EP4076659A1 - Macrocyclic amides acting as plasmepsin inhbitors for the treatment of malaria - Google Patents

Abstract

The present invention relates to medicine and in particular to the treatment of malarial infections, more particularly to inhibitors of malarial aspartic proteases - plasmepsins. Even more particularly, the invention relates to macrocyclic amides and pharmaceutical compositions thereof and their use as inhibitors for plasmepsins (Plm).

Description

MACROCYCLIC AMIDES ACTING AS PLASMEPSIN INHBITORS FOR THE TREATMENT OF MALARIA Field of invention [001] The present invention relates to medicine, and in particular to the treatment of malarial infections, more particularly to inhibitors of malarial aspartic proteases - plasmepsins. Even more particularly, the invention relates to macrocyclic amides and pharmaceutical compositions thereof and their use as inhibitors for plasmepsins (Plm). Background of invention [002] Widespread resistance to practically all currently used drugs has stimulated the search for antimalarials with novel mechanisms of action (Hyde, J. E. Drug-resistant malaria - an insight. FEBS J. 2007, 274, 4688.; Choi, S. R.; Mukherjee, P.; Avery, M. A. The fight against drug-resistant malaria: novel plasmodial targets and antimalarial drugs. Curr. Med. Chem. 2008, 15, 161; Wells, T. N.; Alonso, P. L.; Gutteridge, W. E. New medicines to improve control and contribute to the eradication of malaria. Nat. Rev. Drug Discov. 2009, 8, 879.). [003] Resistance to current anti-malarial agents in malaria-endemic regions continues to spread, indicating that current therapeutic agents will be practically ineffective in the near future. A precondition for the development of new antimalarial agents is inhibition of the malaria parasite life cycle through a mechanism that differs from the mode of action of currently used therapeutic agents (N. K. Sahu, S. Sahu and D. V. Kohli, Novel Molecular Targets for Antimalarial Drug. Chem. Biol. Drug. Des., 2008, 71, 287.) [004] This could be achieved by targeting functional malarial proteins involved in the blood stage of the parasite life cycle. Malarial enzymes such as malarial aspartic proteases - plasmepsins (Plms) have been recognized as promising molecular targets for new drug development (Meyer, M. J.; Goldberg, D. E. Recent advances in plasmepsin medicinal chemistry and implications for future antimalarial drug discovery efforts. Curr. Top. Med. Chem. 2012, 12, 445.; Kumar Singh, A.; Rajendran, V.; Singh, S.; Kumar, P.; Kumar, Y.; Singh, A.; Miller, W.; Potemkin, V.; Poonam; Grishina, M.; Gupta, N.; Kempaiah. P.; Durvasulak, R.; Singh, B. K.; Dunn, B. M.; Rathi, B. Antiplasmodial