preceding claim 1, wherein the structure of the linker (L) is:
(L.sub.x).sub.q wherein each Lx represents a subunit of L that is independently selected from CR.sup.L1R.sup.L2, O, C?O, S, SO, SO.sub.2, NR.sup.L3, SONR.sup.L4, SONR.sup.L5C?O, CONR.sup.L6, NR.sup.L7CO, C(R.sup.L8)?C(R.sup.L9), C?C, aryl, substituted aryl, heteroaryl, substituted heteroaryl, carbocyclyl, substituted carbocyclyl, heterocyclyl and substituted heterocyclyl groups; wherein R.sup.L1, R.sup.L2, R.sup.L3, R.sup.L4, R.sup.L5, R.sup.L6, R.sup.L7, R.sup.L8 and R.sup.L9 are each independently selected from H, halo, C.sub.1 to C.sub.6 alkyl, C.sub.1 to C.sub.6, haloalkyl, OH, O(C.sub.1 to C.sub.6 alkyl), NH.sub.2, NH(C.sub.1 to C.sub.6 alkyl), NO.sub.2, CN, CONH.sub.2, CONH(C.sub.1 to C.sub.6 alkyl), CON(C.sub.1 to C.sub.6 alkyl).sub.2, SO.sub.2(C.sub.1 to C.sub.6 alkyl), CO.sub.2(C.sub.1 to C.sub.6 alkyl), and CO(C.sub.1 to C.sub.6 alkyl); and q is an integer between 1 and 30.

10. The bifunctional molecule according to claim 1, wherein the target protein binding ligand (TBL) is selected from the group consisting of (i) binders to kinases, (ii) compounds binding to bromodomain-containing proteins, (iii) epigenetic modulator compounds, (iv) binders to transcription factors, (v) binders to GTPases, (vi) binders of phosphatases, (vii) binders of ubiquitin E3 ligases, (viii) immunosuppressive and immunomodulatory compounds, (ix) modulators of nuclear receptors, (x) binders to aggregation-prone proteins, (xi) binders to apoptotic & anti-apoptotic factors, and (xii) binders to polymerases.

11. A pharmaceutical composition comprising the bifunctional molecule according to claim 10, together with a pharmaceutically acceptable carrier, optionally wherein the bifunctional molecule is present in the composition as a pharmaceutically acceptable salt, solvate or derivative.

12. The bifunctional molecule according to claim 10, for use in medicine.

13. The bifunctional molecule for use of claim 12, wherein the use comprises the treatment and/or prevention of any disease or condition which is associated with and/or is caused by an abnormal level of protein activity.

14. The bifunctional molecule of claim 12, for use in the treatment and/or prevention of cancer.

15. A method of selectively degrading and/or increasing proteolysis of a target protein in a cell, the method comprising contacting and/or treating the cell with a bifunctional molecule as defined in claim 1.

16. A method of selectively degrading and/or increasing proteolysis of a target protein in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a bifunctional molecule as defined in claim 1.

17. Use of a moiety Z as defined in any one of formula (I) to (IIa) in a method of targeted protein degradation.

18. Use of a moiety Z as defined in any one of formula (I) to (IIa) in the manufacture of a bifunctional molecule suitable for targeted protein degradation.

19. A compound comprising the Z moiety according to formula (IV): ##STR00471## wherein A, B, R.sup.1, R.sup.3 and R.sup.4 are as defined in claim 1; and G is configured to enable attachment of the Z moiety to another chemical structure via formation of a new covalent bond.

20. A compound comprising the structure:
L-Z wherein Z is as defined in claim 8; and L is a linker.

21. A method of making a bifunctional molecule as defined in claim 1.

22. A method of obtaining bifunctional molecules according to claim 1, comprising: a. providing a bifunctional molecule comprising: (i) a first ligand comprising a structure according to Z as defined in any one of claims 1 to 8; (ii) a second ligand that binds to a target protein; and (iii) a linker that covalently attaches the first and second ligands; b. contacting a cell with the bifunctional molecule; c. detecting degradation of the target protein in the cell; d. detecting degradation of the target protein in the cell in the absence of the bifunctional molecule; and e. comparing the level of degradation of the target protein in the cell contacted with the bifunctional molecule to the level of degradation of the target protein in the absence of the bifunctional molecule; wherein an increased level of degradation of the target protein in the cell contacted with the bifunctional molecule indicates that the bifunctional molecule has facilitated and/or promoted the degradation of the target protein, optionally wherein detecting degradation of the target protein comprises detecting changes in the levels of the target protein in the cell.

23. A compound library comprising a plurality of bifunctional molecules according to claim 1.

Description

DETAILED DESCRIPTION

[0398] The present invention will now be described in detail with reference to the following figures which show:

[0399] FIG. 1 shows plot of correlation between the IC.sub.50 and DC.sub.50 values for a number of bifunctional molecules that are useful in targeted protein degradation.

[0400] FIG. 2 shows log ratio of the GI.sub.50 determined for I-BET726 versus the GI.sub.50 determined for compound A2 plotted for each cell line tested (bars, left y axis). Values>0 indicate cell lines where BET-degradation by A2 shows greater efficacy than the inhibitor I-BET726 due to catalytic activity, whereas values<0 indicate cell lines where BET degrader A2 is less efficacious than BET-inhibition with I-BET726 suggestive of weaker protein degradation.

PART ASYNTHETIC METHODS

[0401] ##STR00101##

##STR00102##

##STR00103##

##STR00104##

##STR00105##

##STR00106##

##STR00107##

##STR00108##

##STR00109##

##STR00110##

##STR00111##

##STR00112##

##STR00113##

##STR00114##

##STR00115##

##STR00116##

##STR00117##

##STR00118## ##STR00119##

##STR00120## ##STR00121##

##STR00122## ##STR00123## ##STR00124##

##STR00125##

##STR00126##

##STR00127##

##STR00128##

##STR00129## ##STR00130##

##STR00131##

##STR00132##

##STR00133##

##STR00134##

[0402] N-AcylationGeneral Procedure 1

##STR00135##

[0403] A solution of phenethylamine (1) (1.0 equiv.) in CH.sub.2Cl.sub.2 (0.2 M) was treated with Et.sub.3N (2.0 equiv.) and acylating agent (II) (e.g. acetic anhydride, or acyl chloride (1.0 equiv.)) at 0? C. After full conversion of the starting material was observed the reaction was quenched by addition of water. The mixture was extracted with CH.sub.2Cl.sub.2 and the combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding N-acylated product (III).

[0404] Bischler-Napieralski CyclisationGeneral Procedure 2

##STR00136##

[0405] A solution of N-Acylphenethylamine (I) (1.0 equiv.) in toluene (0.1 M) was treated with POCl.sub.3 (5.0 equiv.) and heated to 120? C. for 5 hours. The resulting pale yellow solution was cooled to rt and poured onto ice. The biphasic mixture was adjusted to pH?10 with NaOH (4 M in water). The mixture was extracted with EtOAc and the combined organic phases were washed with brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding cyclised imine (II).

[0406] Imine ReductionGeneral Procedure 3

##STR00137##

[0407] A solution of dihydroisoquinoline (1) (1.0 equiv.) in MeOH (0.1 M) (or EtOH (0.1 M) and acetic acid (0.5 equiv.) was treated with sodium borohydride (2.0 equiv.) at 0? C. and stirred for 2 hours. LC-MS showed complete conversion. The reaction was quenched by addition of HCl (1 M) and then adjusted to pH?8 with NaOH and extracted with DCM. The combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure to yield the corresponding tetrahydroisoquinoline.

[0408] DemethylationGeneral Procedure 4

##STR00138##

[0409] A suspension of phenol methyl ether (1) (1.0 equiv.) in 48% aqueous HBr (10 equiv.) was heated to 100? C. for 6 h. The volatiles were evaporated under reduced pressure and the residue was dried until a solid was obtained. Trituration from EtOAc yielded the corresponding phenol (II).

[0410] Boc ProtectionGeneral Procedure 5

[0411] A solution of amine (1.0 equiv.) in THF or DCM (0.1 M) was treated with di-tert-butyl dicarbonate (1.1 equiv.) and saturated aqueous solution of sodium hydrogen carbonate or triethylamine (5.0 equiv.) and the reaction mixture was stirred at rt for 3 h. The reaction mixture was diluted with water and extracted with EtOAc. The combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding Boc-carbamate.

[0412] Phenol AlkylationGeneral Procedure 6

[0413] A solution of phenol (1.0 equiv.) in DMF (0.2 M) was treated with K.sub.2CO.sub.3 (3.0 equiv.) and alkyl halide (2.0 equiv.) and the mixture was stirred for 2 h. The mixture was diluted with EtOAc and washed with water, LiCl (5%) and brine. Purification by flash chromatography yielded the corresponding phenol ether (II).

[0414] Ester HydrolysisGeneral Procedure 7

[0415] A solution of ester (1.0 equiv.) in THF (0.2 M) was treated with lithium hydroxide monohydrate (3.0 equiv.) dissolved in water and the mixture was stirred for 4 h. The mixture was adjusted to pH?3 by addition of 5% KHSO.sub.4 and extracted with EtOAc. The combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure to yield the corresponding carboxylic acid.

[0416] Boc DeprotectionGeneral Procedure 8

[0417] A solution of Boc protected amine (1.0 equiv.) in CH.sub.2Cl.sub.2 (0.05 M) was treated with HCl (4 M in dioxane, 50 equiv.) and the mixture was stirred for 2 h. The volatiles were evaporated under reduced pressure to yield the corresponding amine hydrochloride.

[0418] Amine Cyano AcetylationGeneral Procedure 9

[0419] A suspension of amine (1.0 equiv.) in 1,4-dioxane (0.05 M) was treated with Et.sub.3N or DIPEA (4.0 equiv.) and 3-(3,5-dimethyl-1H-pyrazol-1-yl)-3-oxopropanenitrile (1.1 equiv.) and the mixture was heated to 90? C. for 2 h. The volatiles were evaporated under reduced pressure. Purification by flash chromatography yielded the corresponding cyanoacetamide.

[0420] Knoevenagel CondensationGeneral Procedure 10

[0421] A solution of cyanoacetamide (1.0 equiv.) in THF or EtOH (0.1 M) was treated with aldehyde (2.5 equiv.) and piperidine (0.5 equiv.) and the mixture was heated to reflux for 72 h (if in THF) or at room temperature for 16 h (if in EtOH). The volatiles were evaporated under reduced pressure. Purification by flash chromatography or reverse phase preparative HPLC yielded the corresponding cyanoacrylamide.

[0422] Cbz ProtectionGeneral Procedure 11

[0423] A solution of amine (1.0 equiv.) in THF (0.1 M) was treated with benzyl chloroformate (1.1 equiv.) and saturated aqueous solution of sodium hydrogen carbonate (5.0 equiv.) and the reaction mixture was stirred at rt for 3 h. The reaction mixture was diluted with water and extracted with EtOAc. The combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding Cbz-carbamate.

[0424] Cbz CleavageGeneral Procedure 12

[0425] A solution of Cbz-carbamate (1.0 equiv.) in EtOAc (0.1 M) was treated with Pd/C (10%, 0.05 equiv.) and the suspension was stirred for 2 h under balloon pressure hydrogen atmosphere. The solids were removed by filtration over celite and the filtrate was concentrated under reduced pressure to yield the corresponding amine.

[0426] Amide Coupling with Substituted Acrylic Acid and tBu Ester DeprotectionGeneral Procedure 13

##STR00139##

[0427] A solution of amine (I) (1.0 equiv.) in DMF (0.1 M) was treated with a solution of acrylic acid (II) (1.0 equiv.) HATU (1.1 equiv.) and DIPEA (2.5 equiv.) in DMF (0.1 M) and the reaction mixture was stirred at rt for 15 min. The reaction was quenched with water and extracted with EtOAc. The combined organic layers were washed with LiCl (5%), water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding amide. The resulting amide (1.0 equiv.) in CH.sub.2Cl.sub.2 (0.05 M) was treated with TFA (50 equiv.) and the reaction mixture was stirred at rt for 1 h. The volatiles were evaporated and repeated concentration from CH.sub.2Cl.sub.2 solution followed by prolonged drying yielded the corresponding carboxylic acid (III).

[0428] Doebner-KnoevenagelGeneral Procedure 14

[0429] A solution of aldehyde (1.0 equiv.) and malonic acid (6.0 equiv.) in pyridine (1.5 M) was treated with piperidine (0.1 equiv.) and the reaction mixture was heated to 100? C. for 14 h. The volatiles were evaporated, and the residue was treated with HCl (1 M). The precipitate was collected by filtration to yield the corresponding acrylic acid.

[0430] Fluoroacrylate Ester SynthesisGeneral Procedure 15

##STR00140##

[0431] A solution of aldehyde (II) (1.0 equiv.) and 2-fluoro-3-oxo-3-phenylpropionate (I) (1.2 equiv.) in MeCN (0.3 M) was treated with Cs.sub.2CO.sub.3 (2.0 equiv.) and the reaction mixture was heated to 40? C. for 14 h. The reaction mixture was cooled to rt, diluted with EtOAc and filtered over celite. Purification by flash chromatography yielded the corresponding fluoroacrylate ester. The ester was then treated with sodium hydroxide (2 M, 2 equiv.) in THF and left to react until complete deprotection. The mixture was treated with KHSO.sub.4 (10%) to pH=3. The precipitate was filtered and dried to obtain fluoroacrylate (III).

[0432] Trifluoromethyl Acrylic Acid SynthesisGeneral Procedure 16

##STR00141##

[0433] A solution of aldehyde (II) (1.5 equiv.) and 3,3,3-trifluoropropionic acid (I) (1.0 equiv.) in THF (0.2 M) was cooled to 0? C. and treated with TiCl.sub.4 (1.0 M in CH.sub.2Cl.sub.2, 2.0 equiv.) and the mixture was stirred for 30 min. The resulting solution was treated with Et.sub.3N (4.0 equiv.), warmed to rt and stirred for 72 h. The reaction was quenched by addition of water and extracted with CH.sub.2Cl.sub.2. The combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding trifluoromethyl acrylic acid (III).

[0434] Malonamic Amide Knoevenagel CondensationGeneral Procedure 17

##STR00142##

[0435] A solution of N,N-dimethylmalonamic acid tert-butyl ester (1) (1.0 equiv.) in THF (0.2 M) was treated with aldehyde (II) (2.5 equiv.) and piperidine (0.5 equiv.) and the mixture was heated to 66? C. for 72 h. The volatiles were evaporated under reduced pressure. Purification by flash chromatography yielded the corresponding amido-acrylamide tert-butyl ester. The ester was then treated with a 50% solution of TFA in DCM and left to react until complete deprotection of the tert-butyl ester was observed by HPLC. Volatiles were removed under reduced pressure to obtain malonamic amide (III).

[0436] Sulfone Knoevenagel CondensationGeneral Procedure 18

##STR00143##

[0437] A solution of tert-butyl methylsulfonyl acetate (1) (1.0 equiv.) in THF (0.2 M) was treated with aldehyde (II) (2.5 equiv.) and piperidine (0.5 equiv.) and the mixture was heated to 66? C. for 72 h. The volatiles were evaporated under reduced pressure. Purification by flash chromatography yielded the corresponding sulfonacrylate ester. The ester was then treated with a 50% solution of TFA in DCM and left to react until complete deprotection of the tertbutyl ester was observed by HPLC. Volatiles were removed under reduced pressure and the acid was crystallised from ethyl acetate to obtain sulfonacrylate (III).

[0438] TBS ProtectionGeneral Procedure 19

[0439] A solution of alcohol (1.0 equiv.) in CH.sub.2Cl.sub.2 was treated with TBSCI (1.3 equiv.) and imidazole (1.0 equiv.). The colourless suspension was stirred at rt for 4 h. The reaction mixture was quenched by addition of NH.sub.4Cl (sat. aq.) and extracted with CH.sub.2Cl.sub.2. The combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding silylether.

[0440] Sulfinimide FormationGeneral Procedure 20

[0441] A solution of aldehyde or ketone (1.0 equiv.) in THF was treated with titanium (IV) ethoxide (2.0 equiv.) followed by 2-methylpropane-2-sulfinamide (1.3 equiv.) and the mixture was heated to 66? C. for 16 h. The reaction was quenched by addition of NH.sub.4Cl (sat. aq.), diluted with EtOAc and the precipitated solids were removed by filtration. The biphasic mixture was extracted with EtOAc, and the combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding sulfinimide.

[0442] Sulfinimine ReductionGeneral Procedure 21

[0443] A solution of sulfinimide (1.0 equiv.) in THF (0.1 M) was cooled to 0? C. and treated with NaBH.sub.4 (2.0 equiv.) and the mixture was stirred for 2 h and then warmed to rt. The reaction was quenched by addition of NH.sub.4Cl (sat. aq.) and extracted with EtOAc. The combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding sulfinamide.

[0444] Grignard Addition to SulfinimineGeneral Procedure 22

[0445] A solution of sulfinimide (1.0 equiv.) in CH.sub.2Cl.sub.2 was cooled to ?78? C. and treated with organomagnesium halide (II) (2.0 equiv.) and the mixture was stirred for 2 h and then warmed to rt. The reaction was quenched by addition of NH.sub.4Cl (sat. aq.) and extracted with EtOAc. The combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding sulfinamide.

[0446] Sulfinamide AlkylationGeneral Procedure 23

[0447] A solution of sulfinamide (1.0 equiv.) in THF was cooled to 0? C. and treated with LiHMDS (2.0 equiv.) and the mixture was stirred for 15 min. The resulting solution was treated with organohalide, and warmed to rt. The reaction was quenched by addition of water and extracted with EtOAc. The combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding N-alkylated sulfinamide.

[0448] Silylether CleavageGeneral Procedure 24

[0449] A solution of silyl ether (1.0 equiv.) in CH.sub.2Cl.sub.2 was treated with TBAF (1 M in THF, 1.2 equiv.) and the mixture was stirred for 12 h. The reaction was quenched by addition of NH.sub.4Cl (sat. aq.) and extracted with CH.sub.2Cl.sub.2. The combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding alcohol.

[0450] Sulfinamide DeprotectionGeneral Procedure 25

[0451] To a solution of sulfinamide (1.0 equiv.) in diethyl ether (0.2 M) was added HCl (4 M in dioxane, 3.0 equiv.). The reaction was monitored by TLC. Once no starting material was observed, the suspension was filtered, washed with diethyl ether and the isolated amine hydrochloride salt was used without further purification.

[0452] Reductive AminationGeneral Procedure 26

[0453] A solution of amine (1.0 equiv.) in MeOH (or THF or DCM) (0.1 M) was treated with ketone or aldehyde (2.0 equiv.) and sodium cyanoborohydride (or sodium triacetoxyborohydride, or polymer-bound cyanoborohydride) (4.0 equiv.) and the mixture was stirred for 12 h. The reaction was quenched by addition of water and extracted with EtOAc. The combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding amine.

[0454] Suzuki CouplingGeneral Procedure 27

[0455] A solution of aryl halide (1.0 equiv.), boronic acid or ester (1.5 equiv.), K.sub.2CO.sub.3 (2.5 equiv.) and Pd(dppf)Cl.sub.2 (0.1 equiv.) in 1,4-dioxane/water was deoxygenated by sparging with N.sub.2 and consequently heated to 100? C. for 1 h. The reaction was cooled to rt, diluted with EtOAc and the solids were removed by filtration. The residue was concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding coupling product.

[0456] Alkene HydrogenationGeneral Procedure 28

[0457] A solution of alkene (1.0 equiv.) in MeOH was treated with Pd/C (10%, 0.05 equiv.) and the suspension was stirred for 2 h under hydrogen atmosphere. The solids were removed by filtration over celite and the filtrate was concentrated under reduced pressure to yield the corresponding hydrogenation product.

[0458] Ester Reduction to AldehydeGeneral Procedure 29

[0459] A solution of ester (1.0 equiv.) in CH.sub.2Cl.sub.2 (0.1 M) was cooled to ?78? C. and treated with DIBAL-H (1 M in heptane, 1.2 equiv.) and the resulting solution was stirred for 2 h. The reaction was quenched by addition of MeOH at ?78? C., and warmed to rt. The reaction was diluted with CH.sub.2Cl.sub.2 and treated with Rochelle's salt (10% aq.) and stirred until a clear biphasic mixture was obtained. The mixture was extracted with CH.sub.2Cl.sub.2, the combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding aldehyde.

[0460] Amine AlkylationGeneral Procedure 30

[0461] To a stirred solution of amine (1 equiv.), DIPEA (3.0 equiv.) and potassium iodide (0.3 equiv.) in DMF was added alkyl-Br (1 equiv.) at RT. The reaction mixture was stirred at RT for 5 h then diluted in water and extracted with EtOAc. The combined organic layers were dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude product was purified by flash chromatography to give the desired product.

[0462] Amide CouplingGeneral Procedure 31

[0463] To a stirred solution of carboxylic acid (1.0 equiv.) in DMF was added DIPEA (2.5 equiv.) and HATU (1.5 equiv.). The reaction mixture was stirred for 5 min, then relevant amine (1.5 equiv.) was added and the reaction mixture was stirred for 12 h at RT. The reaction was quenched with ice cold water and extracted with EtOAc. The combined organic layers were concentrated in vacuo to afford the crude product. Where stated, the crude product was purified by flash chromatography/reverse phase preparative HPLC to give the desired compound.

[0464] Reduction of Weinreb Amide to Aldehyde Using DIBALHGeneral Procedure 32

[0465] To a stirred solution of Weinreb amide (1.0 equiv.) in THF at 0? C. was added DIBALH (3.0 equiv.) dropwise. The reaction was stirred for 4 h at RT then was quenched with HCl (1.5 N) and water, then filtered through celite. The filtrate was extracted with EtOAc, and the combined organic layers were dried (Na.sub.2SO.sub.4), filtered and concentrated in vacuo to give the desired compound

[0466] Tandem Boc/tBu DeprotectionGeneral Procedure 33:

[0467] A solution of N-Boc,CO.sub.2t-Bu-amino acid (1) (1.0 equiv.) in DCM (0.1 M) was treated with HCl (4 M in dioxane, 50 equiv.) and the mixture was stirred at rt for 48 h. The volatiles were evaporated under reduced pressure to yield the corresponding amino acid.