activity of hydroxyethylamine analogs: Synthesis, biological activity and structure activity relationship of plasmepsin inhibitors. Bioorg. Med. Chem. 2018, 26, 3837.; Bobrovs, R.; Jaudzems, K.; Jirgensons, A. Exploiting Structural Dynamics To Design Open-Flap Inhibitors of Malarial Aspartic Proteases. J. Med. Chem., 2019, 62, 8931; Hodder, A. N.; Sleebs, B. E.; Czabotar, P. E.; Gazdik, M.; Xu, Y.; O’Neill, M. T.; Lopaticki, S.; Nebl, T.; Triglia, T.; Smith, B. J.; Lowes, K.; Boddey, J. A.; Cowman, A. F. Structural basis for plasmepsin V inhibition that blocks export of malaria proteins to human erythrocytes. Nat. Struct. Mol. Biol. 2015, 22, 590). [005] Despite number of plasmepsin inhibitors reported, none of them have advanced to clinical trial. Summary of the invention [006] In a first aspect, the invention features a method of treating malarial infections in humans or animals, comprising administering to a human or animal in need of a therapeutically effective amount of a compound or prodrug thereof, or pharmaceutically acceptable salt, hydrate, solvate, or polymorph of said compound or prodrug, wherein the compound is an inhibitor of malarial aspartic protease - plasmepsin (Plm) [007] In another aspect, the invention features a pharmaceutical composition for treatment of malaria infections comprising a therapeutically effective amount of a composition comprising (i) a compound or prodrug thereof, or pharmaceutically acceptable salt, hydrate, solvate, or polymorph of said compound or prodrug; and (ii) a pharmaceutically acceptable carrier, wherein the compound is an inhibitor of malaria aspartic proteases - plasmepsins (Plms). [008] In another aspect, the invention features the use of a compound or prodrug thereof, or pharmaceutically acceptable salt, hydrate, solvate, or polymorph of said compound or prodrug, wherein the compound is an inhibitor of malaria aspartic proteases - plasmepsins (Plms), in the manufacture of a medicament for treatment or prevention of malaria infections. [009] In another aspect, the invention features a compound or prodrug thereof, or pharmaceutically acceptable salt or ester of said compound or prodrug for use in treating or preventing malaria infections, wherein the compound is an inhibitor of malaria aspartic proteases - plasmepsins (Plms). [010] In one embodiment an inhibitor of malaria aspartic protease - plasmepsin (Plms) is a compound of Formula I, generally referred herein as macrocyclic amides: [011] General formula I [012] wherein: R¹, R², R³ are independently