[0468] Reductive Amination with Ammonium Acetate/Acylation: General Procedure 34:

[0469] A solution of ketone (1.0 equiv.) in methanol (0.15 M) at rt was treated with ammonium acetate (10.0 equiv.) followed by sodium cyanoborohydride (1.50 equiv.) and stirred for 12 h. The reaction was quenched with water and the mixture was extracted with DCM. The combined organic extracts were washed with brine, dried over MgSO.sub.4 and concentrated under reduced pressure. The crude residue was redissolved in DCM (0.15 M), cooled to 0? C., treated with triethylamine (2.00 equiv.) and acyl chloride (1.50 equiv.) and stirred for 2 h. The reaction was quenched with water and extracted with DCM. The combined organic extracts were washed with brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the N-acylated product.

[0470] FeCl.sub.3-Mediated Bischler-Napieralski Cyclisation: General Procedure 35:

[0471] To a stirred solution of N-acylphenethylamine (1.00 equiv.) in DCM (0.3 M) at 0? C. was added drop wise oxalyl chloride (12.0 equiv.). The resulting reaction mixture was warmed to rt and stirred for 2 h. The reaction mixture was then cooled to ?78? C., and iron(III) chloride (6.00 equiv.) was added portion wise. The mixture was warmed to rt slowly and stirred for 16 h. The reaction was quenched with HCl (1 M) and extracted with DCM. The combined extracts were concentrated under reduced pressure. The residue was redissolved in H.sub.2SO.sub.4:MeOH (1:10, 0.1 M) and stirred at 80? C. for 16 h. The mixture was concentrated under reduced pressure and redissolved in water. The pH was adjusted to ?7 with 25% aq. ammonia and the mixture was extracted with DCM. The combined extracts were washed with brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the cyclised imine.

[0472] Preparative HPLC conditions: a Xbridge C18 (19?150 mm) 5 ?m silica_column was used. When not specified otherwise, a 5-95% gradient of acetonitrile in 10 mM ammonium acetate was used, with a flow rate of 15 ml/min.

PREPARATIVE EXAMPLES

Example 1: N-(3-methoxyphenethyl)acetamide

[0473] ##STR00144##

[0474] Prepared following general procedure 1. Obtained 1.46 g, 79.4% yield.

[0475] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.23 (t, J=7.8 Hz, 1H), 6.82-6.76 (m, 2H), 6.74 (t, J=2.0 Hz, 1H), 5.48 (s, 1H), 3.80 (s, 3H), 3.52 (td, J=6.9, 5.8 Hz, 2H), 2.79 (t, J=6.9 Hz, 2H), 1.94 (s, 3H).

Example 2: N-(3-methoxyphenethyl)butyramide

[0476] ##STR00145##

[0477] Prepared following general procedure 1. Obtained 870 mg, 65.2% yield.

[0478] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.23 (t, J=7.9 Hz, 1H), 6.86-6.72 (m, 3H), 5.40 (s, 1H), 3.80 (s, 3H), 3.53 (td, J=6.9, 5.8 Hz, 2H), 2.79 (t, J=6.9 Hz, 2H), 2.10 (dd, J=8.2, 6.8 Hz, 2H), 1.62 (dt, J=14.6, 7.3 Hz, 2H), 0.92 (t, J=7.4 Hz, 3H).

Example 3: 6-methoxy-1-methyl-3,4-dihydroisoquinoline

[0479] ##STR00146##

[0480] Prepared following general procedure 2. Obtained 446 mg, 76.6% yield.

[0481] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.47 (d, J=8.6 Hz, 1H), 6.81 (dd, J=8.6, 2.7 Hz, 1H), 6.72 (d, J=2.4 Hz, 1H), 3.85 (s, 3H), 3.67 (tq, J=7.5, 1.5 Hz, 2H), 2.74 (dd, J=8.5, 6.5 Hz, 2H), 2.43 (s, 3H).

Example 4: 6-methoxy-1-propyl-3,4-dihydroisoquinoline

[0482] ##STR00147##

[0483] Prepared following general procedure 2. Obtained 553 mg, 69.2% yield.

[0484] .sup.1H NMR (400 MHz, CDCl3) ? 7.51 (d, J=8.6 Hz, 1H), 6.83 (dd, J=8.6, 2.7 Hz, 1H), 6.75 (d, J=2.6 Hz, 1H), 3.86 (s, 3H), 3.70 (t, J=7.5 Hz, 2H), 2.89-2.67 (m, 4H), 1.72 (h, J=7.4 Hz, 2H), 1.01 (t, J=7.4 Hz, 3H).

Example 5: 6-methoxy-1-methyl-1,2,3,4-tetrahydroisoquinoline

[0485] ##STR00148##

[0486] Prepared following general procedure 3. Obtained 417 mg, 92.8% yield

[0487] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.06 (dd, J=8.6, 0.8 Hz, 1H), 6.74 (dd, J=8.6, 2.8 Hz, 1H), 6.62 (d, J=2.8 Hz, 1H), 4.10 (qd, J=6.4, 0.8 Hz, 1H), 3.78 (s, 3H), 3.28 (dt, J=12.4, 5.1 Hz, 1H), 3.04 (ddd, J=12.5, 8.9, 4.7 Hz, 1H), 2.96-2.84 (m, 1H), 2.74 (dt, J=16.4, 4.7 Hz, 1H), 2.40 (s, 1H), 1.47 (d, J=6.7 Hz, 3H).

Example 6: 6-methoxy-1-propyl-1,2,3,4-tetrahydroisoquinoline

[0488] ##STR00149##

[0489] Prepared following general procedure 3. Obtained 452 mg, 80.9% yield.

[0490] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.05 (d, J=8.6 Hz, 1H), 6.72 (dd, J=8.5, 2.8 Hz, 1H), 6.61 (d, J=2.7 Hz, 1H), 3.94 (dd, J=9.2, 3.8 Hz, 1H), 3.82-3.79 (m, 1H), 3.78 (s, 3H), 3.24 (dt, J=12.4, 5.4 Hz, 1H), 2.98 (ddd, J=12.5, 7.8, 5.0 Hz, 1H), 2.87-2.78 (m, 1H), 2.72 (dt, J=16.3, 5.2 Hz, 1H), 1.87-1.62 (m, 2H), 1.58-1.36 (m, 2H), 0.97 (t, J=7.3 Hz, 3H).

Example 7: 1-methyl-1,2,3,4-tetrahydroisoquinolin-6-ol hydrobromide

[0491] ##STR00150##

[0492] Prepared following general procedure 4. Obtained 492 mg, 85.7% yield.

[0493] .sup.1H NMR (400 MHz, DMSO) ? 9.48 (s, 1H), 9.05 (s, 1H), 8.70 (s, 1H), 7.09 (d, J=8.5 Hz, 1H), 6.68 (dd, J=8.5, 2.6 Hz, 1H), 6.58 (d, J=2.5 Hz, 1H), 4.44 (s, 1H), 3.41 (s, 2H), 2.92 (qt, J=11.3, 6.2 Hz, 2H), 1.52 (d, J=6.8 Hz, 3H).

Example 8: 1-propyl-1,2,3,4-tetrahydroisoquinolin-6-ol hydrobromide

[0494] ##STR00151##

[0495] Prepared following general procedure 4. Obtained 485 mg, 80.9% yield.

[0496] .sup.1H NMR (400 MHz, DMSO) ? 9.49 (s, 1H), 9.05 (s, 1H), 8.55 (s, 1H), 7.07 (d, J=8.6 Hz, 1H), 6.67 (dd, J=8.5, 2.6 Hz, 1H), 6.A (d, J=2.6 Hz, 1H), 4.35 (s, 1H), 3.43-3.36 (m, 1H), 3.25 (dd, J=12.7, 6.5 Hz, 1H), 2.91 (dtt, J=17.6, 12.6, 6.6 Hz, 2H), 1.95-1.70 (m, 2H), 1.42 (ddt, J=13.5, 9.4, 6.7 Hz, 2H), 0.94 (t, J=7.3 Hz, 3H).

Example 9: tert-butyl 6-hydroxy-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0497] ##STR00152##

[0498] Prepared following general procedure 5. Obtained 485 mg, 80.9% yield.

[0499] .sup.1H NMR (400 MHz, DMSO) ? 9.20 (s, 1H), 6.97 (d, J=8.4 Hz, 1H), 6.58 (dd, J=8.3, 2.6 Hz, 1H), 6.50 (d, J=2.5 Hz, 1H), 4.94 (s, 1H), 3.98-3.77 (m, 1H), 3.23-3.00 (m, 1H), 2.73-2.56 (m, 2H), 1.42 (s, 9H), 1.30 (d, J=6.7 Hz, 3H).

Example 10: tert-butyl 6-hydroxy-1-propyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0500] ##STR00153##

[0501] Prepared following general procedure 5. Obtained 216 mg, 80.7% yield.

[0502] .sup.1H NMR (400 MHz, DMSO) ? 9.19 (s, 1H), 6.92 (d, J=8.4 Hz, 1H), 6.56 (dd, J=8.3, 2.6 Hz, 1H), 6.49 (d, J=2.5 Hz, 1H), 4.87 (d, J=27.0 Hz, 1H), 3.95-3.75 (m, 1H), 3.24-3.01 (m, 2H), 2.74-2.57 (m, 1H), 1.67 (s, 1H), 1.60-1.47 (m, 1H), 1.40 (s, 9H), 1.34-1.24 (m, 2H), 0.89 (t, J=9.3 Hz, 3H).

Example 11: tert-butyl 6-(2-ethoxy-2-oxoethoxy)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0503] ##STR00154##

[0504] Prepared following general procedure 6. Obtained 229 mg, 79.9% yield.

[0505] .sup.1H NMR (400 MHz, CDCl3) ? 7.03 (d, J=8.6 Hz, 1H), 6.76 (dd, J=8.4, 2.6 Hz, 1H), 6.64 (d, J=2.7 Hz, 1H), 5.25-5.01 (m, 1H), 4.59 (s, 2H), 4.27 (q, J=7.1 Hz, 2H), 4.20-3.96 (m, 1H), 3.30-3.06 (m, 1H), 2.92-2.80 (m, 1H), 2.68 (dt, J=16.1, 3.6 Hz, 1H), 1.48 (s, 9H), 1.40 (d, J=6.7 Hz, 3H), 1.30 (t, J=7.1 Hz, 3H).

Example 12: tert-butyl 6-(2-ethoxy-2-oxoethoxy)-1-propyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0506] ##STR00155##

[0507] Prepared following general procedure 6. Obtained 244 mg, 87.2% yield.

[0508] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.01 (d, J=8.4 Hz, 1H), 6.74 (d, J=8.4 Hz, 1H), 6.64 (d, J=2.7 Hz, 1H), 5.15-4.92 (m, 1H), 4.58 (s, 2H), 4.32-4.18 (m, 2H), 3.97-3.83 (m, 1H), 3.20-3.07 (m, 1H), 2.96-2.79 (m, 1H), 2.71-2.62 (m, 1H), 1.84-1.70 (m, 1H), 1.65-1.59 (m, 1H), 1.45 (s, 9H), 1.45-1.35 (m, 2H), 1.32-1.28 (m, 3H), 0.95 (t, J=7.4 Hz, 3H).

Example 13: benzyl 6-(2-(tert-butoxy)-2-oxoethoxy)-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0509] ##STR00156##

[0510] Prepared following general procedure 6. Obtained 330 mg, 84.0% yield. .sup.1H NMR (400 MHz, DMSO) ? 7.41-7.27 (m, 5H), 7.10 (d, J=8.4 Hz, 1H), 6.77-6.67 (m, 2H), 5.12 (s, 2H), 4.60 (s, 2H), 4.56-4.43 (m, 2H), 3.60 (s, 2H), 2.76 (t, J=6.0 Hz, 2H), 1.42 (s, 9H).

Example 14: 2-((2-(tert-butoxycarbonyl)-1-methyl-1,2,3,4-tetrahydroisoquinolin-6-yl)oxy)acetic acid

[0511] ##STR00157##

[0512] Prepared following general procedure 7. Obtained 51.3 mg, 97.0% yield.

[0513] .sup.1H NMR (400 MHz, DMSO) ? 12.95 (s, 1H), 7.11 (d, J=8.6 Hz, 1H), 6.73 (dd, J=8.5, 2.8 Hz, 1H), 6.66 (d, J=2.7 Hz, 1H), 5.00 (s, 1H), 4.61 (s, 2H), 3.92 (s, 2H), 3.12 (s, 2H), 1.42 (s, 9H), 1.32 (d, J=6.6 Hz, 3H).

Example 15: 2-((2-(tert-butoxycarbonyl)-1-propyl-1,2,3,4-tetrahydroisoquinolin-6-yl)oxy)acetic acid

[0514] ##STR00158##

[0515] Prepared following general procedure 7. Obtained 48.8 mg, 86% yield.

[0516] .sup.1H NMR (400 MHz, DMSO) ? 12.95 (s, 1H), 7.06 (d, J=8.5 Hz, 1H), 6.72 (dd, J=8.5, 2.8 Hz, 1H), 6.66 (d, J=2.7 Hz, 1H), 4.99-4.86 (m, 1H), 4.62 (s, 2H), 3.99-3.78 (m, 1H), 3.25-3.05 (m, 1H), 2.75-2.64 (m, 2H), 1.75-1.51 (m, 2H), 1.41 (s, 9H), 1.34-1.23 (m, 2H), 0.97-0.85 (m, 3H).

Example 16: benzyl 6-hydroxy-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0517] ##STR00159##

[0518] Prepared following general procedure 11. Obtained 558 mg, 74% yield. .sup.1H NMR (400 MHz, DMSO) ? 9.22 (s, 1H), 7.40-7.27 (m, 5H), 6.96 (d, J=8.3 Hz, 1H), 6.58 (dd, J=8.2, 2.5 Hz, 1H), 6.55 (d, J=2.5 Hz, 1H), 5.11 (s, 2H), 4.45 (d, J=16.0 Hz, 2H), 3.57 (s, 2H), 2.70 (t, J=6.0 Hz, 2H).

Example 17: tert-butyl 2-((1,2,3,4-tetrahydroisoquinolin-6-yl)oxy)acetate

[0519] ##STR00160##

[0520] Prepared following general procedure 12. Obtained 233 mg, 92.5% yield. .sup.1H NMR (400 MHz, DMSO) ? 6.90 (d, J=8.4 Hz, 1H), 6.63 (dd, J=8.4, 2.8 Hz, 1H), 6.56 (d, J=2.7 Hz, 1H), 4.56 (s, 2H), 3.74 (s, 2H), 2.89 (t, J=5.9 Hz, 2H), 2.62 (t, J=5.9 Hz, 2H), 1.42 (s, 9H).

Example 18: (E)-2-((2-(2-cyano-3-(thiazol-2-yl)acryloyl)-1,2,3,4-tetrahydroisoquinolin-6-yl)oxy)acetic acid

[0521] ##STR00161##

[0522] Prepared following general procedure 13. Obtained 2.2 mg, 95.7% yield. m/z=370.0 (M+H).sup.+.

Example 19: (E)-3-(thiazol-2-yl)acrylic acid

[0523] ##STR00162##

[0524] Prepared following general procedure 14. Obtained 350 mg, 25.5% yield.

[0525] .sup.1H NMR (400 MHz, DMSO) ? 12.79 (s, 1H), 8.00 (d, J=3.2 Hz, 1H), 7.93 (dd, J=3.2, 0.6 Hz, 1H), 7.68 (dd, J=15.8, 0.6 Hz, 1H), 6.66 (d, J=15.8 Hz, 1H).

Example 20: (Z)-3-(6-bromopyridin-2-yl)-2-fluoroacrylic acid

[0526] ##STR00163##

[0527] Prepared following general procedure 15. Obtained 190 mg, 77% yield. m/z=246.0 (M+H).sup.+. .sup.1H NMR (400 MHz, DMSO) ? 14.07 (s, 1H), 7.90-7.80 (m, 2H), 7.66 (dd, J=7.0, 1.7 Hz, 1H), 6.93 (d, J.sub.H-F=34.1 Hz, 1H).

Example 20a: (Z)-2-fluoro-3-(thiazol-2-yl)acrylic acid

[0528] ##STR00164##

[0529] Prepared following general procedure 15. Obtained 88 mg, 56% yield. m/z=174.0 (M+H).sup.+.

Example 21: (Z)-3-(6-bromopyridin-2-yl)-2-(trifluoromethyl)acrylic acid

[0530] ##STR00165##

[0531] Prepared following general procedure 16. Obtained 64.8 mg, 27.5% yield.

[0532] .sup.1H NMR (400 MHz, DMSO) ? 13.82 (s, 1H), 7.86 (t, J=7.8 Hz, 1H), 7.73-7.65 (m, 2H), 7.46 (s, 1H).

Example 22: (E)-2-(dimethylcarbamoyl)-3-(thiazol-2-yl)acrylic acid

[0533] ##STR00166##

[0534] Prepared following general procedure 17. Obtained 14.8 mg, 24.5% yield. m/z=227.2 (M+H).sup.+

Example 23: (Z)-2-(methylsulfonyl)-3-(thiazol-2-yl)acrylic acid

[0535] ##STR00167##

[0536] Prepared following general procedure 18. Obtained 88 mg, 42% yield.

[0537] .sup.1H NMR (400 MHz, DMSO) ? 13.41 (s, 1H), 8.16 (dd, J=3.1, 0.6 Hz, 1H), 8.14 (d, J=3.1 Hz, 1H), 7.84 (d, J=0.7 Hz, 1H), 3.31 (s, 3H).

Example 24: 6-((tert-butyldimethylsilyl)oxy)-3,4-dihydronaphthalen-1(2H)-one

[0538] ##STR00168##

[0539] Prepared following general procedure 19. Obtained 1.35 g, 79.2% yield. m/z=277.2 (M+H).sup.+.

Additional Examples

[0540]

TABLE-US-00001 Example m/z number Structure Preparation (M + H).sup.+ 25 [00169] embedded image Prepared following general procedure 19 263.2 25a [00170] Prepared following general procedure 19 279.2

Example 26: (E)-N-(6-((tert-butyldimethylsilyl)oxy)-3,4-dihydronaphthalen-1(2H)-ylidene)-2-methylpropane-2-sulfinamide

[0541] ##STR00171##

[0542] Prepared following general procedure 20. Obtained 1.12 g, 60.4% yield.

[0543] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 8.10 (d, J=8.7 Hz, 1H), 6.71 (dd, J=8.8, 2.5 Hz, 1H), 6.61 (d, J=2.3 Hz, 1H), 3.23 (ddd, J=17.5, 9.1, 4.8 Hz, 1H), 3.00 (ddd, J=17.5, 7.4, 4.5 Hz, 1H), 2.88-2.72 (m, 2H), 2.08-1.88 (m, 2H), 1.31 (s, 9H), 0.98 (s, 9H), 0.23 (s, 6H).

Additional Examples

[0544]

TABLE-US-00002 Example m/z number Structure Preparation (M + H).sup.+ 26a [00172] embedded image Prepared following general procedure 20 366.2 26b [00173] Prepared following general procedure 20 382.2 26c [00174] Prepared following general procedure 20 330.0

Example 27: N-(6-((tert-butyldimethylsilyl)oxy)-1,2,3,4-tetrahydronaphthalen-1-yl)-2-methylpropane-2-sulfinamide

[0545] ##STR00175##

[0546] Prepared following general procedure 21. Obtained 1.02 g, 90.6% yield. m/z=382.3 (M+H).sup.+

Additional Examples

[0547]

TABLE-US-00003 Example m/z number Structure Preparation (M + H).sup.+ 27a [00176] embedded image Prepared following general procedure 21 368.2 27b [00177] Prepared following general procedure 21 384.2 27c [00178] Prepared following general procedure 21 332.1

Example 28: N-(1-(4-((tert-butyldimethylsilyl)oxy)phenyl)heptyl)-2-methylpropane-2-sulfinamide

[0548] ##STR00179##

[0549] Prepared following general procedure 22. Obtained 663 mg, 86.7% yield.

[0550] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.18-7.09 (m, 2H), 6.84-6.74 (m, 2H), 4.28 (ddd, J=8.5, 6.2, 2.6 Hz, 1H), 3.32 (d, J=2.8 Hz, 1H), 2.05-1.69 (m, 2H), 1.28-1.18 (m, 8H), 1.16 (s, 9H), 0.98 (s, 9H), 0.88-0.78 (m, 3H), 0.19 (s, 6H).

Example 29: N-benzyl-N-(1-(4-((tert-butyldimethylsilyl)oxy)phenyl)butyl)-2-methylpropane-2-sulfinamide

[0551] ##STR00180##

[0552] Prepared following general procedure 23. Obtained 594 mg, 80.2% yield.

[0553] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.42-7.27 (m, 5H), 7.13-7.05 (m, 2H), 6.81-6.72 (m, 2H), 4.53 (d, J=16.4 Hz, 1H), 3.96-3.87 (m, 2H), 2.29-2.14 (m, 1H), 2.15-2.02 (m, 1H), 1.22-1.01 (m, 2H), 0.98 (s, 9H), 0.96 (s, 9H), 0.83 (t, J=7.3 Hz, 3H), 0.18 (s, 6H).

Additional Examples

[0554]

TABLE-US-00004 Example m/z number Structure Preparation (M + H).sup.+ 29a [00181] embedded image Prepared following general procedure 23 398.3 29b [00182] Prepared following general procedure 23 412.3 29c [00183] Prepared following general procedure 23 347.3

Example 30: N-ethyl-N-(1-(4-hydroxyphenyl)butyl)-2-methyl propane-2-sulfinamide

[0555] ##STR00184##

[0556] Prepared following general procedure 24. Obtained 213 mg, 79.5% yield.

[0557] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.19 (d, J=8.5 Hz, 2H), 6.77 (d, J=8.4 Hz, 2H), 5.98 (s, 1H), 4.21 (dd, J=9.6, 5.6 Hz, 1H), 3.27 (dq, J=14.8, 7.4 Hz, 1H), 2.61 (dq, J=14.0, 7.0 Hz, 1H), 2.15-1.95 (m, 2H), 1.35-1.25 (m, 2H), 1.19 (t, J=7.2 Hz, 3H), 1.07 (d, J=0.9 Hz, 9H), 0.92 (t, J=7.4 Hz, 3H).

Additional Examples

[0558]

TABLE-US-00005 Example m/z number Structure Preparation (M + H).sup.+ 30a [00185] embedded image Prepared following general procedure 24 284.2 30b [00186] Prepared following general procedure 24 298.2

Example 31: 1-(4-bromophenyl)-N-methylbutan-1-amine

[0559] ##STR00187##

[0560] Prepared following general procedure 25. Obtained 112 mg, quantitative yield. m/z=211.0 corresponding to the 1-(4-bromophenyl)butan-1-ylium cation.

Example 32

[0561] ##STR00188##

[0562] To a stirred solution of 1-(4-bromophenyl)butan-1-one (50 g, 220 mmol) in methyl amine (2M in THF, 330 ml, 660 mmol) was added Titanium ethoxide (60.5 ml, 286 mmol) at RT under nitrogen atmosphere. The colour of the reaction mixture was turned from colorless to turbid. The reaction mixture was stirred for 16 h at RT. Thereafter, reaction mixture was turned to pale yellow colour when NaBH.sub.4 (8.33 g, 220 mmol) was added to it portionwise manner at 0? C. The resultant reaction mixture was stirred for 4 h at RT. The progress of the reaction was monitored by UPLC. The reaction mixture was diluted with excess of MTBE and quenched with saturated sodium bicarbonate solution. The solid formed of TiO.sub.2 was filtered and the filtrate was concentrated to afford crude 1-(4-bromophenyl)-N-methylbutan-1-amine (52 g, 204 mmol, 93% yield) as pale yellow liquid. This crude used as such without further purification. To a stirred solution of 1-(4-bromophenyl)-N-methylbutan-1-amine (52 g, 215 mmol) in DCM (250 ml) was added N-ethyl-N-isopropylpropan-2-amine (74.8 ml, 429 mmol) at RT. Then after 10 min, di-tert-butyl dicarbonate (74.0 ml, 322 mmol) was added slowly dropwise at RT to the reaction mixture, the reaction mixture turned from colorless to white turbid solution and the resultant reaction mixtured was stirred for 16 h at RT under nitrogen atmosphere. The progress of the reaction was monitored by UPLC. The reaction mixture was concentrated under reduced pressure and the obtained residue was diluted with aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic layer was washed with aqueous sodium bicarbonate solution, brine solution, dried over sodium sulfate and concentrated under reduced pressure. The obtained crude was purified by silica gel column chromatography (100-200 mesh, 0-4% ethyl acetate in n-hexane) to afford tert-butyl (1-(4-bromophenyl)butyl)(methyl)carbamate (57 g, 165 mmol, 77% yield) as colorless oil.

[0563] .sup.1H-NMR (400 MHz, CDCl3): ? 7.46 (d, J=8.40 Hz, 2H), 7.18 (d, J=8.00 Hz, 2H), 5.32 (m, 1H), 2.56 (s, 3H), 1.84 (d, J=7.20 Hz, 2H), 1.53 (s, 9H), 1.37 (q, J=7.60 Hz, 2H), 1.01 (t, J=7.60 Hz, 3H).

Example 33: ethyl 2-(4-(1-(isopropylamino)butyl)phenoxy)acetate

[0564] ##STR00189##

[0565] Prepared following general procedure 26. Obtained 115 mg, 75.2% yield.

[0566] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.42 (d, J=8.2 Hz, 2H), 6.93 (d, J=8.7 Hz, 2H), 4.62 (s, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.95 (d, J=10.3 Hz, 1H), 2.87-2.75 (m, 1H), 2.07 (d, J=51.2 Hz, 2H), 1.31 (t, J=7.1 Hz, 6H), 1.24 (d, J=6.3 Hz, 3H), 1.20-1.00 (m, 2H), 0.85 (t, J=7.3 Hz, 3H).

Example 34: 2-(4-(1-(2-cyano-N-methylacetamido)butyl)phenoxy)acetic acid

[0567] ##STR00190##

[0568] Prepared following general procedure 9. Obtained 230 mg, 81.1% yield. m/z=609.7 (2M+H).sup.+

Additional Example

[0569] ##STR00191##

Example 36: (E)-2-(4-(1-(2-cyano-N-isopropyl-3-(thiazol-2-yl)acrylamido)butyl)phenoxy)acetic acid

[0570] ##STR00192##

[0571] Prepared following general procedure 10. Obtained 34.4 mg, 81.0% yield. m/z=428.4 (M+H).sup.+.

Additional Example

[0572] ##STR00193##

Example 37: ethyl (E)-3-(4-(1-((tert-butoxycarbonyl)(methyl)amino)butyl)phenyl)acrylate

[0573] ##STR00194##

[0574] Prepared following general procedure 27. Obtained 115 mg, 75.2% yield.

[0575] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.65 (d, J=16.0 Hz, 1H), 7.51-7.43 (m, 2H), 7.29 (dd, J=9.7, 4.9 Hz, 2H), 6.41 (d, J=16.0 Hz, 1H), 5.29 (d, J=56.7 Hz, 1H), 4.25 (q, J=7.1 Hz, 2H), 2.55 (s, 3H), 1.84 (dt, J=13.0, 6.3 Hz, 2H), 1.47 (s, 9H), 1.43-1.33 (m, 2H), 1.32 (t, J=7.1 Hz, 3H), 0.98 (t, J=7.4 Hz, 3H).

Example 38: ethyl 2-(4-(4-(1-((tert-butoxycarbonyl)(methyl)amino)butyl)phenyl)-1H-pyrazol-1-yl)acetate

[0576] ##STR00195##

[0577] Prepared following general procedure 27. Obtained 56 mg, 76% yield.

[0578] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.83 (s, 1H), 7.74 (s, 1H), 7.49-7.44 (m, 2H), 7.33-7.28 (m, 2H), 5.48-5.19 (m, 1H), 4.95 (s, 2H), 4.28 (q, 2H, J=7.2 Hz), 2.59 (s, 3H), 1.90 (m, 2H), 1.52 (s, 9H), 1.44-1.35 (m, 2H), 1.32 (t, 3H, J=7.2 Hz), 1.02 (t, 3H, J=7.3 Hz).

Example 39: methyl 4-(1-((tert-butoxycarbonyl)(methyl)amino)butyl)-[1,1-biphenyl]-4-carboxylate

[0579] ##STR00196##

[0580] Prepared following general procedure 27. Obtained 66 mg, 87% yield.

[0581] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 8.12 (m, 2H), 7.67 (m, 2H), 7.61 (m, 2H), 7.40 (m, 2H), 5.50-5.19 (m, 1H), 3.96 (s, 3H), 2.62 (s, 3H), 1.92 (m, 2H), 1.53 (s, 9H), 1.47-1.33 (m, 2H), 1.02 (t, 3H, J=7.3 Hz).

Example 40: ethyl 3-(4-(1-((tert-butoxycarbonyl)(methyl)amino)butyl)phenyl)propanoate

[0582] ##STR00197##

[0583] Prepared following general procedure 28. Obtained 4.81 g, 73.2% yield.

[0584] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.20 (d, J=8.0 Hz, 2H), 7.15 (d, J=8.2 Hz, 2H), 5.46-5.13 (m, 1H), 4.12 (q, J=7.2 Hz, 2H), 2.93 (t, J=7.8 Hz, 2H), 2.61 (t, J=7.8 Hz, 2H), 2.53 (s, 3H), 1.83 (q, J=7.7, 6.8 Hz, 2H), 1.48 (s, 9H), 1.38-1.30 (m, 2H), 1.23 (t, J=7.1 Hz, 3H), 0.98 (t, J=7.4 Hz, 3H).

Example 41: 3-(4-(1-((tert-butoxycarbonyl)(methyl)amino)butyl)phenyl)propanoic acid

[0585] ##STR00198##

[0586] To a stirred solution of ethyl 3-(4-(1-((tert-butoxycarbonyl)(methyl)amino) butyl)phenyl)propanoate (17 g, 46.8 mmol) in THF (200 ml) and EtOH (100 ml), sodium hydroxide (6.55 g, 164 mmol) (dissolved in Water (25 ml)) was added dropwise at 0? C. The reaction mixture was stirred at 40? C. for 5 h. The status of the reaction was monitored by UPLC. The reaction mixture was concentrated under reduced pressure and the residue was diluted with cold water. The aqueous portion was washed with n-hexane. Then the layers were separated and aqueous portion was acidified with aqueous potassium bisulphate. The aqueous portion was extracted with DCM and organic layer was concentrated under reduced pressure to 3-(4-(1-((tert-butoxycarbonyl)(methyl)amino)butyl)phenyl)propanoic acid (15 g, 40.2 mmol, 86% yield) as colorless oil. The obtained crude was taken to next step without further purification.

[0587] .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? (ppm)=12.08 (s, 1H), 7.40-7.04 (m, 4H), 5.32-4.91 (m, 1H), 3.27 (s, 3H), 2.85-2.76 (m, 2H), 2.56-2.52 (m, 2H), 1.81 (d, J=6.1 Hz, 2H), 1.42 (s, 9H), 1.33-1.20 (m, 2H), 0.95 (t, J=7.3 Hz, 3H). m/z: 334.2 [M+H].sup.+

Example 42: ethyl 3-(4-(1-((tert-butoxycarbonyl)(methyl)amino)butyl)phenyl)propanoate

[0588] ##STR00199##

[0589] Prepared following general procedure 8. Obtained 4.5 g, 100.0% yield.

[0590] .sup.1H-NMR (400 MHz, CDCl.sub.3): ? 12.15 (s, 1H), 7.42 (d, J=8.0 Hz, 2H), 7.3 (d, J=8.0 Hz, 2H), 4.08-0.08 (m, 1H), 3.59-3.40 (m, 3H), 2.85 (t, J=7.6 Hz, 2H), 2.30 (m, 2H), 2.61 (t, J=8.0 Hz, 2H), 1.84-1.98 (m, 2H), 1.12-1.05 (m, 2H), 0.83 (t, J=7.2 Hz, 3H). m/z: 236.3 [M+H].sup.+

Example 43: 3-(4-(1-(2-cyano-N-methylacetamido)butyl)phenyl)propanoic acid

[0591] ##STR00200##

[0592] Prepared following general procedure 9. Obtained 3.4 g, 60.9% yield.

[0593] .sup.1H-NMR (400 MHz, DMSO-d.sub.6): ? 12.15 (s, 1H), 7.27-7.20 (m, 4H), 5.59-5.57 (m, 1H), 3.87 (s, 2H), 2.82-2.78 (m, 2H), 2.68 (s, 3H), 2.51-2.50 (m, 2H), 1.84 (q, J=8.0 Hz, 2H), 1.30-1.27 (m, 2H), 0.90 (t, J=7.2 Hz, 3H): m/z: 301.2 [M+H].sup.+

Example 44: (E)-3-(4-(1-(2-cyano-N-methyl-3-(thiazol-2-yl)acrylamido)butyl)phenyl)propanoic acid

[0594] ##STR00201##

[0595] Prepared following general procedure 10. Obtained 4.2 g, 89% yield.

[0596] .sup.1H-NMR (400 MHz, CDCl.sub.3): ? 11.75 (s, 1H), 8.14 (dd, J=19.6, 3.2 Hz, 1H), 8.01-7.83 (m, 1H), 7.35-7.25 (m, 4H), 5.68-5.45 (m, 1H), 3.04-2.82 (m, 5H), 2.75 (s, 1H), 2.57-2.51 (m, 2H), 2.05-1.99 (m, 2H), 1.38-1.36 (m, 2H), 0.98 (t, J=7.2 Hz, 3H). m/z: 398.1 [M+H]

Example 45: 4-(pyrrolidin-2-yl)phenol hydrobromide

[0597] ##STR00202##

[0598] Prepared following general procedure 4 from commercially available 2-(4-methoxyphenyl)pyrrolidine. Obtained 184 mg, 66.8% yield. m/z=164.3 ((M+H).sup.+

Example 46: 4-(piperidin-2-yl)phenol hydrobromide

[0599] ##STR00203##

[0600] Prepared following general procedure 4 from commercially available 2-(4-methoxyphenyl)piperidine. Obtained 38.4 mg, 28.5% yield. m/z=178.3 ((M+H).sup.+

Example 47: tert-butyl 2-(4-hydroxyphenyl)pyrrolidine-1-carboxylate

[0601] ##STR00204##

[0602] Prepared following general procedure 5 from 4-(pyrrolidin-2-yl)phenol hydrobromide. Obtained 142 mg, 71.5% yield.

[0603] .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.18 (s, 1H), 6.94 (d, J=8.0 Hz, 2H), 6.68 (d, J=8.3 Hz, 2H), 4.75-4.56 (m, 1H), 3.51-3.38 (m, 2H), 2.26-2.12 (m, 1H), 1.87-1.72 (m, 2H), 1.71-1.61 (m, 1H), 1.43-1.06 (m, 9H).

Example 48: tert-butyl 2-(4-hydroxyphenyl)piperidine-1-carboxylate

[0604] ##STR00205##

[0605] Prepared following general procedure 5 from 4-(piperidin-2-yl)phenol hydrobromide. Obtained 34.6 mg, 83.9%.

[0606] m/z=222.2 ((M-t-Bu)+H).sup.+

Example 49: tert-butyl 2-(4-(2-ethoxy-2-oxoethoxy)phenyl)pyrrolidine-1-carboxylate

[0607] ##STR00206##

[0608] Prepared following general procedure 6. Obtained 155 mg, 82.3% yield.

[0609] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.13-7.04 (m, 2H), 6.88-6.80 (m, 2H), 4.87-4.72 (m, 1H), 4.59 (s, 2H), 4.27 (q, J=7.1 Hz, 2H), 3.66-3.50 (m, 2H), 2.32-2.20 (m, 1H), 1.95-1.73 (m, 3H), 1.41-1.08 (m, 12H).

Example 50: tert-butyl 2-(4-(2-ethoxy-2-oxoethoxy)phenyl)piperidine-1-carboxylate

[0610] ##STR00207##

[0611] Prepared following general procedure 6. Obtained 34.7 mg, 76.8% yield.

[0612] m/z=308.3 ((M-t-Bu)+H).sup.+

Example 51: 2-(4-(1-(tert-butoxycarbonyl)pyrrolidin-2-yl)phenoxy)acetic acid

[0613] ##STR00208##

[0614] Prepared following general procedure 7. Obtained 71.3 mg, 95% yield. m/z=266.2 ((M-t-Bu)+H).sup.+

Example 52: 2-(4-(1-(tert-butoxycarbonyl)piperidin-2-yl)phenoxy)acetic acid

[0615] ##STR00209##

[0616] Prepared following general procedure 7. Obtained 31.0 mg, 96.5% yield. m/z=280.2 ((M-t-Bu)+H).sup.+

Example 53: tert-butyl 6-hydroxy-3,4-dihydroquinoline-1(2H)-carboxylate

[0617] ##STR00210##

[0618] Prepared following general procedure 5 from commercially available 1,2,3,4-tetrahydroquinolin-6-ol. Obtained 662 mg, 79.2% yield.

[0619] .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.06 (s, 1H), 7.29 (d, J=8.8 Hz, 1H), 6.51 (dd, J=8.8, 2.5 Hz 1H), 6.47 (d, J=2.5 Hz, 1H), 3.59-3.51 (m, 2H), 2.62 (t, J=6.6 Hz, 2H), 1.83-1.72 (m, 2H), 1.43 (s, 9H), m/z=194.2 ((M-tBu)+H).sup.+.

Example 54: tert-butyl 6-(2-(tert-butoxy)-2-oxoethoxy)-3,4-dihydroquinoline-1(2H)-carboxylate

[0620] ##STR00211##

[0621] Prepared following general procedure 6. Obtained 782 mg, 81.3% yield.

[0622] .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.54 (d, J=9.0 Hz, 1H), 6.69 (dd, J=9.0, 3.0 Hz, 1H), 6.61 (d, J=3.0 Hz, 1H), 4.46 (s, 2H), 3.71-3.63 (m, 2H), 2.72 (t, J=6.6 Hz, 2H), 1.95-1.84 (m, 2H), 1.50 (s, 9H), 1.49 (s, 9H), m/z=252.2 ((M?2tBu)+H).sup.+.

Example 55: 2-((1,2,3,4-tetrahydroquinolin-6-yl)oxy)acetic acid

[0623] ##STR00212##

[0624] Prepared following general procedure 33. Obtained 489 mg, 95.0% yield. m/z=208.2 (M+H).sup.+.

Example 56: 2-((1-(2-cyanoacetyl)-1,2,3,4-tetrahydroquinolin-6-yl)oxy)acetic acid

[0625] ##STR00213##

[0626] Prepared following general procedure 9. Obtained 174 mg, 68.2% yield. m/z=275.2 (M+H).sup.+.

Example 57: (E)-2-((1-(2-cyano-3-(thiazol-2-yl)acryloyl)-1,2,3,4-tetrahydroquinolin-6-yl)oxy)acetic acid

[0627] ##STR00214##

[0628] Prepared following general procedure 9. Obtained 73 mg, 47.1% yield

[0629] .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 12.96 (s, 1H), 8.21-8.16 (m, 2H), 8.11 (s, 1H), 7.16 (d, J=8.8 Hz, 1H), 6.86 (d, J=2.9 Hz, 1H), 6.72 (dd, J=8.8, 2.9 Hz, 1H), 4.65 (s, 2H), 3.76 (t, J=6.6 Hz, 2H), 2.72 (t, J=6.6 Hz, 2H), 1.94 (p, J=6.6 Hz, 2H); m/z=370.3 (M+H).sup.+.

Example 58: 4-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(17-amino-3,6,9,12,15-pentaoxaheptadecyl)benzamide-hydrochloride salt

[0630] ##STR00215##

[0631] To a solution of tert-butyl (17-amino-3,6,9,12,15-pentaoxaheptadecyl)carbamate (38.5 mg, 1.1 equiv., 101 ?mol) in DMF (1 mL), iBET726 (40.0 mg, 1.0 equiv., 92.0 ?mol), HATU (35.0 mg, 1.0 equiv., 92.0 ?mol) and N-ethyl-N-isopropylpropan-2-amine (35.7 mg, 48.1 ?L, 3.0 equiv., 276 ?mol) in DMF (1 mL) were added. The mixture was stirred at rt for 15 min. The crude was diluted in EtOAc and washed with water and brine. Purification by flash chromatography (DCM/MeOH (0-15%)) yielded the desired product as a yellow oil, 74 mg (quantitative yield).

[0632] The oil was dissolved in DCM (1 mL) and treated with HCl (1.49 g, 1.00 mL, 4 M, 40 equiv., 4.00 mmol) and the resulting heterogenous mixture was stirred at rt for 1 h. The reaction mixture was directly evaporated and dried at high vacuum to deliver the desired product as HCl salt (70 mg, 98%) as a colourless solid, m/z=697.7 (M+H).sup.+.

Example A1: 4-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1-(4-(1-((E)-2-cyano-3-(thiazol-2-yl)acrylamido)butyl)phenoxy)-2-oxo-6,9,12,15,18-pentaoxa-3-azaicosan-20-yl)benzamide

[0633] ##STR00216##

[0634] A solution of preparative compound 58 (5.00 mg, 1.0 equiv., 7.17 ?mol) in DMF (0.3 mL) was treated with a solution of 2-(4-(1-(2-cyano-3-(thiazol-2-yl)acrylamido)butyl)phenoxy)acetic acid (2.76 mg, 1.0 equiv., 7.17 ?mol) in DMF (0.3 mL), HATU (5.45 mg, 2.0 equiv., 14.3 ?mol) and DIPEA (2.78 mg, 3.75 ?L, 3.0 equiv., 21.5 ?mol) in DMF (0.3 mL). The mixture was stirred at rt for 15 min. The crude mixture was diluted to 1 mL with MeOH and directly purified using preparative HPLC (using a gradient from 5% to 95% of acetonitrile in water containing 0.1% of formic acid over 10 minutes) to deliver the product A1 (5.0 mg, 4.7 ?mol, 65%) as a pale yellow wax; m/z=1064.4 (M+H).sup.+.

Example A2: 4-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1-(4-(1-((E)-2-cyano-N-methyl-3-(thiazol-2-yl)acrylamido)butyl)phenoxy)-2-oxo-6,9,12,15,18-pentaoxa-3-azaicosan-20-yl)benzamide

[0635] ##STR00217##

[0636] In a test tube preparative compound 58 (8.0 mg, 1.1 equiv., 11 ?mol) was dissolved in 100 ?L of DMF. 15 ?L of DIPEA was added and the tube was mixed. (E)-2-(4-(1-(2-cyano-N-methyl-3-(thiazol-2-yl)acrylamido)butyl)phenoxy)acetic acid (4.0 mg, 1.0 equiv., 10 ?mol) was dissolved in DMF (0.3 mL). 15 ?L of DIPEA was added, followed by HATU (4.6 mg, 1.2 equiv., 12 ?mol). This was mixed and added to the tube containing the amine solution. The tube was shaken and left for around 1 hr. LCMS showed consumption of the starting materials and formation of the desired product. 0.5 mL of MeOH was added and the product was purified on preparative HPLC HPLC (using a gradient from 20% to 95% of acetonitrile in water containing 0.1% of formic acid over 10 minutes) to give the purified product, 2.7 mg (26% yield), m/z=1079.0 (M+H).sup.+.

Additional Examples

[0637]

TABLE-US-00006 Example m/z number Structure/Preparation (M + H).sup.+ A3 [00218] embedded image Prepared as described for example A2 1048.4 A5 [00219] Prepared as described for example A2 1106.5 A6 [00220] Prepared as described for example A2 1053.5 A40 [00221] Prepared as described for example A2 1075.2 A41 [00222] Prepared as described for example A2 1049.1

Example A4: 4-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1-(4-(1-(2-cyano-N-methyl-3-(thiazol-2-yl)propanamido)butyl)phenoxy)-2-oxo-6,9,12,15,18-pentaoxa-3-azaicosan-20-yl)benzamide

[0638] ##STR00223##

[0639] A solution preparative compound A2 (13.7 mg, 1.0 equiv., 12.7 ?mol) in THF (1 mL) was treated sodium triacetoxyborohydride (13.5 mg, 5.0 equiv., 63.5 ?mol) and stirred at 50? C. overnight. The reaction was quenched by addition of NH.sub.4Cl and extracted with CH.sub.2Cl.sub.2. The combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by preparative HPLC (using a gradient from 5% to 95% of acetonitrile in water containing 0.1% of formic acid over 10 minutes) yielded the desired product as a colourless solid, m/z=1080.5 (M+H).sup.+.