H, C_1-6alkyl, cycloC_3-12alkyl, cycloC_3-12alkyl-C_1-6alkyl, C_{2- 6}alkenyl, C_2-6alkynyl, aryl, biaryl, arylC_1-6alkyl, arylC_2-6alkenyl, arylC_2-6alkynyl, heteroaryl, heteroarylC_1-6alkyl, heteroarylC_2-6alkenyl, R⁴O(CH₂)_n, R⁴S(CH₂)_n, R⁴OC(=O)(CH₂)_n, R⁴N(R⁵)C(=O)(CH₂)_n, R⁴N(R⁵)(CH₂)_n, -F, -Cl, -Br, -I, -CF₃, -CH₂CF₃, -CF₂CF₂H, -OH, -L-OH,-O-L-OH, -OR⁴, -O-L-NH₂, -O-L-NHR⁴, -O-L-N(R⁴)R⁵, -L-OR⁴,-O-L-OR⁴,-OCF₃, -OCH₂CF₃, -OCF₂CF₂H, -L-OR⁴,-O-L-OR⁴,-OCF₃, -OCH₂CF₃, -OCF₂CF₂H, SR⁴, SCF₃, - CN, -NO₂, -NO₂, -NH₂, -NHR⁴, -NR⁴ 2, -N(R⁴)R⁵, -L-NH₂, -L-NHR⁴, -L-NR⁴ 2, -L-N(R⁴) R⁵,-NH-L-NH₂, -NH-L-NHR⁴, -NH-L-N(R⁴)R⁵, -NH-L-N(R⁴)R⁵,-NR⁴-L-NH₂, -NR⁴-L-NHR⁴, -NR⁴-L-N(R⁴)R⁵, -NR⁴-L-N(R⁴)R⁵, -N(R⁴)R⁵, -C(=O)OH, -C(=O)OR⁴, -C(=O)NH₂, -C(=O)NHR⁴, -C(=O)N(R ⁴)R⁵, -C(=O)N(R⁴)R⁵,-NHC(=O)R⁴, -NR⁴C(=O)R⁵, -NHC(=O)OR⁴, -NR⁴C(=O)OR⁵, -OC(=O)NH₂, -OC(=O)NHR⁴, -OC(=O)N(R⁴)R⁵, -OC(=O) R⁴NR⁵,-OC(=O)R⁴, -C(=O)R⁴,-NHC(=O)NH₂, -NHC(=O)NHR⁴, -NHC(=O)NR⁴ 2, -NHC(=O)N(R⁴)R⁵, -NR⁴C(=O)NH₂, -N(R⁴)C(=O)NHR⁵, -NR⁴C(=O)N (R⁴)R⁵ [013] wherein: n is an integer selected from 1 to 6; L represents -W^A-X^A-Y^A-Z^A-; or -W^A-X^A-Y^A-, or -W^A-X^A- or -W^A- wherein: W^A- represents a single bond, oxygen, sulfur, -NR⁶ or –CR⁶R⁷, X^A represents oxygen, sulfur, -NR⁶or –C(R⁶)R⁷, Y^A represents oxygen, sulfur, -NR⁶ or –C(R⁶)R⁷, Z^A represents oxygen, sulfur, -NR⁶ or –C(R⁶)R⁷; [014] wherein: R⁶ and R⁷ are independently H, C_1-6alkyl, cycloC_3-12alkyl, cycloC_3-12alkyl- C_1-6alkyl, C_{2- 6}alkenyl, C_2-6alkynyl, aryl, biaryl, aryl C_1-6alkyl, arylC_2-6alkenyl, arylC_2-6alkynyl, heteroaryl, heteroarylC_1-6alkyl, heteroarylC_2-6alkenyl, heteroarylthio, 2,3-dihydro- 1H-indenyl, C_{1- 6}alkoxyC_1-6alkyl, aryloxyarylC_1-6alkoxy, C_1-6alkylthio, C_4-6alkenylthio, cycloC_3-12alkylthio, cycloC_3-12alkyl-C_1-6alkylthio, cycloC_3-12alkyl-C_3-6alkenylthio, C_1-6alkoxyC_1-6alkylthio, C_{1- 6}alkoxyC_3-6alkenylthio, arylC_3-6alkenylthio, heteroarylC_1-6alkylthio, C_1-6alkylsulfonyl, cycloC_3-12alkyl-C_1-6alkylsulfonyl, arylC_1-6alkylsulfonyl, C_1-6alkylamino, di-C_1-6alkylamino, cycloC_3-12alkylamino, C_1-C₆alkoxy-cycloC_3-12 alkylamino, cycloC_3-12alkyl-C_1-6alkylamino, di-C_1-6alkylaminoC_1-6alkyl, C_1-6alkoxy-C_2-6alkylamino, arylamino, arylC_1-6alkylamino, N- cycloC_3-12alkyl-N-C_1-6alkylamino, N-aryl-N- C_1-6alkylamino, N-arylC_1-6alkyl-N- C_{1- 6}alkylamino, 2-indanylamino, tetrahydrofuryl, pyrrolidino, piperidino, 4-arylpiperidino, 4- heteroarylpiperidino, morpholino, piperazino, 4-C_1-6alkylpiperazino, 4-arylpiperazino, hexamethyleneimino, benzazepinyl, l,3-dihydro-2H-isoindol-2-yl, heteroarylC_1-6alkoxy, heteroarylamino, or heteroarylC_1-6alkylamino.