Example 59: tert-butyl 2-(4-((1-(4-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)-1,21-dioxo-5,8,11,14,17-pentaoxa-2,20-diazadocosan-22-yl)oxy)phenyl)pyrrolidine-1-carboxylate

[0640] ##STR00224##

[0641] A solution of 4-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(17-amino-3,6,9,12,15-pentaoxaheptadecyl)benzamide, HCl (20.5 mg, 280 ?L, 0.1 M, 1.0 equiv., 28.0 ?mol) in DMF (1 mL) was treated sequentially with a solution of 2-(4-(1-(tert-butoxycarbonyl)pyrrolidin-2-yl)phenoxy)acetic acid (9.00 mg, 280 ?L, 0.1 M, 1.0 equiv., 28.0 ?mol), 2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethylisouronium hexafluorophosphate(V) (10.6 mg, 1.0 equiv., 28.0 ?mol) and N-ethyl-N-isopropylpropan-2-amine (10.9 mg, 14.6 ?L, 3.0 equiv., 84.0 ?mol). The mixture was stirred at rt for 15 min. LC-MS showed complete conversion to a new product with the expected mass. The reaction was diluted with CH.sub.2Cl.sub.2 (5 mL) and quenched with H.sub.2O (2 mL). The phases were separated, and the organic layer was washed with H.sub.2O (2 mL) and NaHCO.sub.3 (2 mL). Purification by flash chromatography (CH.sub.2Cl.sub.2/MeOH) yielded the desired product (27.3 mg, 27.3 ?mol, 97.4%) as a yellow oil. m/z=1000.5 (M+H).sup.+.

Additional Examples

[0642]

TABLE-US-00007 Example m/z number Structure/Preparation (M + H).sup.+ 60 [00225] embedded image Prepared as described for example 59 1000.4 60a [00226] Prepared as described for example 59 1028.5 60b [00227] Prepared as described for example 59 using commercially available 2-(tert-butoxycarbonyl)-2,3,4,5-tetrahydro-1H- benzo[c]azepine-8-carboxylic acid [M ? H].sup.+ 969.4 60c [00228] embedded image Prepared as described for example 59 1015.1

Example 61: 4-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1-(4-(1-(2-cyanoacetyl)pyrrolidin-2-yl)phenoxy)-2-oxo-6,9,12,15,18-pentaoxa-3-azaicosan-20-yl)benzamide

[0643] ##STR00229##

[0644] To a solution of tert-butyl 2-(4-((1-(4-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)phenyl)-1,21-dioxo-5,8,11,14,17-pentaoxa-2,20-diazadocosan-22-yl)oxy)phenyl)pyrrolidine-1-carboxylate (27 mg, 1.0 equiv., 27 ?mol), in DCM (2 mL), HCl (4M in dioxane, 340 ?L, 50 equiv., 1.38 mmol) was added. The mixture was stirred for one hour at r.t., then evaporated to dryness. The crude was dissolved in dioxane (2 mL) and treated with 3-(3,5-dimethyl-1H-pyrazol-1-yl)-3-oxopropanenitrile (4.9 mg, 1.1 equiv., 30 ?mol) and triethylamine (11.4 ?L, 3.0 equiv., 82 ?mol) at 90? C. for 2 hours. Complete conversion to a new product was observed. Volatiles were evaporated under reduced pressure and the crude was purified by flash chromatography (CH.sub.2Cl.sub.2/MeOH) to obtain the desired product (16.3 mg, 61.7%), m/z=968.1 (M+H).sup.+.

Additional Examples

[0645]

TABLE-US-00008 Example m/z number Structure/Preparation (M + H).sup.+ 62 [00230] embedded image Prepared as described for example 61 967.5 63 [00231] Prepared as described for example 61 995.5 64 [00232] Prepared as described for example 61 937.2 65 [00233] Prepared as described for example 61 982.1

Example A7: 4-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1-(4-(1-((E)-2-cyano-3-(thiazol-2-yl)acryloyl)pyrrolidin-2-yl)phenoxy)-2-oxo-6,9,12,15,18-pentaoxa-3-azaicosan-20-yl)benzamide

[0646] ##STR00234##

[0647] A solution of 4-((2S,4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-N-(1-(4-(1-(2-cyanoacetyl)pyrrolidin-2-yl)phenoxy)-2-oxo-6,9,12,15,18-pentaoxa-3-azaicosan-20-yl)benzamide (16.3 mg, 1.0 equiv., 16.8 ?mol) in dioxane (2 mL) was treated with thiazole-2-carbaldehyde (3.7 ?L, 2.5 equiv., 42.1 ?mol) and piperidine (1.66 ?L, 1.0 equiv., 16.8 ?mol) and the mixture was heated to 66? C. for 72 h. The volatiles were evaporated under reduced pressure. Purification by reverse phase HPLC (using a gradient from 5% to 95% of acetonitrile in water containing 0.1% of formic acid over 10 minutes) gave the desired product (8 mg, 45% yield). m/z=1063.7 (M+H).sup.+.

Additional Examples

[0648]

TABLE-US-00009 Example m/z number Structure/Preparation (M + H).sup.+ A8 [00235] embedded image Prepared as described for example A7 1062.2 A9 [00236] Prepared as described for example A7 1090.5 A42 [00237] Prepared as described for example A7 1032.3 A43 [00238] Prepared as described for example A7 1077.2

Example 66: tert-butyl 4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl) thio)thiazol-2-yl)carbamoyl)-[1,4-bipiperidine]-1-carboxylate

[0649] ##STR00239##

[0650] SNS-032 (60 mg, 1.0 equiv., 0.16 mmol) and tert-butyl 4-oxopiperidine-1-carboxylate (47 mg, 1.5 equiv., 0.24 mmol) were suspended in DCM (1.5 mL). Tetraethoxytitanium (72 mg, 66 ?L, 2.0 equiv., 0.32 mmol) was added and all suspended solids went into solution. The reaction was stirred at room temperature overnight. Sodium cyanoborohydride (20 mg, 2.0 equiv., 0.32 mmol) was added and the reaction was stirred at room temperature for 2 hours. The reaction was quenched by addition of sat. NaHCO.sub.3 solution, filtered over celite and extracted three times with EtOAc. The organic layer was reduced in vacuo and dry loaded onto silica. The reaction was purified by flash chromatography (12 g column, 0 to 20% MeOH in DCM) to give tert-butyl 4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)carbamoyl)-[1,4-bipiperidine]-1-carboxylate (73 mg, 0.13 mmol, 82%) as a yellow solid. The Boc protected intermediate was then dissolved in DCM (2 ml) and treated with HCl (4 M in dioxane, 2 ml) for 1 hour. The reaction mixture was evaporated to dryness to obtain the desired product as hydrochloride salt in quantitative yield, m/z=464.2 (M+H).sup.+.

Additional Examples

[0651]

TABLE-US-00010 Example m/z number Structure Preparation (M + H).sup.+ 67 [00240] embedded image Prepared as described for example 66 490.2 68 [00241] Prepared as described for example 66 478.2 69 [00242] Prepared as described for example 66 464.2 70 [00243] Prepared as described for example 66 436.2 71 [00244] Prepared as described for example 66 450.2

Example A38: (E)-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(2-((2-(2-cyano-3-(thiazol-2-yl)acryloyl)-1-propyl-1,2,3,4-tetrahydroisoquinolin-6-yl)oxy)acetyl)piperidine-4-carboxamide

[0652] ##STR00245##

[0653] A solution of SNS032 (1.0 equiv.) in DMF (0.1 M) was treated with a solution of 36b (1.0 equiv.) HATU (1.1 equiv.) and DIPEA (2.5 equiv.) in DMF (0.1 M) and the reaction mixture was stirred at rt for 15 min. The reaction was quenched with water and extracted with EtOAc. The combined organic layers were washed with LiCl (5%), water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by preparative HPLC (using a gradient from 5% to 95% of acetonitrile in water containing 0.1% of formic acid over 10 minutes) yielded the desired product A38, m/z=774.3 (M+H).sup.+.

[0654] This reaction protocol is exemplified in relation to a THIQ analogue but is also applicable to the synthesis of N-alkylated analogues.

Additional Examples

[0655]

TABLE-US-00011 Exam- ple m/z number Structure Preparation (M + H).sup.+ A31 [00246] embedded image Prepared as described for example A38 869.4 A28 [00247] Prepared as described for example A38 843.4 A27 [00248] Prepared as described for example A38 857.4 A20 [00249] Prepared as described for example A38 873.4 A19 [00250] Prepared as described for example A38 845.3 A13 [00251] Prepared as described for example A38 817.3 A12 [00252] embedded image Prepared as described for example A38 845.3 A11 [00253] Prepared as described for example A38 831.3 A10 [00254] Prepared as described for example A38 762.3

Example A18: N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(2-(4-(1-(2-cyano-N-methyl-3-(thiazol-2-yl)propanamido)butyl)phenoxy)acetyl)-[1,4-bipiperidine]-4-carboxamide

[0656] ##STR00255##

[0657] Preparative compound A12 (14 mg, 1.0 equiv., 17 ?mol) was dissolved in THF (0.2 M). Sodium triacetoxyborohydride (11 mg, 3.0 equiv., 50 ?mol) was added and the reaction was stirred at rt for 5. A further portion of sodium triacetoxyborohydride (11 mg, 3.0 equiv., 50 ?mol) was added and the reaction was stirred at rt for 16 h. The reaction was diluted with water and extracted three times with CH.sub.2Cl.sub.2. Combined organic extracts were reduced in vacuo and purified by preparative HPLC (using a gradient from 5% to 95% of acetonitrile in water containing 0.1% of formic acid over 10 minutes) to furnish the desired product in 43% yield, m/z=848.2 (M+H).sup.+.

Additional Example

Example A30: N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(3-(4-(1-(2-cyano-N-methyl-3-(thiazol-2-yl)propanamido)butyl)phenyl)propanoyl)-[1,4-bipiperidine]-4-carboxamide

[0658] ##STR00256##

[0659] Prepared as described for compound A18 from preparative compound A28 in 74% yield,

[0660] m/z=845.4 (M+H).sup.+.

Example 72: tert-butyl 6-(2-oxoethoxy)-1-propyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0661] ##STR00257##

[0662] Prepared following general procedure 29. Obtained 97.6 mg, 65.0% yield. m/z=334.2 (M+H).sup.+.

Additional Examples

[0663]

TABLE-US-00012 Example m/z number Structure Preparation (M + H).sup.+ 73 [00258] embedded image Prepared following general procedure 29 306.2 74 [00259] Prepared following general procedure 29 322.2 75 [00260] Prepared following general procedure 29 372.2 76 [00261] Prepared following general procedure 29 342 [M + Na]

Example 77: tert-butyl 6-(2-(4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl) thio)thiazol-2-yl)carbamoyl)piperidin-1-yl)ethoxy)-1-propyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0664] ##STR00262##

[0665] SNS032 (55.9 mg, 1.0 equiv., 147 ?mol) and 72 (49.0 mg, 1.0 equiv., 147 ?mol) were suspended in THF (1.5 mL) and cooled to 0? C. sodium triacetoxyborohydride (62.3 mg, 2.0 equiv., 294 ?mol) was added and all suspended solids went into solution. The reaction was warmed to room temperature and left to stir overnight. The reaction was quenched by addition of NaHCO.sub.3 (sat. aq.) and extracted with CH.sub.2Cl.sub.2. The combined organic layers were washed with water and brine, dried over MgSO.sub.4 and concentrated under reduced pressure. Purification by flash chromatography yielded the corresponding amine. Obtained 76.7 mg, 75% yield. m/z=698.3 (M+H).sup.+.

[0666] This reaction protocol has been exemplified in relation to a THIQ analogue (such as those outlined in section 2.4 but is also applicable to the synthesis of N-alkylated analogues (such as those shown in 2.3).

Additional Examples

[0667]

TABLE-US-00013 Example m/z number Structure Preparation (M + H).sup.+ 78 [00263] embedded image Prepared as described for example 77 670.3 79 [00264] Prepared as described for example 77 684.4 80 [00265] Prepared as described for example 77 736.4

Example 81: tert-butyl (1-(4-(2-(4-((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)carbamoyl)-[1,4-bipiperidin]-yl)-2-oxoethoxy)phenyl)butyl) (methyl)carbamate

[0668] ##STR00266##

[0669] Compound 66 (49.0 mg, 1.0 equiv., 91 ?mol) and compound 41 (46.0 mg, 1.5 equiv., 140 ?mol) were coupled following general procedure 31. The product was purified by flash chromatography yielding 81. Obtained 71.0 mg, 95% yield. m/z=781.4 (M+H).sup.+.

TABLE-US-00014 Example m/z number Structure Preparation (M + H).sup.+ 82 [00267] embedded image Prepared as described for example 81 746.3 83 [00268] Prepared as described for example 81 829.4 84 [00269] Prepared as described for example 81 833.2 85 [00270] Prepared as described for example 81 750.3

Example A32: (E)-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(3-(4-(1-(N-methyl-3-(thiazol-2-yl)acrylamido)butyl)phenyl)propanoyl)-[1,4-bipiperidine]-4-carboxamide

[0670] ##STR00271##

[0671] Compound 81 (25.0 mg, 1.0 equiv., 32 ?mol) was dissolved in CH.sub.2Cl.sub.2 (1 mL). 4M HCl in dioxane (0.40 mL, 50 equiv., 1.6 mmol) was added and the reaction was stirred at room temperature for 1 hour. The reaction mixture was reduced in vacuo to give the unprotected product. This was then coupled with compound 19 (7.4 mg, 1.5 equiv., 47 ?mol) following general procedure 31. The product was purified by preparative HPLC (using a gradient from 5% to 95% of acetonitrile in water containing 0.1% of formic acid over 10 minutes) to furnish A32. Obtained 12.0 mg, 46% yield. m/z=818.3 (M+H).sup.+.

Example 86: N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(2-((2-(2-cyanoacetyl)-1-propyl-1,2,3,4-tetrahydroisoquinolin-6-yl)oxy)ethyl)piperidine-4-carboxamide

[0672] ##STR00272##

[0673] A solution of Compound 77 (76.7 mg, 1.0 equiv., 110 ?mol) in CH.sub.2Cl.sub.2 (0.05 M) was treated with HCl (4 M in dioxane, 1.37 mL, 50 equiv., 5.5 mmol)) and the mixture was stirred for 2 h. The volatiles were evaporated under reduced pressure. The residue was dissolved in 1,4-dioxane (0.05 M), treated with Et.sub.3N (80 ?L, 4.0 equiv., 440 ?mol) and 3-(3,5-dimethyl-1H-pyrazol-1-yl)-3-oxopropanenitrile (19.7 mg, 1.1 equiv., 121 ?mol) and the mixture was heated to 90? C. for 2 h. The volatiles were evaporated under reduced pressure. Purification by flash chromatography yielded the desired product (56.8 mg, 77.7% yield). m/z=665.3 (M+H).sup.+.

[0674] This reaction protocol has been exemplified in relation to a THIQ analogue (such as those outlined in section 2.4 but is also applicable to the synthesis of N-alkylated analogues (such as those shown in 2.3).

Additional Examples

[0675]

TABLE-US-00015 Example m/z number Structure Preparation (M + H).sup.+ 87 [00273] embedded image Prepared as described for example 86 637.3 88 [00274] Prepared as described for example 86 651.3 89 [00275] Prepared as described for example 86 703.3 90 [00276] Prepared as described for example 86 713.2 91 [00277] Prepared as described for example 86 796.5 92 [00278] Prepared as described for example 86 800.4 93 [00279] embedded image Prepared as described for example 86 717.3

Example A35: (E)-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(2-((2-(2-cyano-3-(thiazol-2-yl)acryloyl)-1-propyl-1,2,3,4-tetrahydroisoquinolin-6-yl)oxy)ethyl)piperidine-4-carboxamide

[0676] ##STR00280##

[0677] A solution of compound 86 (35 mg, 1.0 equiv., 52.6 ?mol) in THF (0.1 M) was treated with thiazole-2-carbaldehyde (28 mg, 5.0 equiv., 263 ?mol) and piperidine (105 ?L, 2.0 equiv., 105 ?mol) and the mixture was heated to 66? C. for 72 h. The volatiles were evaporated under reduced pressure. Purification by flash chromatography yielded A35 (20.9 mg, 52.2% yield). m/z=760.3 (M+H).sup.+.

[0678] Again, while these general reaction protocols have been exemplified in relation to a THIQ analogue (such as those outlined in section 2.6, it is also applicable to the synthesis of N-alkylated analogues (such as those shown in 2.5).

Additional Examples

[0679]

TABLE-US-00016 Example m/z number Structure Preparation (M + H).sup.+ A37 [00281] embedded image Prepared as described for example A35 832.2 A36 [00282] Prepared as described for example A35 782.3 A34 [00283] Prepared as described for example A35 732.2 A33 [00284] Prepared as described for example A35 739.3 A29 [00285] Prepared as described for example A35 798.3 A26 [00286] Prepared as described for example A35 719.4 A25 [00287] embedded image Prepared as described for example A35 796.3 A24 [00288] Prepared as described for example A35 746.3 A23 [00289] Prepared as described for example A35 808.3 A22 [00290] Prepared as described for example A35 818.3 A21 [00291] Prepared as described for example A35 758.3 A44 [00292] Prepared as described for example A35 777.4 A45 [00293] embedded image Prepared as described for example A35 783.3 A15 [00294] Prepared as described for example A35 808.4 A17 [00295] Prepared as described for example A35 891.3 A16 [00296] Prepared as described for example A35 895.4 A14 [00297] Prepared as described for example A35 812.3

Example A46: N-[5-[(5-tert-butyloxazol-2-yl)methylsulfanyl]thiazol-2-yl]-1-[2-[4-[4-[1-[[(E)-2-cyano-3-(1-methylimidazol-2-yl)prop-2-enoyl]-methyl-amino]butyl]phenyl]pyrazol-1-yl]acetyl]piperidine-4-carboxamide

[0680] ##STR00298##

[0681] A solution of example 93 (12 mg, 1.0 equiv., 17 ?mol) in EtOH (0.01M) was treated with 1-methyl-2-imidazolecarboxaldehyde (3.7 mg, 2.0 equiv., 34 ?mol) and piperidine (10 ?L, 2.0 equiv., 34 ?mol) and the mixture was stirred at room temperature for 18 h. The volatiles were evaporated under reduced pressure. Purification by preparative HPLC (using a gradient from 5% to 95% of acetonitrile in water containing 0.1% of formic acid over 10 minutes) yielded the corresponding product (5.4 mg, 13.8% yield). m/z=810.3 (M+H).sup.+

TABLE-US-00017 Example |m/z number Structure Preparation (M + H).sup.+ A47 [00299] embedded image Prepared as described for example A46 810.3 A48 [00300] Prepared as described for example A46 824.3 A49 [00301] Prepared as described for example A46 796.4 A50 [00302] Prepared as described for example A46 871.4 A51 [00303] Prepared as described for example A46 846.2 A52 [00304] Prepared as described for example A46 873.2

Example 945-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine

[0682] ##STR00305##

[0683] In a screw-top vialSNS032 (120 mg, 1 equiv., 315 ?mol) was dissolved in HCl (2.63 mL, 6 molar) and heated at 90? C. for 3 hours. LCMS indicates formation of desired product (retention time=1.74 min, M+H=270). This was neutralised with sat. NaHCO.sub.3 and extracted three times with DCM. Combined organic were dried over MgSO.sub.4 and reduced in vacuo to give the desired product as a off-white solid in quantitative yield. m/z=270.1 [m+H.sup.+].

Example 95tert-butyl 4-(((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)methyl)piperidine-1-carboxylate

[0684] ##STR00306##

[0685] Compound 94 (52 mg, 1 Eq, 0.19 mmol) and tert-butyl 4-formylpiperidine-1-carboxylate (62 mg, 1.5 Eq, 0.29 mmol) were suspended in THF (2.5 mL) and cooled to 0? C. Sodium triacetoxyborohydride (82 mg, 2 Eq, 0.39 mmol) and titanium (IV) ethoxide (0.14 g, 0.12 mL, 65% Wt, 2 Eq, 0.39 mmol) were added and the reaction left to stir for 4 hours. A further portion of sodium triacetoxyborohydride (82 mg, 2 Eq, 0.39 mmol) was added and left to stir for 1 hour. LCMS indicates complete formation of desired product (retention time=3.25 min, M+H=467). The reaction was quenched by addition of sat. NaHCO.sub.3 solution and left to stir for 15 mins. The reaction was filtered through a pad of Celite, washing with DCM. The filtrate was reduced in vacuo and purified by flash chromatography (4 g column, 0 to 20% MeOH in DCM) to give the desired product (90 mg, 0.19 mmol, 100%) as a yellow oil. m/z=467.2 [m+H.sup.+].

Example 965-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-N-(piperidin-4-ylmethyl)thiazol-2-amine (hydrochloride salt)

[0686] ##STR00307##

[0687] Compound 95 (90 mg, 1 Eq, 0.19 mmol) was dissolved in DCM (2.5 mL), HCl in dioxane (0.35 g, 2.4 mL, 4 molar, 50 Eq, 9.6 mmol) was added and the reaction was stirred at room temperature for 1 hour. LCMS indicated consumption of the starting material and formation of the desired product (retention time=0.31 min, M+H=367). The reaction was reduced in vacuo and dried under vacuum overnight to give the desired product (85 mg, 0.19 mmol, 100%) as a yellow solid. m/z=367.2 [m+H.sup.+].

Example 97tert-butyl (1-(4-(3-(4-(((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)methyl)piperidin-1-yl)propyl)phenyl)butyl)(methyl)carbamate

[0688] ##STR00308##

[0689] Compound 96 (85 mg, 1 Eq, 0.19 mmol) and compound 76 (0.11 g, 1.8 Eq, 0.35 mmol) were dissolved in THF (3 mL). Sodium triacetoxyborohydride (82 mg, 2 Eq, 0.39 mmol) was added and the reaction was left to stir for 4 hours. LCMS showed formation of desired product (retention time=3.08 min, M+H=670). The reaction was diluted with water and extracted three times with DCM. Combined organic extracts were reduced in vacuo and dry loaded onto silica. The reaction was purified by flash chromatography (12 g column, 0 to 20% MeOH in DCM) to give the desired product (33 mg, 49 ?mol, 25%) as a yellow oil. m/z=670.3 [m+H.sup.+].

Example 985-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-N-((1-(3-(4-(1-(methylamino)butyl)phenyl)propyl)piperidin-4-yl)methyl)thiazol-2-amine (hydrochloride salt)

[0690] ##STR00309##

[0691] Compound 97 (33 mg, 1 Eq, 49 ?mol) was dissolved in CH.sub.2Cl.sub.2 (2.5 mL). HCl in dioxane (90 mg, 0.62 mL, 4 molar, 50 Eq, 2.5 mmol) was added and the reaction was stirred at room temperature for 1 hour. LCMS indicated consumption of starting material and formation of desired product (retention time=0.28 min, M+H=570). The reaction was reduced in vacuo and dried under vacuum overnight to give the desired product (32 mg, 50 ?mol, 100%) as a white solid. m/z=570.3 [m+H.sup.+].

Example A393-(benzo[d]thiazol-2-yl)-N-(1-(4-(3-(4-(((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)methyl)piperidin-1-yl)propyl)phenyl)butyl)-2-cyano-N-methylacrylamide

[0692] ##STR00310##

[0693] Compound 98 (32 mg, 1 Eq, 50 ?mol) was suspended in 1,4-Dioxane (3 mL). 3-(3,5-dimethyl-1H-pyrazol-1-yl)-3-oxopropanenitrile (9.7 mg, 1.2 Eq, 60 ?mol) and DIPEA (19 mg, 26 ?L, 3 Eq, 0.15 mmol) were added and the reaction was heated at 90? C. for 4 hours. LCMS indicated formation of acylated product (retention time=2.50 min, M+H=637). The reaction was reduced in vacuo and purified by flash chromatography (4 g column, 0 to 20% MeOH in DCM) to give A39 (30 mg, 47 ?mol, 95%). This was dissolved in THF (2.5 mL) and transferred to a screw-top vial. Benzo[d]thiazole-2-carbaldehyde (20 mg, 2.5 Eq, 0.12 mmol) and piperidine (2.1 mg, 2.5 ?L, 0.5 Eq, 25 ?mol) were added, the vial was capped and heated at 70? C. overnight. LCMS indicates formation of desired product (retention time=2.95 min, M+H=782two peaks present in a 1:1 ratio (E/Z isomers)). A further benzo[d]thiazole-2-carbaldehyde (20 mg, 2.5 Eq, 0.12 mmol) and piperidine (2.1 mg, 2.5 ?L, 0.5 Eq, 25 ?mol) were added and the reaction heated at 70? C. overnight. LCMS indicated complete consumption of starting material. The reaction mixture was reduced in vacuo and dissolved in 2 mL of MeOH. The product was purified by preparative HPLC (eluting from 20% to 95% of MeCN in H.sub.2O+0.1% formic acid) to give the desired product (14 mg, 18 ?mol, 35%, 98% purity). m/z=782.3 [m+H.sup.+].

Example 99: N-(1-(3-bromophenyl)propan-2-yl)acetamide

[0694] ##STR00311##

[0695] Prepared following general procedure 34. Obtained 809 mg, 49.9% yield, m/z=256.2 (M+H).sup.+.

Example 100: 6-bromo-1,3-dimethyl-3,4-dihydroisoquinoline

[0696] ##STR00312##

[0697] Prepared following general procedure 35. Obtained 526 mg, 69.9% yield, m/z=238.1 (M+H).sup.+.

Example 101: 6-bromo-1,3-dimethyl-1,2,3,4-tetrahydroisoquinoline

[0698] ##STR00313##

[0699] Prepared following general procedure 3. Obtained 460 mg, 86.7% yield, m/z=240.1 (M+H).sup.+.

Example 102: tert-butyl 6-bromo-1,3-dimethyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0700] ##STR00314##

[0701] Prepared following general procedure 5. Obtained 486 mg, 74.6% yield, m/z=284.2 ((M-tBu)+H).sup.+.

Example 103: tert-butyl (E)-6-(3-ethoxy-3-oxoprop-1-en-1-yl)-1,3-dimethyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0702] ##STR00315##

[0703] Prepared following general procedure 27. Obtained 313 mg, 60.9% yield, m/z=304.3 ((M-tBu)+H).sup.+.

Example 104: tert-butyl 6-(3-ethoxy-3-oxopropyl)-1,3-dimethyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0704] ##STR00316##

[0705] Prepared following general procedure 28. Obtained 311 mg, 98.5% yield, m/z=362.4 ((M)+H).sup.+.

Example 105: 3-(2-(tert-butoxycarbonyl)-1,3-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-yl)propanoic acid

[0706] ##STR00317##

[0707] Prepared following general procedure 7. Obtained 120 mg, 86.7% yield, m/z=362.4 ((M)+H).sup.+.

Example 106: 3-(1,3-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-yl)propanoic acid

[0708] ##STR00318##

[0709] Prepared following general procedure 8. Obtained 70.1 mg, 72.1% yield, m/z=234.2 ((M)+H).sup.+.

Example 107: 3-(2-(2-cyanoacetyl)-1,3-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-yl)propanoic acid

[0710] ##STR00319##

[0711] Prepared following general procedure 9. Obtained 70.1 mg, 72.1% yield, m/z=301.3 ((M)+H).sup.+.

Example 108: (E)-3-(2-(2-cyano-3-(thiazol-2-yl)acryloyl)-1,3-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-yl)propanoic acid

[0712] ##STR00320##

[0713] Prepared following general procedure 10. Obtained 35.7 mg, 43.2% yield, m/z=396.3 ((M)+H).sup.+.

Example 109N-(2-bromophenethyl)acetamide

[0714] ##STR00321##

[0715] Prepared following general procedure 1. Obtained 60 g, 99% yield, m/z=242.1 [M+H].sup.+ 244.1 [M+H].sup.+, 1H NMR (400 MHz, DMSO-d.sub.6): 7.96 (s, 1H), 7.59 (d, J=7.6 Hz, 1H), 7.36-7.32 (m, 2H), 7.19-7.16 (m, 1H), 3.30-3.24 (m, 2H), 2.83 (t, J=6.8 Hz, 2H), 1.80 (s, 3H).

Example 1105-bromo-1-methyl-3,4-dihydroisoquinoline

[0716] ##STR00322##

[0717] Prepared following general procedure 35. Obtained 25 g, 45% yield, m/z=224.1 [M+H].sup.+ 226.0 [M+H].sup.+, .sup.1H NMR (400 MHz, DMSO-d.sub.6): 7.69 (d, J=0.8 Hz, 1H), 7.59 (d, J=7.6 Hz, 1H), 7.30 (t, J=8.0 Hz, 1H), 3.59-3.55 (m, 2H), 2.69 (t, J=7.2 Hz, 2H), 2.31 (s, 3H).

Example 1115-bromo-1-methyl-1,2,3,4-tetrahydroisoquinoline

[0718] ##STR00323##

[0719] Prepared following general procedure 3. Obtained 14 g, 93% yield, m/z=226.1 [M+H].sup.+ 228.2 [M+H].sup.+, .sup.1H NMR (400 MHz, DMSO-d.sub.6): 7.60 (d, J=6.8 Hz, 1H), 7.21-7.17 (m, 1H), 7.12 (t, J=8.0 Hz, 1H), 3.96-3.91 (m, 1H), 3.13-3.11 (m, 1H), 3.11-3.09 (m, 1H), 2.86-2.81 (m, 1H), 2.68-2.62 (m, 2H), 1.33 (d, J=6.8 Hz, 3H).

Example 112tert-butyl 5-bromo-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0720] ##STR00324##

[0721] Prepared following general procedure 5. Obtained 15 g, 72% yield, m/z=226.1 [M?100+H].sup.+228.0 [M?100+H].sup.+, .sup.1H NMR (400 MHz, DMSO-d.sub.6): 7.49 (d, J=7.2 Hz, 1H), 7.29 (d, J=7.6 Hz, 1H), 7.16 (t, J=7.6 Hz, 1H), 5.12 (s, 1H), 4.04 (s, 1H), 3.21-3.16 (m, 1H), 2.77 (m, 1H), 2.63-2.59 (m, 1H), 1.47 (m, 12H).

Example 113N-(3-bromophenethyl)acetamide

[0722] ##STR00325##

[0723] Prepared following general procedure 1. Obtained 60 g, 95% yield, .sup.1H NMR (400 MHz, DMSO-d.sub.6): 7.91 (s, 1H), 7.42-7.39 (m, 1H), 7.28-7.21 (m, 1H), 3.27 (q, J=7.2 Hz, 2H), 2.70 (t, J=7.2 Hz, 2H), 1.78 (s, 3H).

Example 1146-bromo-1-methyl-3,4-dihydroisoquinoline

[0724] ##STR00326##

[0725] Prepared following general procedure 35. Obtained 37 g, 64% yield, m/z=224.0 [M+H].sup.+ 225.9 [M+H].sup.+, .sup.1H NMR (400 MHz, DMSO-d.sub.6): 7.55-7.47 (m, 3H), 3.52 (t, J=2.8 Hz, 2H), 2.65 (t, J=7.2 Hz, 2H), 2.29 (s, 3H).

Example 1156-bromo-1-methyl-1,2,3,4-tetrahydroisoquinoline

[0726] ##STR00327##

[0727] Prepared following general procedure 3. Obtained 25 g, 75% yield, m/z=226.1 [M+H].sup.+ 228.2 [M+H].sup.+, .sup.1H NMR (400 MHz, DMSO-d.sub.6): 7.30-7.26 (m, 2H), 7.11 (d, J=8.4 Hz, 1H), 3.88 (q, J=6.4 Hz, 1H), 3.33-3.04 (m, 1H), 2.79-2.71 (m, 3H), 2.59-2.51 (m, 1H), 1.31 (d, J=6.80 Hz, 3H).

Example 116tert-butyl 6-bromo-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0728] ##STR00328##

[0729] Prepared following general procedure 5. Obtained 12 g, 92% yield, m/z=226.1 [M?100+H].sup.+

Example 117N-(4-bromophenethyl)acetamide

[0730] ##STR00329##

[0731] Prepared following general procedure 1. Obtained 60 g, 96% yield, m/z=242.1 [M+H].sup.+, 244.1 [M+H].sup.+, .sup.1H NMR (400 MHz, DMSO-d.sub.6): 7.89 (s, 1H), 7.47 (d, J=2.4 Hz, 2H), 7.17 (d, J=2.4 Hz, 2H), 3.24 (q, J=5.6 Hz, 2H), 2.67 (t, J=7.2 Hz, 2H), 1.77 (s, 3H).

Example 1187-bromo-1-methyl-3,4-dihydroisoquinoline

[0732] ##STR00330##

[0733] Prepared following general procedure 35. Obtained 45 g, 81% yield, m/z=224.1 [M+H].sup.+ 226.0 [M+H].sup.+, .sup.1H NMR (400 MHz, DMSO-d.sub.6): 7.70 (s, 1H), 7.59 (d, J=6.0 Hz, 1H), 7.22 (d, J=8.0 Hz, 1H), 3.54-3.51 (m, 2H), 2.60 (t, J=7.2 Hz, 2H), 2.30 (s, 3H).

Example 1197-bromo-1-methyl-1,2,3,4-tetrahydroisoquinoline

[0734] ##STR00331##

[0735] Prepared following general procedure 3. .sup.1H NMR (400 MHz, DMSO-d.sub.6): 7.32 (s, 1H), 7.26 (d, J=0.4 Hz, 1H), 7.01 (d, J=8.4 Hz, 1H), 3.94-3.89 (m, 1H), 3.09-3.04 (m, 1H), 2.81-2.75 (m, 1H), 2.72-2.66 (m, 1H), 2.64-2.59 (m, 1H), 2.52-2.50 (m, 1H), 1.32 (d, J=6.8 Hz, 3H).

Example 120tert-butyl 7-bromo-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0736] ##STR00332##

[0737] Prepared following general procedure 5. Obtained 42 g, 92% yield, m/z=226.1 [M?100+H].sup.+228.1 [M?100+H].sup.+, .sup.1H NMR (400 MHz, DMSO-d.sub.6): 7.48 (s, 1H), 7.34 (d, J=2.0 Hz, 1H), 7.11 (d, J=8.4 Hz, 1H), 3.97-3.92 (m, 1H), 3.16-3.09 (m, 1H), 2.73-2.68 (m, 2H), 1.47-1.36 (m, 12H).

Example 121tert-butyl 6-(3-((tert-butyldimethylsilyl)oxy)prop-1-yn-1-yl)-1-methyl-3,4-dihydro isoquinoline-2(1H)-carboxylate

[0738] ##STR00333##

[0739] To a stirred solution of compound 116 (5 g, 1.0 eq, 15.33 mmol) and tert-butyldimethyl(prop-2-yn-1-yloxy)silane (3.92 g, 1.5 eq, 22.99 mmol) in DMF (50 ml) was added copper(I) iodide (0.876 g, 0.3 eq, 4.60 mmol), TEA (6.65 ml, 3.0 eq, 46.0 mmol) and the reaction mixture was degassed for 15 min with nitrogen. Pd(PPh.sub.3).sub.4 (1.77 g, 0.1 eq, 1.53 mmol) added to the reaction mixture which was stirred at 70? C. for 16 h. The reaction mixture was cooled to RT, quenched with ice-water, diluted with EtOAc and layers were separated. The aqueous layer was extracted with EtOAc and the combined organic layers were washed with brine, dried (Na.sub.2SO.sub.4) and filtered. The organic layers were concentrated in vacuo to give the crude product which was purified by flash chromatography (5 to 10% EtOAc in n-hexane) to give the desired product 121 (5 g, 12.03 mmol, 78% yield) as brown liquid.

[0740] .sup.1H NMR (400 MHz, CDCl.sub.3) 7.31-7.29 (m, 1H), 7.25-7.28 (m, 1H), 7.08-7.00 (m, 1H), 5.24-5.05 (m, 1H), 4.55 (s, 1H), 4.20-4.03 (m, 2H), 2.88-2.07 (m, 2H), 1.43-1.26 (m, 12H), 0.93 (s, 9H), 0.17 (s, 6H),

Example 122tert-butyl 6-(3-ethoxy-3-oxopropyl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0741] ##STR00334##

[0742] A solution of compound 116 (2.0 g, 1.0 equiv., 6.13 mmol) in THF (30 ml) was degassed with nitrogen for 10 min and Pd(tBu.sub.3P).sub.2 (0.31 g, 0.1 equiv., 0.61 mmol) was added and again degassed for 10 min. (3-ethoxy-3-oxopropyl)zinc(II) bromide (30.7 ml, 2.5 equiv., 15.33 mmol) was added to the reaction mixture and the reaction was stirred at RT for 16 h. The reaction was diluted with water and extracted with EtOAc. The combined organic layer was dried (Na.sub.2SO.sub.4), filtered and concentrated in vacuo. The crude compound was purified by flash chromatography (10-15% EtOAc in n-hexane) to isolate 122 (1.7 g, 4.89 mmol, 80% yield).

Example 1233-(2-(tert-butoxycarbonyl)-1-methyl-1,2,3,4-tetrahydroisoquinolin-6-yl)propanoic acid

[0743] ##STR00335##

[0744] Prepared following general procedure 7. Obtained 2.3 g, m/z=220.1 [M+H-100].sup.+

Example 124tert-butyl 6-(3-(methoxy(methyl)amino)-3-oxopropyl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0745] ##STR00336##

[0746] Prepared following general procedure 31. Crude material obtained 2.4 g, 77% yield, m/z 263.2 [M+H-100].sup.+

Example 125tert-butyl 1-methyl-6-(3-oxopropyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0747] ##STR00337##

[0748] Prepared following general procedure 32. Crude material obtained 0.53 g, 64% yield, m/z 204.2 [M+H-100].sup.+

Example 126tert-butyl (E)-7-(3-ethoxy-3-oxoprop-1-en-1-yl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0749] ##STR00338##

[0750] To a degassed solution of compound 120 (7 g, 1.0 equiv., 21.5 mmol) in DMF (50 ml) was added ethyl acrylate (4.57 ml, 2.0 equiv. 42.9 mmol) and tri-o-tolylphosphine (1.31 g, 0.2 equiv., 4.29 mmol) and K.sub.2CO.sub.3 (8.9 g, 3.0 equiv., 64.4 mmol) followed by Pd(OAc).sub.2 (0.48 g, 0.1 equiv., 2.14 mmol) at RT. The reaction mixture was stirred at 100? C. for 16 h then was filtered through celite. The resulting filtrate was diluted with water and extracted with EtOAc. The organic layers were combined and washed with ice cold water, brine, then dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The resulting residue was purified by flash chromatography (8-10% EtOAc in hexane) to afford 126 (5.7 g, 16.5 mmol, 77% yield).

[0751] LCMS m/z=246.3 [M+H-100].sup.+

[0752] .sup.1H NMR (400 MHz, DMSO-d.sub.6): ? 7.63-7.51 (m, 2H), 7.50 (d, J=1.6 Hz, 1H), 7.19 (d, J=8.0 Hz, 1H), 6.64 (m, 1H), 5.10 (br s, 1H), 4.20 (q, J=7.2 Hz, 2H), 4.05-4.03 (m, 1H), 3.13-3.12 (m, 1H), 2.78 (t, J=2.4 Hz, 2H), 1.44-1.28 (m, 12H), 1.25 (t, J=4.8 Hz, 3H).

Example 127tert-butyl 7-(3-ethoxy-3-oxopropyl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0753] ##STR00339##

[0754] To a stirred solution of compound 126 (5.7 g, 1.0 equiv., 16.5 mmol) in EtOH (50 ml) at 0? C. was added NiCl.sub.2.Math.6H.sub.2O (1.96 g, 0.5 equiv., 8.25 mmol) and NaBH.sub.4 (1.87 g, 3.0 equiv., 49.5 mmol). The reaction mixture was stirred at RT for 2 hr then concentrated in vacuo to remove the EtOH. The resulting residue was quenched with ice cold water and extracted with EtOAc. The organic layers were washed with brine, dried (Na.sub.2SO.sub.4) and concentrated in vacuo to afford 127 (5 g, 14.39 mmol, 87% yield)).

[0755] LCMS m/z: 248.3 (M+H-100)

[0756] .sup.1H NMR (400 MHz, DMSO-d.sub.6): 7.06-6.99 (m, 3H), 5.01 (br s, 1H), 4.06 (q, J=3.2 Hz, 2H), 4.03-3.97 (m, 1H), 3.13-3.12 (m, 1H), 2.78 (t, J=5.6 Hz, 2H), 2.73 (t, J=5.6 Hz, 2H), 2.68 (t, J=2.0 Hz, 2H), 1.43-1.24 (m, 12H), 1.18 (t, J=7.2 Hz, 3H),

Example 1283-(2-(tert-butoxycarbonyl)-1-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl)propanoic acid

[0757] ##STR00340##

[0758] Prepared following general procedure 7. Obtained 3.5 g, m/z=318.2 [M?H].sup.?; .sup.1H NMR (400 MHz, DMSO-d.sub.6): 12.10 (br s, 1H), 7.06-6.99 (m, 3H), 5.05 (br s, 1H), 4.04 (t, J=7.2 Hz, 1H), 3.13-3.12 (m, 1H), 2.78 (t, J=5.6 Hz, 2H), 2.73 (t, J=5.6 Hz, 2H), 2.68 (t, J=2.0 Hz, 2H), 1.43-1.24 (m, 12H),

Example 129tert-butyl 7-(3-(methoxy(methyl)amino)-3-oxopropyl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0759] ##STR00341##

[0760] Prepared following general procedure 31. Flash chromatography (0-50% EtOAc in n-hexane) obtained 3.4 g, 86% yield, m/z 263.2 [M+H-100].sup.+; .sup.1H NMR (400 MHz, DMSO-ds) 7.06-7.02 (m, 3H), 5.03 (br s, 1H), 4.04 (t, J=7.2 Hz, 1H), 3.61 (s, 3H), 3.13-3.12 (m, 1H), 3.08 (s, 3H), 2.79-2.70 (m, 4H), 2.68 (t, J=2.0 Hz, 2H), 1.43 (s, 9H), 1.36 (d, J=4.8 Hz, 3H).

Example 1301-methyl-7-(3-oxopropyl)-3,4-dihydroisoquinoline-2 (1H)-carboxylate

[0761] ##STR00342##

[0762] Prepared following general procedure 32. Crude material obtained 0.78 g, 99% yield, m/z 204.2 [M+H-100].sup.+

Example 131tert-butyl (E)-5-(3-ethoxy-3-oxoprop-1-en-1-yl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0763] ##STR00343##

[0764] To a nitrogen degassed stirred solution of compound 112 (5.0 g, 1.0 equiv., 15.33 mmol) in DMF (50 ml) was added ethyl acrylate (3.07 g, 2.0 equiv., 30.7 mmol) and tri-o-tolylphosphine (0.933 g, 0.2 equiv., 3.07 mmol) and K.sub.2CO.sub.3 (6.35 g, 3.0 equiv., 46.0 mmol) followed by Pd(OAc).sub.2 (0.344 g, 0.1 equiv., 1.533 mmol). The reaction mixture was stirred at 100? C. for 16 hr then was filtered through celite. The filtrate was diluted with water and extracted with EtOAc. The combined organic layers were washed with ice cold water, brine, dried (Na.sub.2SO.sub.4), concentrated in vacuo. The resulting residue was purified by flash chromatography (20-30% in EtOAc in n-hexane) to get 131 (3.85 g, 11.15 mmol, 72.7% yield).