[015] Q represents -A-D-E-G-H-J-; or -A=D-E-G-H-J-; or -A-D=E-G-H-J-; or -A-D-E=G- H-J-; or -A-D-E-G= H-J-; or -A-D-E-G-H=J-; or -D-E-G-H-J-; or -D=E-G-H-J-; or -D-E=G-H-J-; or -D-E-G=H-J-; or -D-E-G-H=J-; or -E-G-H-J-; or -E=G-H-J-; or -E-G=H-J-; or -E-G-H=J-; or -G-H-J-; or -G=H-J-; or -G-H=J-; or -H-J-; or -H=J-; or -J-

[016] wherein:

A represents, oxygen, sulfur, -NR⁸ or -CR⁸R⁹ or =CR⁸,

D represents oxygen, sulfur, -NR¹⁰ or -CR¹⁰R¹¹ or =CR¹⁰,

E represents oxygen, sulfur, -NR¹² or -CR¹²R¹³ or =CR¹²,

G represents oxygen, sulfur, -NR¹⁴ or -CR¹⁴R¹⁵ or =CR¹⁴,

H represents oxygen, sulfur, -NR¹⁶or -CR¹⁶R¹⁷ or =CR¹⁶,

J represents oxygen, sulfur, -NR¹⁸ or-CR¹⁸R¹⁹ or=CR¹⁸,

R⁸, R⁹, R¹⁰, R¹¹,R¹², R¹³, R¹⁴, R¹⁵, R¹⁶ R¹⁷, R¹⁸, R¹⁹ are independently H, C_1-6alkyl, cycloC_{3- 12}alkyl, cycloC_3-12alkyl-C_1-6alkyl, C_2-6alkenyl, C_2-6alkynyl, aryl, biaryl, arylC_1-6alkyl, arylC_{2- 6}alkenyl, arylC_2-6alkynyl, heteroaryl, heteroarylC_1-6alkyl, heteroarylC_2-6alkenyl, heteroarylthio, 2,3 -dihydro- lH-indenyl, C_1-6alkoxy C_1-6alkyl, aryloxyaryl C_1-6alkoxy, C_{1- 6}alkylthio, C_4-6alkenylthio, cycloC_3-12alkylthio, cycloC_3-12alkyl-C_1-6alkylthio, cycloC_3-12alkyl- C_3-6alkenylthio, C_1-6alkoxyC_1-6alkylthio, C_1-6alkoxyC_3-6alkenylthio, arylC_3-6alkenylthio, heteroarylC_1-6alkylthio, C_1-6alkylsulfonyl, cycloC_3-12alkyl-C_1-6alkylsulfonyl, arylC_{1- 6}alkylsulfonyl, C_1-6alkylamino, di-C_1-6alkylamino, cycloC_3-12alkylamino, C₁-C₆alkoxy- cycloC₃-C₁₂alkylamino, cycloC_3-12alkyl-C_1-6alkylamino, di-C_1-6alkylaminoC_1-6alkyl, C_{1- 6}alkoxy-C_2-6alkylamino, arylamino, arylC_1-6alkylamino, N-cycloC_3-12alkyl-N-C_1-6alkylamino, N-aryl-N-C_1-6alkylamino, N-arylC_1-6alkyl-N-C_1-6alkylamino, 2-indanylamino, tetrahydrofuryl, pyrrolidino, piperidino, 4-arylpiperidino, 4-heteroarylpiperidino, morpholino, piperazino, 4-C_1-6alkylpiperazino, 4-arylpiperazino, hexamethyleneimino, benzazepinyl, 1,3-dihydro-2H-isoindol-2-yl, heteroarylC_1-6alkoxy, heteroarylamino, or heteroarylC_{1- 6}alkylamino; or R⁸ and R⁹; R¹⁰ and R¹¹ _; R¹² and R¹³; R¹⁴, and R¹⁵ _; R¹⁶ and R¹⁷, R¹⁸ and R¹⁹ are independently =O,=NR²⁰ or =NOR²⁰; [017] wherein: R²⁰ are independently H, C_1-6alkyl, cycloC_3-12alkyl, cycloC_3-12alkyl-C_1-6alkyl, C_2-6alkenyl, C_{2- 6}alkynyl, aryl, biaryl, arylC_1-6alkyl, arylC_2-6alkenyl, arylC_2-6alkynyl, heteroaryl, heteroarylC_1-6alkyl, heteroarylC_2-6alkenyl [018] wherein:

R⁸, R⁹, R¹⁰, R¹¹,R¹², R¹³, R¹⁴, R¹⁵, R¹⁶ R¹⁷, R¹⁸, R¹⁹ R⁸ and R¹⁰; R⁸ and R¹²; R⁸ and R¹⁴ _; R⁸ and R¹⁶; R⁸ and R¹⁸; R¹⁰ and R¹²; R¹⁰ and R¹⁴ _; R¹⁰ and R¹⁶; R¹⁰ and R¹⁸; R¹² and R¹⁴ _; R¹² and R¹⁶; R¹² and R¹⁸; R¹⁴ and R¹⁶; R¹⁴ and R¹⁸ R¹⁶ and R¹⁸ taken together represent -W^B-X^B-Y^B-Z^B-, or -W^B-X^B-Y^B-, or -W^B-X^B- or -W^B- [019] wherein: W^B represents a single bond, oxygen, sulfur, -NR²¹ or –CR²¹R²², X^B represents oxygen, sulfur, -NR²¹ or –C(R²¹)R²², Y^B represents oxygen, sulfur, -NR²¹ or –C(R²¹)R²², Z^B represents oxygen, sulfur, -NR²¹ or –C(R²¹)R²², [020] wherein: R²¹ and R²² are independently H, C_1-6alkyl, cycloC_3-12alkyl, cycloC_3-12alkyl-C_1-6alkyl, C_{2- 6}alkenyl, C_2-6alkynyl, aryl, biaryl, arylC_1-6alkyl, arylC_2-6alkenyl, arylC_2-6alkynyl, heteroaryl, heteroarylC_1-6alkyl, heteroarylC_2-6alkenyl, heteroarylthio, 2,3-dihydro-1H-indenyl, C_{1- 6}alkoxyC_1-6alkyl, aryloxyarylC_1-6alkoxy, C_1-6alkylthio, C_4-6alkenylthio, cycloC_3-12alkylthio, cycloC_3-12alkyl-C_1-6alkylthio, cycloC_3-12alkyl-C_3-6alkenylthio, C_1-6alkoxyC_1-6alkylthio, C_{1- 6}alkoxyC_3-6alkenylthio, arylC_3-6alkenylthio, heteroarylC_1-6alkylthio, C_1-6alkylsulfonyl, cycloC_3-12alkyl-C_1-6alkylsulfonyl, arylC_1-6alkylsulfonyl, C_1-6alkylamino, di-C_1-6alkylamino, cycloC_3-12alkylamino, C₁-C₆alkoxy-cycloC₃-C₁₂alkylamino, cycloC_3-12alkyl-C_1-6alkylamino, di-C_1-6alkylaminoC_1-6alkyl, C_1-6alkoxy-C_2-6alkylamino, arylamino, arylC_1-6alkylamino, N- cycloC_3-12alkyl-N-C_1-6alkylamino, N-aryl-N-C_1-6alkylamino, N-arylC_1-6alkyl-N-C_{1- 6}alkylamino, 2-indanylamino, tetrahydrofuryl, pyrrolidino, piperidino, 4-arylpiperidino, 4- heteroarylpiperidino, morpholino, piperazino, 4-C_1-6alkylpiperazino, 4-arylpiperazino, hexamethyleneimino, benzazepinyl, 1,3-dihydro-2H-isoindol-2-yl, heteroarylC_1-6alkoxy, heteroarylamino, or heteroarylC_1-6alkylamino; [021] and optical isomers, pharmaceutically acceptable salts, hydrates, solvates, and polymorphs thereof. [022] In one embodiment, the treatment is treatment of a disease or disorder that is mediated by a malarial aspartic protease or human aspartic protease. [023] In one embodiment, the treatment is treatment of a disease or disorder that is ameliorated by the inhibition of a malarial aspartic protease or human aspartic protease. [024] In one embodiment, the treatment is treatment of a disease or disorder that is treated by a malarial aspartic protease or human aspartic protease inhibitor. [025] In another aspect, the invention features a kit comprising a macrocyclic amide described herein, preferably provided as a pharmaceutical composition and in a suitable container and/or with suitable packaging. [026] In another aspect, the invention features compounds obtainable by a method of synthesis as described herein, or a method comprising a method of synthesis as described herein. [027] In another aspect, the invention features compounds obtained by a method of synthesis as described herein, or a method comprising a method of synthesis as described herein. [028] In another aspect, the invention features novel intermediates, as described herein, which are suitable for use in the methods of synthesis described herein. [029] In another aspect, the invention features the use of such novel intermediates, as described herein, in the methods of synthesis described herein. [030] As will be appreciated by one of skilled in the art, features and preferred embodiments of one aspect of the invention will also pertain to other aspects of the invention. Description of invention [031] Malarial aspartic proteases - plasmepsins (Plms) which have been identified as a group of promising biological targets for the development of new anti-malarial agents (Meyer, M. J.; Goldberg, D. E. Recent advances in plasmepsin medicinal chemistry and implications for future antimalarial drug discovery efforts. Curr. Top. Med. Chem. 2012, 12, 445.; Kumar Singh, A.; Rajendran, V.; Singh, S.; Kumar, P.; Kumar, Y.; Singh, A.; Miller, W.; Potemkin, V.; Poonam; Grishina, M.; Gupta, N.; Kempaiah. P.; Durvasulak, R.; Singh, B. K.; Dunn, B. M.; Rathi, B. Antiplasmodial activity of hydroxyethylamine analogs: Synthesis, biological activity and structure activity relationship of plasmepsin inhibitors. Bioorg. Med. Chem. 2018, 26, 3837.; Bobrovs, R.; Jaudzems, K.; Jirgensons, A. Exploiting Structural Dynamics To Design Open-Flap Inhibitors of Malarial Aspartic Proteases. J. Med. Chem., 2019, 62, 