[0765] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.85 (d, J=15.8 Hz, 1H), 7.61 (d, J=6.8 Hz, 1H), 7.18-7.37 (m, 2H), 6.49 (d, J=15.8 Hz, 1H), 4.97-5.21 (m, 1H), 4.20 (q, J=7.1 Hz, 2H), 3.93-4.11 (m, 1H), 3.06-3.27 (m, 1H), 2.75-2.94 (m, 2H), 1.34-1.51 (m, 12H), 1.27 (t, J=7.1 Hz, 3H).

Example 132tert-butyl 5-(3-ethoxy-3-oxopropyl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0766] ##STR00344##

[0767] To a stirred solution of 131 (2 g, 1.0 equiv., 5.79 mmol) in EtOH (25 ml) at 0? C. was added NiCl.sub.2.Math.6H.sub.2O (0.69 g, 0.5 equiv., 2.89 mmol) and NaBH.sub.4 (0.66 g, 3.0 equiv., 17.37 mmol). The reaction mixture was stirred at RT for 2 hr then concentrated in vacuo to remove the EtOH. The resulting residue was quenched with ice cold water and extracted with EtOAc. The organic layers were washed with brine, dried (Na.sub.2SO.sub.4) and concentrated in vacuo to afford 132 (1.7 g, 4.89 mmol, 85% yield).

[0768] LCMS m/z 248.3 (M+H-100)

[0769] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 6.96-7.18 (m, 3H), 4.89-5.20 (m, 1H), 3.87-4.15 (m, 3H), 3.06-3.28 (m, 1H), 2.65-2.89 (m, 4H), 2.56 (br d, J=3.3 Hz, 2H), 1.31-1.50 (m, 12H), 1.17 (t, J=7.1 Hz, 3H).

Example 1333-(2-(tert-butoxycarbonyl)-1-methyl-1,2,3,4-tetrahydroisoquinolin-5-yl)propanoic acid

[0770] ##STR00345##

[0771] Prepared following general procedure 7. Obtained 1.0 g, 68% yield, m/z=318.2 [M?H].sup.?;

[0772] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 11.91-12.32 (m, 1H), 6.98-7.20 (m, 3H), 5.05 (br d, J=5.0 Hz, 1H), 3.88-4.17 (m, 1H), 3.23 (br s, 1H), 2.65-2.84 (m, 6H), 1.37-1.49 (m, 12H).

Example 134tert-butyl 5-(3-(methoxy(methyl)amino)-3-oxopropyl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0773] ##STR00346##

[0774] Prepared following general procedure 31. Flash chromatography (0-10% MeOH in DCM) obtained 1.0 g, 88% yield, m/z 263.2 [M+H-100].sup.+

Example 135tert-butyl 1-methyl-5-(3-oxopropyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0775] ##STR00347##

[0776] Prepared following general procedure 32. Crude material obtained 0.34 g, 94% yield, m/z 204.2 [M+H-100].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.73 (t, J=1.3 Hz, 1H), 6.98-7.22 (m, 3H), 4.92-5.16 (m, 1H), 3.85-4.00 (m, 1H), 2.64-2.87 (m, 4H), 2.30-2.44 (m, 1H), 1.63-1.79 (m, 1H), 1.37-1.46 (m, 12H), 1.36 (br s, 1H).

Example 136tert-butyl 5-(3-(4-(5-fluoro-4-(5-fluoro-2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)piperidin-1-yl)propyl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0777] ##STR00348##

[0778] Prepared following general procedure 26. Obtained 370 mg, 76% yield, LCMS m/z=631 [M+H-100].

Example 1375-(3-(4-(5-fluoro-4-(5-fluoro-2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)piperidin-1-yl)propyl)-1-methyl-1,2,3,4-tetrahydroisoquinoline

[0779] ##STR00349##

[0780] Prepared following general procedure 8. Obtained 310 mg, 86% yield, LCMS m/z=531.3 [M+H].sup.+.

Example 1383-(5-(3-(4-(5-fluoro-4-(5-fluoro-2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)piperidin-1-yl)propyl)-1-methyl-3,4-dihydroisoquinolin-2(1H)-yl)-3-oxopropanenitrile

[0781] ##STR00350##

[0782] Prepared following general procedure 9. Obtained 330 mg, 65% yield, LCMS m/z=598.2 [M+H].sup.+.

Example A53(Z/E)-2-(5-(3-(4-(5-fluoro-4-(5-fluoro-2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)piperidin-1-yl)propyl)-1-methyl-1,2,3,4-tetrahydroisoquinoline-2-carbonyl)-3-(thiazol-2-yl)acrylonitrile

[0783] ##STR00351##

[0784] Prepared following general procedure 10. Obtained 40 mg, 13.8% yield, LCMS m/z=691.2 [M+H].sup.+.

Example 139tert-butyl 6-(3-(4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)carbamoyl)piperidin-1-yl)propyl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0785] ##STR00352##

[0786] Prepared following general procedure 26. Obtained 0.80 g, 32% yield, LCMS m/z=668 [M+H]).

Example 140N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(3-(1-methyl-1,2,3,4-tetrahydroisoquinolin-6-yl)propyl)piperidine-4-carboxamide

[0787] ##STR00353##

[0788] Prepared following general procedure 8. Obtained 0.70 g, 95% yield, LCMS m/z=568 [M+H].sup.+.

Example 141N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(3-(2-(2-cyanoacetyl)-1-methyl-1,2,3,4-tetrahydroisoquinolin-6-yl)propyl)piperidine-4-carboxamide

[0789] ##STR00354##

[0790] Prepared following general procedure 9. Obtained 0.70 g, 90% yield, LCMS m/z=635.6 [M+H].sup.+.

Example A54(E/Z)N-(5-(((5-(N-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(3-(2-(2-cyano-4-methyl-4-morpholinopent-2-enoyl)-1-methyl-1,2,3,4-tetrahydroisoquinolin-6-yl)propyl)piperidine-4-carboxamide

[0791] ##STR00355##

[0792] Prepared following general procedure 10. Obtained 118 mg, 27.5% yield, LCMS m/z=775 [M+H].sup.+ 1H NMR (400 MHz, DMSO-d.sub.6) 12.08-12.36 (m, 1H), 7.38 (s, 1H), 7.11-7.23 (m, 1H), 6.98-7.10 (m, 2H), 6.77-6.88 (m, 1H), 6.72 (s, 1H), 5.25-5.39 (m, 1H), 4.05 (s, 2H), 3.73-3.90 (m, 1H), 3.64 (br d, J=4.0 Hz, 4H), 3.44-3.61 (m, 2H), 2.72-3.01 (m, 4H), 2.27 (br t, J=7.0 Hz, 4H), 1.53-1.93 (m, 12H), 1.42 (br d, J=6.5 Hz, 3H), 1.09-1.33 (m, 15H).

Example A55(E/Z)N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(3-(2-(2-cyano-4,4-dimethyl-5-morpholinopent-2-enoyl)-1-methyl-1,2,3,4-tetrahydroisoquinolin-6-yl)propyl)piperidine-4-carboxamide

[0793] ##STR00356##

[0794] Prepared following general procedure 10. Obtained 18 mg, 4.7% yield, LCMS m/z=788 [M+H].sup.+

Example 142tert-butyl 6-(3-hydroxyprop-1-yn-1-yl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0795] ##STR00357##

[0796] To a mixture of compound 121 (2.2 g, 1.0 eq, 5.29 mmol) in THF (10 ml) was added TBAF (26.5 ml, 5.0 eq, 26.5 mmol) and the reaction mixture was stirred at RT for 3 h. The reaction mixture was diluted with water, and extracted with EtOAc. The combined organic layers were dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude product was purified by flash chromatography (15% to 25% EtOAc in n-hexane) to give the desired product 142 (0.98 g, 3.25 mmol, 61.4% yield) as a clear oil.

[0797] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.23-7.21 (m, 3H), 5.30 (t, J=6.00 Hz, 1H), 5.08 (m, 1H), 4.29 (d, J=6.00 Hz, 2H), 4.03-3.92 (m, 1H), 3.20-3.08 (m, 2H), 2.76-2.73 (m, 2H), 1.43-1.36 (m, 12H).

Example 143tert-butyl 6-(3-bromoprop-1-yn-1-yl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0798] ##STR00358##

[0799] To a stirred solution of compound 142 (0.2 g, 1 eq, 0.664 mmol) in DCM (10 ml) at RT under nitrogen was added triphenylphosphine (0.226 g, 1.3 eq, 0.863 mmol) and CBr.sub.4 (0.286 g, 1.3 eq, 0.863 mmol). The reaction mixture was stirred for 3 h at RT then was purified by flash column purification (7% to 10% EtOAc in n-hexane) to afford 143 (0.24 g, 0.659 mmol, 99% yield) as clear oil.

[0800] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.26 (s, 3H), 5.08 (m, 1H), 4.51 (s, 2H), 4.06-3.92 (m, 1H), 3.21-3.12 (m, 1H), 2.77-2.74 (m, 2H), 1.43-1.36 (m, 12H),

Example 144: tert-butyl 6-(3-(4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)carbamoyl)piperidin-1-yl)prop-1-yn-1-yl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0801] ##STR00359##

[0802] Prepared following general procedure 30. Obtained 190 mg, 59.7% yield, m/z=664.8 [M+H].sup.+

Example 145: N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(3-(1-methyl-1,2,3,4-tetrahydroisoquinolin-6-yl)prop-2-yn-1-yl)piperidine-4-carboxamide

[0803] ##STR00360##

[0804] Prepared following general procedure 8. Obtained 150 mg, 98% yield, m/z=564.8 [M+H].sup.+

Example 146: N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(3-(2-(2-cyanoacetyl)-1-methyl-1,2,3,4-tetrahydroisoquinolin-6-yl)prop-2-yn-1-yl)piperidine-4-carboxamide

[0805] ##STR00361##

[0806] Prepared following general procedure 9. Obtained 155 mg, 84% yield, m/z=631.4 [M+H].sup.+

Example A56: (E)-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(3-(2-(2-cyano-3-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)acryloyl)-1-methyl-1,2,3,4-tetrahydroisoquinolin-6-yl)prop-2-yn-1-yl)piperidine-4-carboxamide

[0807] ##STR00362##

[0808] Prepared following general procedure 10 in EtOH. Obtained via reverse phase preparative HPLC 6 mg, 9% yield, LCMS m/z=780.4 [M+H].sup.+

Example 147tert-butyl ((4-bromophenyl)(cyclopropyl)methyl)(methyl)carbamate

[0809] ##STR00363##

[0810] To a stirred solution of (4-bromophenyl)(cyclopropyl)methanone (15.0 g, 1 equiv., 66.6 mmol) and methylamine (2M in THF) (100 ml, 3 equiv., 200 mmol) was added titanium(IV) isopropoxide (26.3 ml, 1.3 equiv., 87 mmol) at RT. The reaction mixture was stirred at RT for 20 h, before the addition of NaBH.sub.4 (3.78 g, 1.5 equiv., 100 mmol) at 0? C. The reaction mixture was warmed to RT and stirred for 16 h before being quenched with aqueous NaHCO.sub.3 solution at 0? C. The reaction mixture was diluted with EtOAc and filtered through celite. The filtrate was washed with brine, dried (Na.sub.2SO.sub.4) and concentrated in vacuo to give 1-(4-bromophenyl)-1-cyclopropyl-N-methylmethanamine (15 g, 62.5 mmol) as a colourless oil. The crude oil was dissolved in DCM (150 ml) and triethylamine (26.1 ml, 3.0 equiv., 187 mmol) was added at 0? C. To this mixture Boc-anhydride (21.5 ml, 1.5 equiv., 94 mmol) was added slowly and the resulting reaction mixture was stirred for 24 h at RT under N.sub.2 atmosphere. The reaction mixture was diluted with DCM and extracted with water. The organic layer was dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude product was purified by flash chromatography (0-100% EtOAc in n-hexane) to give desired product 147 (16.5 g, 48.5 mmol, 78% yield).

[0811] LCMS: Mass found; [M+H].sup.+ 340 and [M+H+2].sup.+ 342.

Example 148tert-butyl (cyclopropyl(4-vinylphenyl)methyl)(methyl)carbamate

[0812] ##STR00364##

[0813] To a stirred solution of compound 147 (16.0 g, 1 equiv., 47 mmol), potassium vinyltrifluoroborate (18.9 g, 3.0 equiv., 141 mmol) and aqueous Cs.sub.2CO.sub.3 (2 M in water) (23.51 ml, 1 equiv., 47 mmol) in 1,4-dioxane (180 ml) was purged with nitrogen for 20 min followed by the addition of PdCl.sub.2(dpp)-DCM adduct (3.84 g, 0.1 equiv., 4.70 mmol). The reaction mixture was stirred at 85? C. for 18 h then was quenched with water and extracted with EtOAc. The combined organic layers were washed with brine, dried (Na.sub.2SO.sub.4) and concentrated in vacuo to give 148 (11.0 g, 38.3 mmol, 81% yield).

[0814] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.70-7.46 (m, 2H), 7.36-7.32 (m, 2H), 6.73 (dd, J=17.8, 10.9 Hz, 1H), 5.82 (dd, J=17.8, 0.8 Hz, 1H), 5.25 (dd, J=10.9, 0.8 Hz, 1H), 4.23-4.12 (m, 1H), 3.28-3.18 (m, 1H), 2.68 (s, 3H), 1.39 (br s, 9H), 0.79-0.75 (m, 2H), 0.41-0.31 (m, 2H).

Example 149tert-butyl (cyclopropyl(4-formylphenyl)methyl)(methyl)carbamate

[0815] ##STR00365##

[0816] To a solution of compound 148 (11.0 g, 1.0 equiv., 38.3 mmol) in 1,4-dioxane (300 ml) and water (30 ml) was added sodium periodate (16.4 g, 2.0 equiv., 77 mmol) and N-methylmorpholine (1.94 g, 0.5 equiv., 19.14 mmol) at 0? C. followed by slow addition of osmium tetroxide (30.0 ml, 0.1 equiv., 3.83 mmol). The reaction mixture was and stirred at RT for 16 h then concentrated in vacuo. The crude residue was diluted with water:EtOAc (1:1), filtered and extracted with EtOAc. The combined organic layers were dried (Na.sub.2SO.sub.4), concentrated in vacuo then purified by flash columnatography (0-25% EtOAc in n-hexane). The appropriate fractions were combined and concentrated in vacuo to give 149 (8.0 g, 27.6 mmol, 72.2% yield).

Example 150tert-butyl (cyclopropyl(4-((4-hydroxypiperidin-1-yl)methyl)phenyl)methyl)(methyl) carbamate

[0817] ##STR00366##

[0818] Prepared following general procedure 26. Obtained 3.0 g, 55% yield, m/z=375.4 [M+H].sup.+

Example 151tert-butyl (cyclopropyl(4-((4-oxopiperidin-1-yl)methyl)phenyl)methyl)(methyl) carbamate

[0819] ##STR00367##

[0820] To a stirred solution of oxalyl chloride (0.11 ml, 1.2 equiv., 1.28 mmol) in DCM (10 ml) at ?78? C. was added DMSO (0.19 ml, 2.1 equiv., 2.67 mmol) and the reaction was stirred for 10 min. To this mixture was added compound 150 (0.40 g, 1.0 equiv., 1.07 mmol) in DCM (10 ml) and the mixture was stirred for 30 min at ?78? C. The reaction mixture was quenched with Et.sub.3N (0.744 ml, 5.0 equiv., 5.34 mmol) and allowed to warm to RT. Saturated aqueous ammonium chloride solution was added then the organic phase was washed with water, brine, dried (Na.sub.2SO.sub.4), and concentrated in vacuo. The crude residue was purified by flash chromatography (0-10% MeOH in DCM). The desired fractions were combined and concentrated in vacuo to give 151 (0.2 g, 0.53 mmol, 49.8% yield).

[0821] LCMS m/z=[M+H].sup.+ 373.2

Example 152tert-butyl ((4-((4-(((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)methyl)-[1,4-bipiperidin]-1-yl)methyl)phenyl)(cyclopropyl)methyl)(methyl) carbamate

[0822] ##STR00368##

[0823] Prepared following general procedure 26. Obtained 180 mg, 91% yield, m/z=723.6 [M+H].sup.+, .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.89 (br s, 1H), 8.05 (s, 1H), 7.31 (m, 4H), 6.91 (s, 1H), 6.73 (s, 1H), 4.13-4.04 (m, 1H), 3.51 (m, 2H), 3.33 (m, 2H), 3.18-2.92 (m, 6H), 2.68-2.51 (m, 4H), 1.92-1.82 (m, 10H), 1.39 (s, 9H), 1.23-1.16 (m, 12H), 0.90-0.71 (m, 4H).

Example 1535-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-N-((1-(4-(cyclopropyl(methylamino)methyl) benzyl)-[1,4-bipiperidin]-4-yl) methyl)thiazol-2-amine

[0824] ##STR00369##

[0825] Prepared following general procedure 8. Obtained 130 mg, 89% yield, m/z=623.6 [M+H].sup.+

Example 154N-((4-((4-(((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)methyl)-[1,4-bipiperidin]-1-yl)methyl)phenyl)(cyclopropyl)methyl)-2-cyano-N-methylacetamide

[0826] ##STR00370##

[0827] Prepared following general procedure 9. Obtained 120 mg, 79% yield, m/z=690.4 [M+H].sup.+, .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.12 (br s, 1H), 7.19-7.50 (m, 4H), 6.91 (s, 1H), 6.73 (s, 1H), 4.74 (br d, J=10.1 Hz, 1H), 4.04-4.27 (m, 3H), 3.94 (s, 2H), 3.51 (br s, 2H), 3.02-3.22 (m, 3H), 2.64-2.99 (m, 6H), 1.61-2.09 (m, 7H), 1.33-1.57 (m, 4H), 1.14-1.33 (m, 9H), 0.72-0.96 (m, 4H), 0.22-0.65 (m, 3H).

Example A57(E/Z)N-((4-((4-(((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)methyl)-[1,4-bipiperidin]-1-yl)methyl)phenyl)(cyclopropyl)methyl)-2-cyano-N-methyl-3-phenylacrylamide

[0828] ##STR00371##

[0829] Prepared following general procedure 10 in THF. Obtained via reverse phase preparative HPLC, 11 mg, 16% yield, LCMS m/z=778.5 [M+H]; .sup.1H NMR (400 MHz, DMSO-d.sub.6) 8.21 (s, 1H), 7.93 (br d, J=5.3 Hz, 2H), 7.56 (br s, 2H), 7.47 (br d, J=7.0 Hz, 1H), 7.40 (br s, 2H), 7.27-7.37 (m, 3H), 6.89 (s, 1H), 6.72 (s, 1H), 3.93 (s, 2H), 3.43 (br s, 4H), 2.93-3.08 (m, 4H), 2.85 (br s, 4H), 2.09 (br t, J=10.9 Hz, 2H), 1.90 (br t, J=10.9 Hz, 2H), 1.60-1.71 (m, 4H), 1.38-1.56 (m, 5H), 1.22 (s, 9H), 1.06-1.17 (m, 2H), 0.38-0.90 (m, 4H).

Example 155tert-butyl ((4-((4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl) carbamoyl)-[1,4-bipiperidin]-1-yl)methyl)phenyl)(cyclopropyl)methyl) (methyl)carbamate

[0830] ##STR00372##

[0831] Prepared following general procedure 26. Obtained 180 mg, 91% yield, m/z=737.6 [M+H].sup.+

Example 156N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(4-(cyclopropyl(methylamino)methyl)benzyl)-[1,4-bipiperidine]-4-carboxamide hydrochloride

[0832] ##STR00373##

[0833] Prepared following general procedure 8. Obtained 140 mg, 79% yield, m/z=637.6 [M+H].sup.+

Example 157N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(4-((2-cyano-N-methylacetamido)(cyclopropyl)methyl)benzyl)-[1,4-bipiperidine]-4-carboxamide

[0834] ##STR00374##

[0835] Prepared following general procedure 9. Obtained 120 mg, 62% yield, m/z=704.2 [M+H].sup.+

Example A58: (E/Z)N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(4-((2-cyano-N-methyl-3-phenylacrylamido)(cyclopropyl)methyl)benzyl)-[1,4-bipiperidine]-4-carboxamide

[0836] ##STR00375##

[0837] Prepared following general procedure 10 in EtOH. Obtained via reverse phase preparative HPLC 9.6 mg, 14% yield, LCMS m/z=792.5 [M+H]; .sup.1H NMR (400 MHz, DMSO-d.sub.6) 12.13-12.31 (m, 1H), 7.88-7.98 (m, 2H), 7.78-7.86 (m, 1H), 7.52-7.60 (m, 3H), 7.39-7.45 (m, 2H), 7.38 (s, 1H), 7.30-7.36 (m, 2H), 6.72 (s, 1H), 4.05 (s, 2H), 3.44 (s, 4H), 2.94-3.03 (m, 2H), 2.87 (br dd, J=18.9, 11.1 Hz, 4H), 2.06-2.28 (m, 4H), 1.91 (br t, J=10.5 Hz, 2H), 1.65-1.80 (m, 4H), 1.50-1.62 (m, 3H), 1.39-1.49 (m, 2H), 1.18 (s, 9H), 0.80-0.90 (m, 1H), 0.36-0.71 (m, 3H).

Example 158tert-butyl 7-(3-(4-(5-fluoro-4-(5-fluoro-2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)piperidin-1-yl)propyl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0838] ##STR00376##

[0839] Prepared following general procedure 26 (using amine as prepared in WO2014139328, the entire contents of which are incorporated herein by reference). Obtained 500 mg, 63.5% yield, m/z=631.3 [M+H-100].sup.+

Example 1597-(3-(4-(5-fluoro-4-(5-fluoro-2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)piperidin-1-yl) propyl)-1-methyl-1,2,3,4-tetrahydroisoquinoline

[0840] ##STR00377##

[0841] Prepared following general procedure 8. Obtained 500 mg, 81% yield, m/z=531.3 [M+H].sup.+

Example 1603-(7-(3-(4-(5-fluoro-4-(5-fluoro-2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)piperidin-1-yl)propyl)-1-methyl-3,4-dihydroisoquinolin-2(1H)-yl)-3-oxopropanenitrile

[0842] ##STR00378##

[0843] Prepared following general procedure 10. Obtained 370 mg, 52% yield, m/z=598.2 [M+H].sup.+

Example A59(E/Z)-2-(7-(3-(4-(5-fluoro-4-(5-fluoro-2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)piperidin-1-yl)propyl)-1-methyl-1,2,3,4-tetrahydroisoquinoline-2-carbonyl)-3-(thiazol-2-yl)acrylonitrile

[0844] ##STR00379##

[0845] Prepared following general procedure 10 in EtOH. Obtained via reverse phase preparative HPLC 47 mg, 11% yield, LCMS m/z=693.2 [M+H].

Example 161tert-butyl (1-(4-(3-(4-(5-fluoro-4-(5-fluoro-2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-2-yl) piperidin-1-yl)propyl)phenyl)butyl)(methyl)carbamate

[0846] ##STR00380##

[0847] Prepared following general procedure 26 (using amine as prepared in WO2014139328, the entire contents of which are incorporated herein by reference). Obtained 1.4 g, 53.4% yield, m/z=647.4 [M+H].sup.+

Example 1621-(4-(3-(4-(5-fluoro-4-(5-fluoro-2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)piperidin-1-yl)propyl)phenyl)-N-methylbutan-1-amine

[0848] ##STR00381##

[0849] Prepared following general procedure 8. Obtained 0.91 g, 98% yield, m/z=547.4[M+H].sup.+

Example 1632-cyano-N-(1-(4-(3-(4-(5-fluoro-4-(5-fluoro-2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-2-yl) piperidin-1-yl)propyl)phenyl)butyl)-N-methylacetamide

[0850] ##STR00382##

[0851] Prepared following general procedure 10. Obtained 850 mg, 35.5% yield, m/z=614.4 [M+H].sup.+

Example A60(E/Z)-2-cyano-N-(1-(4-(3-(4-(5-fluoro-4-(5-fluoro-2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)piperidin-1-yl)propyl)phenyl)butyl)-N-methyl-3-(thiophen-2-yl)acrylamide

[0852] ##STR00383##

[0853] Prepared following general procedure 10 in EtOH. Obtained via reverse phase preparative HPLC, 45 mg, 13% yield, LCMS m/z=708.2 [M+H].

Example 164: [5-[4-[2-fluoro-5-[(4-oxo-3H-phthalazin-1-yl)methyl]benzoyl]piperazin-1-yl]-5-oxo-pentyl]ammonium;chloride

[0854] ##STR00384##

[0855] 4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazin-1(2H)-one hydrochloride salt (1.00 eq, 61 mg, 0.153 mmol) and 5-(Boc-amino)valeric acid (1.00 eq, 33 mg, 0.153 mmol) were suspended n DMF (0.6 mL). HATU (1.00 eq, 58 mg, 0.153 mmol) was added, followed by N,N-Diisopropylethylamine (5.00 eq, 0.13 mL, 0.765 mmol). The reaction mixture was left to react for 1 hour, diluted to 2 mL with ACN/H.sub.2O and purified by preparative HPLC using a gradient from 10% to 95% of ACN in Water with 0.1% of formic acid over 10 minutes. Fractions contained the desired product were evaporated to dryness. The residue was dissolved in DCM (2 mL) and treated with HCl in Dioxane (52.3 eq, 2.0 mL, 8.00 mmol) 4 M for 1 hour. Volatiles were removed to obtain the title compound (m/z=466.22, (M+H.sup.+), 77 mg, 56.5% yield.

Additional Examples

[0856]

TABLE-US-00018 Example m/z number Structure/Preparation (M + H).sup.+ 165 [00385] embedded image 494.3 166 [00386] 522.2

Example A61 (E)-2-cyano-N-(1-(4-(3-((5-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-5-oxopentyl)amino)-3-oxopropyl)phenyl)butyl)-N-methyl-3-(thiazol-2-yl)acrylamide

[0857] ##STR00387##

[0858] To a solution of compound 164 (0.05 M in DMF, 0.50 mL, 0.0252 mmol), compound 44 (1.00 eq, 10 mg, 0.0252 mmol) and N,N-Diisopropylethylamine (5.00 eq, 0.022 mL, 0.126 mmol) in DMF (0.5 mL) were added, followed by HATU (1.00 eq, 9.6 mg, 0.0252 mmol). The reaction was left to stir at r.t. for 2 hours, then diluted with MeOH and purified by preparative HPLC using a gradient from 5% to 95% of ACN in Water containing 0.1% of formic acid to obtain the desired product (m/z=845.35, (M+H.sup.+), as white solid, 14.28 mg, 65% yield.

Additional Examples

[0859]

TABLE-US-00019 Example m/z number Structure/Preparation (M + H).sup.+ A62 [00388] embedded image 873.4 A63 [00389] 901.4

Example 167: tert-butyl (S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-((2-(trimethylsilyl)ethoxy)carbonyl)pyrrolidin-2-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0860] ##STR00390##

[0861] To a solution of 2-trimethylsilylethyl (2S)-2-(hydroxymethyl)pyrrolidine-1-carboxylate (1.69 g, 2 equiv., 6.88 mmol) in THF (10 mL) was added sodium t-butoxide (661 mg, 2 equiv., 6.88 mmol) followed by a portion-wise addition of tert-butyl (2S)-4-[7-(8-chloro-1-naphthyl)-2-methylsulfinyl-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate (2 g, 1 equiv., 3.44 mmol, prepared as described in WO2019/99524, the entire contents of which are incorporated herein by reference). The reaction mixture was stirred at rt for 16 h then was quenched with water and extracted with EtOAc. The combined organics were washed with brine, dried over MgSO.sub.4, filtered and concentrated in vacuo. Purification by flash chromatography (DCM/MeOH (0-20%)) yielded the desired product as a pale brown solid (2.11 g, 2.77 mmol, 80.4% yield).

[0862] m/z=762.8 (M+H).sup.+

Example 168: tert-butyl (S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-pyrrolidin-2-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0863] ##STR00391##

[0864] To a solution of compound 167 (2.11 g, 1 equiv., 2.77 mmol) in THF (10 mL) was added TBAF (1M in THF) (3.6 mL, 1.3 equiv., 3.60 mmol) and the reaction was stirred at rt for 16 h. Additional TBAF (6.0 mL, 2.2 equiv., 6.0 mmol) added and reaction was stirred at rt for 4 h. The reaction was diluted in DCM and was washed with sat. NH.sub.4Cl. The organics were dried (MgSO.sub.4), filtered and concentrated in vacuo. Purification by flash chromatography (DCM/MeOH (0-20%)) yielded the crude product, which was dissolved in EtOAc and washed with brine, dried (MgSO.sub.4) and concentrated in vacuo to give the desired product (0.90 g, 1.02 mmol, 36.8% yield) as a brown solid.

[0865] A sample of the crude was purified by preparative HPLC using a gradient from 5% to 95% of Acetonitrile in water (containing 0.1% of formic acid) for analytical purposes.

[0866] NMR: .sup.1H NMR (400 MHz, CDCl3) ? 7.77-7.74 (m, 1H), 7.64-7.58 (m, 1H), 7.53-7.49 (m, 1H), 7.47-7.37 (m, 1H), 7.35-7.31 (m, 1H), 7.25-7.16 (m, 1H), 4.64-4.52 (m, 2H), 4.50-4.33 (m, 1H), 4.32-4.18 (m, 1H), 4.14-3.96 (m, 2H), 3.94-3.70 (m, 2H), 3.60-3.50 (m, 1H), 3.44-3.32 (m, 2H), 3.29-3.17 (m, 2H), 3.16-3.01 (m, 4H), 2.96-2.86 (m, 1H), 2.75-2.65 (m, 2H), 2.62-2.50 (m, 1H), 2.20-1.85 (m, 4H), 1.52-1.48 (br, 9H).

[0867] m/z=618.7 (M+H).sup.+

Example 169: 2-(trimethylsilyl)ethyl (4-hydroxybutyl)carbamate

[0868] ##STR00392##

[0869] To a solution of 4-amino-1-butanol (1.0 mL, 1 equiv., 11.2 mmol) and triethylamine (2.0 mL, 1.3 equiv., 14.6 mmol) in DCM (40 mL) was added (2,5-dioxopyrrolidin-1-yl) 3-trimethylsilylpropanoate (3.0 g, 1.1 equiv., 12.3 mmol) portion-wise. The reaction was quenched with sat. NH.sub.4Cl and diluted with DCM. The layers were separated, and the organics dried and concentrated in vacuo. Purification by flash chromatography (Hex/EtOAc (0-100%)) yielded the desired product as a colourless oil (2.5 g, 10.8 mmol, 96.3% yield).

[0870] .sup.1H NMR (400 MHz, CDCl3) ? 4.20-4.09 (m, 2H), 3.72-3.61 (m, 2H), 3.24-3.17 (m, 2H), 1.66-1.52 (m, 4H), 1.03-0.93 (m, 2H), 0.03 (s, 9H).

Example 170: 2-(trimethylsilyl)ethyl (4-oxobutyl)carbamate

[0871] ##STR00393##

[0872] To a solution of dimethyl sulfoxide (0.11 ml, 2.5 eq, 1.5 mmol) in DCM (2.5 ml) was added oxalyl chloride (0.6 ml, 2 eq, 1.2 mmol) dropwise at ?78? C. The mixture was stirred for 20 min and a solution of compound 169 (140 mg, 1 eq, 0.6 mmol) in DCM (3.0 ml) was added dropwise at ?78? C. The mixture was stirred for an additional 45 min. DIPEA (0.82 ml, 8 eq, 4.8 mmol) was added dropwise and the mixture was warmed up at 0? C. and was stirred for an additional 2 h. The mixture was quenched with water (10 ml) and the aqueous phase was extracted with DCM. The solvent was removed under reduced pressure to give the product in quantitative yield.

[0873] NMR: .sup.1H NMR (400 MHz, CDCl.sub.3) ? 11.28 (s, 1H), 4.27-4.17 (m, 2H), 3.61-3.49 (m, 1H), 3.42-3.26 (m, 1H), 2.13-1.78 (m, 4H), 1.06-0.98 (m, 2H), 0.06-0.02 (m, 9H).

[0874] m/z=254.2 (M+Na).sup.+

Example 171: tert-butyl (S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-(4-(((2-(trimethylsilyl)ethoxy)carbonyl)amino)butyl)pyrrolidin-2-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0875] ##STR00394##

[0876] To a solution of compound 170 (40 mg, 1 eq, 0.065 mol) in DCM (0.5 ml) was added a solution of 2-trimethylsilylethyl N-(4-oxobutyl)carbamate (30 mg, 2.00 eq, 0.129 mmol) at room temperature. The mixture was stirred for 20 min and sodium triacetoxyborohydride (34 mg, 2.5 eq, 0.162 mmol) was added and the mixture which was stirred for an additional 2 h at room temperature.

[0877] The reaction was quenched with water and dissolved in DCM. The organic phase was washed with water and the organic phase was dried and evaporated under reduced pressure. The crude product was purified by HPLC (using a gradient from 5% to 95% of acetonitrile in water containing 0.1% of formic acid over 10 minutes) to give pure product as a yellowish solid (42 mg, 0.050 mmol, 78% yield)

[0878] NMR: .sup.1H NMR (400 MHz, CDCl.sub.3) ? 7.76-7.73 (m, 1H), 7.63-7.58 (m, 1H), 7.53-7.48 (m, 1H), 7.47-7.41 (m, 1H), 7.35-7.30 (m, 1H), 7.25-7.19 (m, 1H), 4.90-4.80 (m, 1H), 4.65-4.55 (m, 1H), 4.54-4.48 (m, 1H), 4.45-4.34 (m, 1H), 4.15-4.02 (m, 3H), 4.01-3.72 (m, 3H), 3.70-3.61 (m, 1H), 3.60-3.52 (m, 1H), 3.50-3.23 (m, 3H), 3.23-3.10 (m, 4H), 3.09-2.89 (m, 4H), 2.81-2.65 (m, 2H), 2.62-2.53 (m, 1H), 2.31-2.15 (m, 2H), 2.13-1.75 (m, 6H), 1.66-1.55 (m, 2H), 1.55-1.46 (m, 9H), 0.02-?0.02 (br, 9H).

[0879] m/z=833.9 (M+H).sup.+

Example 172: tert-butyl (2S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((2S)-1-(4-(3-(4-(1-((E)-2-cyano-N-methyl-3-(thiazol-2-yl)acrylamido)butyl)phenyl)propanamido)butyl)pyrrolidin-2-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0880] ##STR00395##

[0881] To a solution of compound 171 (50 mg, 1 equiv., 0.06 mmol) in DMF (1 mL) was added TBAF (1M in THF) (0.12 mL, 2 equiv., 0.12 mmol) and the reaction was stirred at rt for 2 h. HPLC analysis showed complete deprotection. The reaction was diluted in DCM and was washed with sat. NaHCO.sub.3. The organics were dried (MgSO.sub.4), filtered and concentrated in vacuo. The crude was dissolved in DMF (0.5 mL), compound 44 (24 mg, 0.06 mmol) was added, followed by HATU (26 mg, 1.5 equiv., 0.07 mmol) and DIPEA (31 ?L, 0.18 mmol). The reaction mixture was stirred for 30 min then was purified on preparative HPLC (using a gradient from 5% to 95% of acetonitrile in water containing 0.1% of formic acid over 10 minutes) to give the purified product 172 (55 mg, 0.05 mmol) as a pale brown solid.

[0882] m/z=1069.2 (M+H

Example 173: (E)-N-(1-(4-(3-((4-((S)-2-(((7-(8-chloronaphthalen-1-yl)-4-((S)-3-(cyanomethyl)piperazin-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)pyrrolidin-1-yl)butyl)amino)-3-oxopropyl)phenyl)butyl)-2-cyano-N-methyl-3-(thiazol-2-yl)acrylamide (hydrochloride salt)

[0883] ##STR00396##

[0884] To a solution of 172 (55 mg, 1 equiv., 0.05 mmol) in DCM (0.2 mL) was added HCl (4M in dioxane) (0.2 mL, 15.5 equiv., 0.8 mmol). The reaction was stirred at rt for 1 hr then the volatiles were removed in vacuo to give the desired product 173 (25 mg, 0.02 mmol, 46.6% yield) as a tan solid.

[0885] m/z=969.1 (M+H).sup.+

Example A64: (E)-N-[1-[4-[3-[4-[(2S)-2-[[7-(8-chloro-1-naphthyl)-4-[(3S)-3-(cyanomethyl)-4-prop-2-enoyl-piperazin-1-yl]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl]oxymethyl]pyrrolidin-1-yl]butylamino]-3-oxo-propyl]phenyl]butyl]-2-cyano-N-methyl-3-thiazol-2-yl-prop-2-enamide

[0886] ##STR00397##

[0887] To a solution of compound 173 (12 mg, 1 equiv., 0.01 mmol) and triethylamine (8.3 uL, 5 equiv., 0.06 mmol) in DCM (0.5 mL) was added prop-2-enoyl chloride (2.7 uL, 3 equiv., 0.04 mmol). The reaction mixture was stirred for 30 min at rt then concentrated in vacuo. Purification by preparative HPLC (using a gradient from 5% to 95% of acetonitrile in water containing 0.1% of formic acid over 10 minutes) gave the purified product A64 (1.7 mg, 0.002 mmol, 13.3% yield) as a white solid

[0888] m/z=1023.2 (M+H).sup.+

Example 174tert-butyl (1-(4-(3-(4-(2,6-dimethoxy-4-(2-methyl-1-oxo-1,2-dihydro-2,7-naphthyridin-4-yl)benzyl)piperazin-1-yl)propyl)phenyl)butyl)(methyl)carbamate

[0889] ##STR00398##

[0890] Prepared following general procedure 26 using amine synthesised according to J. Med. Chem. 2019, 62, 2, 699-726. Obtained 72 mg, 66% yield, LCMS m/z=698.9 [M+H].sup.+.

Example 1754-(3,5-dimethoxy-4-((4-(3-(4-(1-(methylamino)butyl)phenyl)propyl)piperazin-1-yl)methyl)phenyl)-2-methyl-2,7-naphthyridin-1(2H)-one

[0891] ##STR00399##

[0892] Prepared following general procedure 8. Used without further purification. LCMS m/z=598.7 [M+H].sup.+.

Example 176: 2-cyano-N-(1-(4-(3-(4-(2,6-dimethoxy-4-(2-methyl-1-oxo-1,2-dihydro-2,7-naphthyridin-4-yl)benzyl)piperazin-1-yl)propyl)phenyl)butyl)-N-methylacetamide

[0893] ##STR00400##

[0894] Prepared following general procedure 9. Obtained 12 mg, 17% yield (over 2 steps).

[0895] LCMS m/z=665.8 (M+H).sup.+.

Example A65: (E)-2-cyano-N-(1-(4-(3-(4-(2,6-dimethoxy-4-(2-methyl-1-oxo-1,2-dihydro-2,7-naphthyridin-4-yl)benzyl)piperazin-1-yl)propyl)phenyl)butyl)-N-methyl-3-(thiazol-2-yl)acrylamide

[0896] ##STR00401##

[0897] Prepared following general procedure 10. Obtained 7 mg, 50% yield. LCMS m/z=760.9 [M+H].sup.+.

Example A66(E)-2-cyano-N-(1-(4-(3-(4-(2,6-dimethoxy-4-(2-methyl-1-oxo-1,2-dihydro-2,7-naphthyridin-4-yl)benzyl)piperazin-1-yl)-3-oxopropyl)phenyl)butyl)-N-methyl-3-(thiazol-2-yl)acrylamide

[0898] ##STR00402##

[0899] Prepared following general procedure 31 using amine synthesised according to J. Med. Chem. 2019, 62, 2, 699-726. Obtained 12 mg, 45% yield, LCMS m/z 774.8 [M+H].sup.+.

Example 1771-isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-pyrazolo[4,3-c]pyridine-3-carboxylate

[0900] ##STR00403##

[0901] To a stirred solution of methyl 6-chloro-1-isopropyl-1H-pyrazolo[4,3-c]pyridine-3-carboxylate (prepared according to WO2014210354, the entire contents of which are incorporated herein by reference) (3 g, 1.0 equiv, 11.83 mmol) and 2-(4-methoxypiperidin-1-yl)pyrimidin-4-amine (prepared according to WO2014210354, the entire contents of which are incorporated herein by reference) (2.96 g, 1.2 equiv., 14.19 mmol) in 1,4-dioxane (30 ml), was added Cs.sub.2CO.sub.3 (7.71 g, 2.0 equiv., 23.65 mmol). The reaction mixture was degassed with N.sub.2 for 15 min followed by the addition of XPhos (0.56 g, 0.1 equiv., 1.18 mmol) and Pd.sub.2(dba).sub.3 (1.62 g, 0.15 equiv., 1.77 mmol) at RT. The reaction mixture was heated to 100? C. for 16 h then was filtered through celite and the solid was washed with EtOAc. The filtrate was concentrated in vacuo and the resulting residue was purified by flash chromatography (EtOAc in n-hexane (30% to 40%)) as an eluent. The product containing fractions were concentrated in vacuo obtain compound 177 (3.1 g, 7.01 mmol, 59.3% yield).

[0902] LCMS m/z=426.0 [M+H].sup.+.

Example 1781-isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-pyrazolo[4,3-c]pyridine-3-carboxylic acid

[0903] ##STR00404##

[0904] Prepared following general procedure 7 in THF, MeOH and H.sub.2O. Obtained 2.1 g, 71.6% yield, LCMS m/z=412.3 [M+H].

Example 179(3-(4-(3-aminopropyl)piperazin-1-yl)propyl)carbamate

[0905] ##STR00405##

[0906] Prepared following general procedure 5 in acetone. Obtained via reverse phase preparative HPLC, 800 mg, 24% yield, LCMS m/z=301.2 [M+H].sup.+.

Example 180tert-butyl (3-(4-(3-(1-isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-pyrazolo[4,3-c]pyridine-3-carboxamido) propyl) piperazin-1-yl)propyl)carbamate

[0907] ##STR00406##

[0908] Prepared following general procedure 31. Obtained 120 mg, 62.6% yield, LCMS m/z=694.7 [M+H].sup.+.

Example 181N-(3-(4-(3-aminopropyl)piperazin-1-yl)propyl)-1-isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-pyrazolo[4,3-c]pyridine-3-carboxamide hydrochloride

[0909] ##STR00407##

[0910] Prepared following general procedure 8. Obtained 100 mg, 85% yield, LCMS m/z=594.7 [M+H].sup.+.

Example A67(E)-N-(3-(4-(3-(3-(4-(1-(2-cyano-N-methyl-3-(thiazol-2-yl)acrylamido) butyl)phenyl)propanamido)propyl)piperazin-1-yl)propyl)-1-isopropyl-6-((2-(4-methoxypiperidin-1-yl)pyrimidin-4-yl)amino)-1H-pyrazolo[4,3-c]pyridine-3-carboxamide

[0911] ##STR00408##

[0912] Prepared following general procedure 31. Purified by preparative HPLC. Obtained 45 mg, 28.1% yield, LCMS m/z=973.7 [M+H].sup.+.

PART BBIOLOGICAL DATA

[0913] The bifunctional compounds were assayed to investigate their ability to degrade target proteins in accordance with the following general procedures.

[0914] 1.1 Assay 1Degradation of HiBit-BRD4 in HEK293

[0915] HEK293 containing a HiBit insertion for BRD4 were plated in 384-well tissue culture plates at a density of 8?10.sup.4 per well in a volume of 36 ?L and incubated overnight at 37? C. and 5% CO.sub.2. Wells were treated with test compounds for 6 h prior to addition of the NanoLuc substrate and reading on a ClarioSLARIOstar Plus. Degradation data was plotted and analysed using Prism 86 (Graphpad).

[0916] 1.2 Assay 2CDK9 Degradation in MV4;11

[0917] MV4;11 (0.8?10.sup.6 cells/mL) were seeded in 6-well plates (3 mL IMDM supplemented with 10% FBS and L-glutamine) overnight before treatment with compounds at the desired concentration and with a final DMSO concentration of 0.1% v/v. After 8 h incubation time, cells were washed with DPBS (Gibco) and lysed using 85 ?L RIPA buffer (Sigma-Aldrich) supplemented with cOmplete Mini EDTA-free protease inhibitor cocktail (Roche) and benzonase. Lysates were clarified by centrifugation (20000 g, 10 min, 4? C.) and the total protein content of the supernatant was quantified using a BCA assay. Samples were prepared using equal amounts of total protein and LDS sample buffer (Invitrogen). For immunoblot analysis, the following antibodies were used: anti-CDK9 (CST-2316S, 1:1,000 dilution), and anti-GAPDH hFAB-rhodamine (BioRad, 12004168, 1:5000 dilution). Band intensities were normalized to the GAPDH loading control and reported as % of the average 0.1% DMSO vehicle intensity.

[0918] 1.3 Assay 3MV4;11 Cell Viability Using CellTiter-Glo Assay

[0919] The anti-proliferative effects of representative compounds were measured using the CellTiter-Glo assay (Promega). MV4;11 cells were seeded into sterile, white, clear-bottomed 384-well cell-culture microplates (Greiner Bio-one), at 2? concentration in IMDM media and a volume of 25 ?L. Test compounds were serially diluted (11-pt dose-response from 1 ?M) in IMDM media to 2? concentration, then added to cells to make a final volume of 50 ?L. Final DMSO concentration was 0.05%. After 48 h incubation, 25 ?L of CellTiter-Glo reagent was added to each well. Following a 15 minutes incubation the luminescence signal was read on a CLARIOStar Plus. Data was processed and dose-response curves were generated using Prism 8 (Graphpad).

[0920] 1.4 Assay 4Endpoint Degradation Using HiBit-CDK9.

[0921] HiBit-CDK9 HEK293 cells were diluted in OptiMEM media with 4% FBS to 2.2?10.sup.5 cells/mL, and dispensed into a sterile, white, clear-bottomed 384-well cell-culture microplate (Greiner Bio-one) at a volume of 36 ?L. Plates were incubated for 24 h at 37? C. Test compounds were serially diluted in OptiMEM to 10? their desired final concentration and added to the assay plate at a volume of 4 ?L. After 6 h incubation, Nanoluc substrate was diluted to 1? in OptiMEM media and added to each well at a volume of 10 ?L. The plate was read immediately on a Clariostar-Plus (BMG). Dose-response curves were generated in Prism 8 (Graphpad).

[0922] 1.5 Assay 5Degradation of BRD9 by Immunoblot.

[0923] HEK293 cells (0.4?10.sup.6) were seeded in a 12-well plate (1 mL medium) overnight prior to 24 h treatment with test compounds at the desired concentration. After 24 h incubation, cells were washed with PBS and lysed with RIPA buffer (Sigma) supplemented with 1? protease inhibitor cocktail (Roche) and 1 U/mL Benzonase (Merck). Lysates were clarified by centrifugation (17,000?g, 20 min, 4? C.) and the total protein content of the supernatant was quantified using a BCA assay. Samples were prepared using equal amounts of total protein and LDS sample buffer (Invitrogen). Samples were resolved by SDS-PAGE using NuPAGE 4-12% Bis-Tris midi gels (Invitrogen, followed by transfer to Amersham Protran 0.45 NC nitrocellulose membrane (GE Healthcare) using wet transfer. Precision Plus Protein All Blue (Bio-Rad) protein ladder was used as a standard. The membrane was blocked with 5% powdered skimmed milk (Marvel) in Tris-buffered saline with 0.1% Tween-20 (TBST). Blots were probed (overnight at 4? C.) using the following primary antibodies (diluted in 5% BSA in TBST) as appropriate: anti-BRD9 (Bethyl A303-781A, 1:1000) and anti-BRD7 (Cell Signalling #15125, rabbit, 1:2000). The next day, blots were washed with TBST and incubated (1 h at RT) with anti-Tubulin hFAB-rhodamine (BioRad, 12004166. 1:30000) and and anti-GAPDH hFAB-rhodamine (#12004168, 1:5000) primary antibodies, plus either anti-rabbit IRDye 800CW (Licor 1:10,000 dilution) or anti-mouse IRDye 800CW (Licor 1:10,000 dilution) secondary antibody. Blots were visualised using a Bio-Rad ChemiDoc MP Imaging System, and band quantification was performed using Image Studio software (LiCor). Band intensities were normalized to the loading controls and reported as % of the average 0.1% DMSO vehicle intensity. Degradation data was plotted and analysed using Prism (Graphpad, version 8).

[0924] 1.6 Assay 6Degradation of KRas-G12C by Immunoblot.

[0925] MIA-PaCa-2 cells were seeded in 6- or 12-well plates overnight prior to 24 h treatment with test compounds at the desired concentration (DMSO final concentration 0.1%). After 24 h incubation, cells were washed with PBS and lysed with RIPA buffer (Sigma) supplemented with 1 mM MgCl.sub.2, 1 U/mL Benzonase (Sigma), and 1? cOmplete Mini EDTA-free Protease Inhibitor Cocktail (Roche). Lysates were clarified by centrifugation (15,000?g, 20 min, 4? C.) and the total protein content of the supernatant was quantified using a BCA assay. Samples were prepared using equal amounts of total protein and LDS sample buffer (Invitrogen). Samples were resolved by SDS-PAGE using NuPAGE 4-12% Bis-Tris midi gels (Invitrogen, followed by transfer to Amersham Protran 0.45 NC nitrocellulose membrane (GE Healthcare) using wet transfer. Precision Plus Protein All Blue (Bio-Rad) protein ladder was used as a standard. Membranes were blocked with 5% powdered skimmed milk (Marvel) in Tris-buffered saline with 0.1% Tween-20 (TBST), or 5% PhosphoBlocker (Cell BioLabs). Blots were probed (overnight at 4? C.) using the following primary antibodies (diluted in 5% BSA in TBST) as appropriate: panKras (Sigma, SAB1404011, 1:2,000), panKras (Abcam, ab275876, 1:1,000), p44/42 Erk1/2 (AF647 conjugate, Cell Signaling Technologies 5376, 1:2,000), phosphor-p44/42 Erk1/2 (Thr202/Tyr204) (AF488 conjugate, Cell Signaling Technologies 13214, 1:1,000). The next day, blots were washed with TBST and incubated (1 h at RT) with anti-Tubulin hFAB-rhodamine (BioRad, 12004166. 1:10,000) and and anti-GAPDH hFAB-rhodamine (BioRad, #12004168, 1:10,000) primary antibodies, plus either anti-rabbit IRDye 800CW (Licor 1:10,000), anti-Rabbit StarBright Blue 700 (BioRad, 1:10,000), Goat Anti-Mouse StarBright Blue 700 (BioRad, 1:10,000) or GoatAnti-Mouse StarBright Blue 520 (BioRad, 1:10,000) secondary antibody as appropriate. Blots were visualised using a Bio-Rad ChemiDoc MP Imaging System, and band quantification was performed using Image Studio software (LiCor). Band intensities were normalized to the loading controls and reported as % of the average 0.1% DMSO vehicle intensity. Degradation data was plotted and analysed using Prism (Graphpad, version 8).

[0926] 1.7 Assay 7Degradation of PARP1 by Capillary Electrophoresis.

[0927] HCC1937 cells were seeded (0.5 million cells/well) in 24-well plates overnight prior to 24 h treatment with test compounds at the desired concentration (DMSO final concentration 0.2%). After 24 h incubation, cells were washed with PBS and lysed with RIPA buffer (Sigma) containing 1? cOmplete Mini EDTA-free Protease Inhibitor Cocktail (Roche). Lysates were clarified by centrifugation (10,000 rpm, 10 min, 4? C.) and the total protein content of the supernatant was quantified using a BCA assay. Capillary-based immunoassays were performed using a standard WES (Simple Western) protocol (ProteinSimple). Lysates were loaded onto WES plates at 1.5 ?g/well total protein. The following antibodies and antibody concentrations were used: Anti-PARP(CST #9532, 1:250 dilution), Anti-Tubulin (CST #2125, 1:250 dilution), secondary Anti-Rabbit(CST #7074S, 1:500 dilution). Data was produced by the WES Compass software as chemiluminescent counts and displayed as an electropherogram, and the chemiluminescent peak area value was used for all calculations. The amount of target protein was normalized to the loading control and reported as % of the average 0.1% DMSO vehicle intensity. Degradation data was plotted and analysed using Prism (Graphpad, version 8).

[0928] 1.8 Assay 8Degradation of Mutant EGFR by Immunoblot.

[0929] NCI-H1975 (1.5?10.sup.5) cells were seeded in 12-well plates (1 mL medium) overnight prior to 24 h treatment with test compounds at the desired concentration (DMSO final concentration 0.1%). After 24 h incubation, cells were washed with PBS and lysed with RIPA buffer (Sigma) supplemented with 1? protease inhibitor cocktail (Roche) and 1 U/mL Benzonase (Merck). Lysates were clarified by centrifugation (17,000?g, 20 min, 4? C.) and the total protein content of the supernatant was quantified using a BCA assay. Samples were prepared using equal amounts of total protein and LDS sample buffer (Invitrogen). Samples were resolved by SDS-PAGE using NuPAGE 4-12% Bis-Tris midi gels (Invitrogen, followed by transfer to Amersham Protran 0.45 NC nitrocellulose membrane (GE Healthcare) using wet transfer. Precision Plus Protein All Blue (Bio-Rad) protein ladder was used as a standard. The membrane was blocked with 5% powdered skimmed milk (Marvel) in Tris-buffered saline with 0.1% Tween-20 (TBST). Blots were probed (overnight at 4? C.) using the following primary antibodies (diluted in 5% BSA in TBST) as appropriate: total EGFR (Cell Signaling Technology, CST #4267, 1:2000), L858R EGFR (Cell Signaling Technology, CST #3197, 1:1000) and phospho-EGFR (Cell Signaling Technology, CST #3777,1:2000). The next day, blots were washed with TBST and incubated (1 h at RT) with anti-Tubulin hFAB-rhodamine (BioRad, 12004166. 1:20000) and and anti-GAPDH hFAB-rhodamine (#12004168, 1:20000) primary antibodies, plus either anti-rabbit IRDye 800CW (Licor 1:20,000 dilution) or anti-mouse IRDye 800CW (Licor 1:20,000 dilution) secondary antibody. Blots were visualised using a Bio-Rad ChemiDoc MP Imaging System, and band quantification was performed using Image Studio software (LiCor). Band intensities were normalized to loading controls and reported as % of the average 0.1% DMSO vehicle intensity. Degradation data was plotted and analysed using Prism (Graphpad, version 8).

Example 1

[0930] The impact on cell viability of MV4;11 cells was evaluated according to the procedure outlined in assay 3 for compounds A1, A2, A8 and A9. The IC.sub.50-values are shown in table 1 below.

TABLE-US-00020 Compound No. IC.sub.50 (nM) A1 48 A2 2 A8 1 A9 3

[0931] Table 1 shows IC.sub.50 parameters for compounds A1, A2, A8 and A9.

[0932] It was surprisingly observed that modifications to the amido group on the warhead (as present in A2, A8 and A9) gave significant improvements in terms of the anti-proliferative activity (cell potency) of the bifunctional molecule as evidenced by their significantly lower IC.sub.50 values.

[0933] The enhanced cell potency of these compounds has been shown to correlate well with significantly improved BET degradation, particularly lower DC.sub.50 and higher D.sub.max parameters. For example, the correlation between the IC.sub.50 and DC.sub.50 values for a number of bifunctional molecules is shown in FIG. 1.

Example 2Efficacy of A2 in a Cancer Cell Panel

[0934] Cancer cell panel screening was provided as a service from OncoLead GmbH & Co. KG. After a lag phase of 48 h, each cell line was treated with six different concentrations (10.sup.?10, 10.sup.?9, 10.sup.?8, 10.sup.?7, 10.sup.?6, 10.sup.?5 M) of either A2 or I-BET726 for 72 h. Concentrations to give half-maximal growth inhibition (GI.sub.50) were determined using the Sulforhodamine B method. Log(GI.sub.50) values were plotted as single points (I-BET726, cross; A2, dot) superimposed on the bar graph and plotted along the right y axis. Log ratio of the GI.sub.50 determined for I-BET726 versus the GI.sub.50 determined for A2 were plotted for each cell line tested (bars, left y axis).

[0935] Values>0 indicate cell lines where BET-degradation by A2 shows greater efficacy than the inhibitor I-BET726 due to catalytic activity, whereas values<0 indicate cell lines where BET degrader A2 is less efficacious than BET-inhibition with I-BET726. Where no specific GI.sub.50 value could be determined, a GI.sub.50 of at least 10.sup.?5 M was assumed to calculate the inhibitor vs degrader ratio.

[0936] The results are illustrated on FIG. 2 and show that A2 shows a broad efficacy against a wide range of tumour cell lines.

[0937] Similar studies have previously been carried out on CRBN PROTAC (dBET6) and VHL PROTAC (MZ1) (see, for example, Ottis et al, ACS Chem. Biol. 2019, 14, 2215-2223 under identical assay conditions). A comparison of these results is shown in table 2 below.

TABLE-US-00021 GI.sub.50 comparison (BET degrader: up to 94 tumor cell lines) CRBN (dBET6) vs VHL (MZ1) vs A2 # tumor cell lines Highly Low sensitivity/ Degrader Treatment sensitive Resistant GI.sub.50, <1 ?M CRBN PROTAC (dBET6) 29% 52% 58% VHL PROTAC (MZ1) 47% 26% 83% A2 66% 15% 100%

[0938] Table 2 shows GI.sub.50 comparison (BET degrader: up to 94 tumor cell lines) for CRBN (dBET6), VHL (MZ1) and A2. Highly sensitive defined as degrader GI.sub.50>3 fold more potent than inhibitor. Low sensitivity/resistant defined as degrader GI.sub.50<inhibitor. Other cell lines showed intermediate activity.

[0939] The data demonstrate that A2 shows a broader range of efficacy across the tumour cell lines tested in comparison to the CRBN PROTAC degrader (dBET6) and VHL PROTAC degrader (MZ1).

[0940] Further N-alkylated warheads were then investigated to determine their ability to promote selective protein degradation in two test systems (one in which the target protein was BRD4 and the other in which the target protein was the kinase CDK9).

Example 3aBRD4 Degradation

[0941] The degradation of target protein BRD4 was detected according to the procedure outlined in assay 1 for the following compounds.

TABLE-US-00022 No. Structure A2 [00409] embedded image A3 [00410] A4 [00411] A5 [00412] A6 [00413] A7 [00414] A8 [00415] A9 [00416]

[0942] Table 3 shows the bifunctional molecules that were analysed in accordance with the procedure outlined in assay 1.

[0943] The DC.sub.50 values for compounds A2 to A5 and A7 to A9 were found to be less than 1000 nM. The DC.sub.50 for compound A6 was found to be less than 10000 nM. These molecules are all considered to be effective degraders.

Example 4

[0944] The degradation of target protein CDK9 was detected according to the procedure outlined in assay 2 for the following compounds.

TABLE-US-00023 Compound no. Structure A10 [00417] embedded image A11 [00418] A12 [00419] A13 [00420] A14 [00421] A15 [00422] A16 [00423] A17 [00424] A18 [00425] A19 [00426] A20 [00427] A21 [00428] A22 [00429] A23 [00430] A24 [00431] A25 [00432] A26 [00433] A27 [00434] A28 [00435] A29 [00436] A30 [00437] A31 [00438] A32 [00439] A33 [00440] A34 [00441] embedded image A35 [00442] A36 [00443] A37 [00444] A38 [00445] A39 [00446]

[0945] Table 4 shows the bifunctional molecules that were analysed in accordance with the procedure outlined in assay 2, in particular to determine the residual CDK9 abundance after 8 h of treatment with 100 nM of the bifunctional molecule.

[0946] In all cases, the residual CDK9 abundance after 8 h treatment with 100 nM of the bifunctional molecule was found to be less than 70%. Thus, all the above bifunctional molecules are considered to be effective degraders.

Example 5

[0947] The degradation of target protein CDK9 was detected according to the procedure outlined in assay 4 for the following compounds.

TABLE-US-00024 A44 [00447] embedded image A57 [00448] A58 [00449] A45 [00450] A46 [00451] A47 [00452] A48 [00453] A49 [00454] A50 [00455] A51 [00456] A56 [00457] A54 [00458] A59 [00459] A53 [00460] A55 [00461] A60 [00462]

[0948] The DC.sub.50 values for compounds in table 5 were found to be less than 1000 nM. These molecules are all considered to be effective degraders.

Example 6

[0949] The ability of compounds to demonstrate in vivo systemic drug exposure after oral dosing in rodents was assessed using the following protocol:

[0950] Test compound was administered to C57BL/6 mice, (6-8 weeks, 18-20 g, female, N=6, purchased from JH Laboratory Animal Co. with free access to food and water) at the indicated dose level via oral gavage (p.o., 10 mL/kg, 5% DMSO+95%(15% HP-?-CD in Water) vehicle, oral dosing leg) or tail vein (i.v., 5 mL/kg, 5% DMSO+95%(15% HP-?-CD in Water) vehicle, intravenous leg). The animal was restrained manually at designated time points up to 24 h, approximately 110 ?L of blood sample was collected via facial vein into K2EDTA tubes. Blood sample was put on ice and centrifuged at 2000 g for 5 min to obtain plasma sample within 15 minutes which was analysed on an LC-MS/MS-19 (Triple Quad 5500) in positive ion ESI mode following sample prep and HPLC elution:

[0951] Mobile Phase: [0952] Mobile Phase A: H.sub.2O0.025% FA1 mM NH.sub.4OAC [0953] Mobile Phase B: MeOH0.025% FA1 mM NH.sub.4OAc

TABLE-US-00025 Time (min) Mobile Phase B (%) 0.20 5 0.50 95 1.30 95 1.31 5 1.80 stop [0954] Column: ACQUITY UPLC BEH C18 2.1*50 mm 1.7 um [0955] Flow rate: 0.60 mL/min [0956] Column temperature: 60? C.

[0957] Example A37 showed AUC.sub.(0-INF) 841 hr*ng/mL following an intravenous 1 mg/kg dose and AUC.sub.(0-INF) 605 hr*ng/mL following an oral 10 mg/kg dose to give an oral bioavailability of 7.2%

Example 7

[0958] The ability of compounds to demonstrate in vivo drug exposure in the brain after systemic dosing in rodents was assessed using the following protocol:

[0959] Test compound was administered to C57BL/6 mice, (6-8 weeks, 18-20 g, female, N=6, purchased from JH Laboratory Animal Co. with free access to food and water) at the indicated dose level via tail vein (i.v., 5 mL/kg, 5% DMSO+95%(15% HP-?-CD in Water) vehicle).

[0960] Blood collection: The animal was restrained manually at designated time points up to 24 h, approximately 110 ?L of blood sample was collected via facial vein into K2EDTA tubes. Blood sample was put on ice and centrifuged at 2000 g for 5 min (4? C.) to obtain plasma sample within 15 minutes.

[0961] Brain collection: The animal was euthanized under CO.sub.2. A mid-line incision was made in the animal's scalp and skin retracted. The skull overlying the brain was removed. The whole brain was collected, rinsed with cold saline, dried on filtrate paper, weighted, snap frozen by placing into dry-ice. The sample was homogenized with 3 volumes (v/w) of PBS prior to analysis. Samples were analysed on an LC-MS/MS-19 (Triple Quad 5500) in positive ion ESI mode following sample prep and HPLC elution:

[0962] Mobile Phase: [0963] Mobile Phase A: H.sub.2O0.025% FA1 mM NH.sub.4OAC [0964] Mobile Phase B: MeOH0.025% FA1 mM NH.sub.4OAc

TABLE-US-00026 Time (min) Mobile Phase B (%) 0.20 5 0.60 95 1.20 95 1.21 5 1.80 stop [0965] Column: waters BEH C18 (2.1?50 mm, 1.7 ?m) [0966] Flow rate: 0.60 mL/min [0967] Column temperature: 60? C.

[0968] Example A39 showed a plasma AUC.sub.(0-INF) 1336 hr*ng/mL and brain AUC.sub.(0-INF) 845 hr*ng/mL following an intravenous 5 mg/kg dose showing a high level of brain exposure (brain:plasma 0.6).

[0969] This high level of brain exposure is in contrast to many other types of bifunctional degraders, such as CRBN and VHL PROTACs, which do not routinely allow for CNS (central nervous system) penetration.

Example 8

[0970] The degradation of target protein PARP1 was detected according to the procedure outlined in assay 7 for the following compounds: A61, A62, A63.

[0971] In all cases, the residual PARP1 abundance after 24 h treatment with 100 nM of the bifunctional molecule was found to be less than 50%. Thus, all the above bifunctional molecules are considered to be effective degraders.

Example 9

[0972] The degradation of target protein mutant KRas (G12C) was detected according to the procedure outlined in assay 8 for the compound A64.

[0973] The residual KRas (G12C) abundance after 24 h treatment with 1000 nM of the bifunctional molecule was found to be less than 50%. Thus, the bifunctional molecule is considered to be an effective degrader.

Example 10

[0974] The degradation of target protein BRD9 was detected according to the procedure outlined in assay 5 for the compounds A65, A66.

[0975] The residual BRD9 abundance after 24 h treatment with 100 nM of the bifunctional molecule was found to be less than 50%. Thus, the above bifunctional molecules are considered to be effective degraders.

Example 11

[0976] The degradation of target protein mutant EGFR (L858R) was detected according to the procedure outlined in assay 8 for the compound A67.

[0977] The residual EGFR (L858R) abundance after 24 h treatment with 1000 nM of the bifunctional molecule was found to be less than 50%. Thus, the bifunctional molecule is considered to be an effective degrader.

Novel Bifunctional Molecules For Targeted Protein Degradation

Inventors

Cpc classification

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C07D417/06

CHEMISTRY; METALLURGY

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A61K47/545

HUMAN NECESSITIES

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C07D277/10

CHEMISTRY; METALLURGY

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G01N2500/10

PHYSICS

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G01N33/5011

PHYSICS

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A61P35/00

HUMAN NECESSITIES

International classification

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A61K47/54

HUMAN NECESSITIES

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A61P35/00

HUMAN NECESSITIES

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G01N33/50

PHYSICS

Abstract

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