8931.; Hodder, A. N.; Sleebs, B. E.; Czabotar, P. E.; Gazdik, M.; Xu, Y.; O’Neill, M. T.; Lopaticki, S.; Nebl, T.; Triglia, T.; Smith, B. J.; Lowes, K.; Boddey, J. A.; Cowman, A. F. Structural basis for plasmepsin V inhibition that blocks export of malaria proteins to human erythrocytes. Nat. Struct. Mol. Biol. 2015, 22, 590). [032] When testing the novel macrocyclic amides for their ability to inhibit Plm IV we have unexpectedly discovered, that said derivatives exhibit pronounced inhibitory properties toward said aspartic protease, and thus are useful in treatment of malaria. [033] According to this invention, the results from Plm inhibition studies demonstrate that macrocyclic amides are novel class inhibitors of plasmepsins. Several example compounds from the present invention display nanomolar to low micromolecular plasmepsin inhibitory potency Stereochemistry [034] Many of the chemical structures shown herein indicate one or more specific stereoisomeric configurations. Similarly, many of the chemical structures shown herein are silent in this respect, and do not indicate any stereoisomeric configuration. Similarly, many of the chemical structures shown herein indicate the specific stereoisomeric configurations at one or more positions, but are silent with respect to one or more other positions. Where a chemical structure herein is silent with respect to the stereoisomeric configuration at a position, that structure is intended to depict all possible stereoisomeric configurations at that position, both individually, as if each possible stereoisomeric configuration was individually recited, and also as a mixture (e.g., a racemic mixture) of stereoisomers. Combinations [035] Each and every compatible combination of the embodiments described above is explicitly disclosed herein, as if each and every combination was individually and explicitly recited. Examples of Specific Embodiments [036] The following examples further illustrate the invention, but should not be construed to limit the scope of the invention in any way. [037] The following macrocyclic amides 15 were prepared as examples of the current invention: General Synthesis [038] Principal building blocks 11a,b for the synthesis macrocyclic amides were prepared according to Scheme 1. N-Allyl-1,1-diphenylmethanimine (1) (prepared according to the literature procedure: J. Am. Chem. Soc. 2014, 136, 11256; Org. Lett., 2017, 19, 3490) was deprotonated and C-alkylated with commercially available benzylic chloride derivative. The resulting allyl amine derivative 2 was N-Boc protected and separated into enantiomers by using chiral chromatography. S-Enantiomer of allylamine 3 was subjected to asymmetric Sharpless dihydroxylation providing mixture of diastereomeric diols. The diastereomer 4 was separated by chromatography and transformed to epoxide 5 in Mitsunobu reaction conditions. Ring opening of epoxide 5 with amine gave intermediate 6 which was subjected to N- deprotection. The resulting amine 7 was N-coupled with benzene carboxylic acids 8a,b (8a is commercially available; 8b was prepared according to the Scheme 3) to give amides 9a,b. O- TBS protection provided intermediates 10a,b which were O-debenzylated in hydrogenolysis conditions to give building blocks 11a,b. These building blocks were used to prepare the target compounds 15 (Scheme 2). [039] O-alkylation of compounds 11a,b with alcohols 12a-d in Mitsunobu conditions provided intermediates 13. These were subjected to the cleavage of N-Nosyl group leading to amines 14. Hydrolysis of the ester group in intermediates 14 followed by macrolactamization and removal of O-TBS group resulted in target compounds 15. [040] Building block 8b was obtained according to Scheme 3. Commercially available iodoisophtalic acid monoester 16 was coupled with dipropylamine to give amide 17. This was subjected to metal-halogen exchange followed by carboxylation to give acid 8b. Protected amino alcohols 12a-d were prepared according to Scheme 4. Commercially available amino alcohols 18a-d were N-nosylated to give intermediates 19a-d which were N- alkylated with propylbromide. [041] Protected amino alcohol 12e was prepared from commercially available amine 20 (Scheme 5) which involved N-nosylation to give intermediate 21 which was N-alkylated with commercially available O-protected oxyalkylbromide, followed by O-desilylatation. Scheme 1 Scheme 3 Scheme 4 Scheme 5 [042] Synthesis of (±)-1-(3-(benzyloxy)phenyl)but-3-en-2-amine (2) Oven dried 250 mL round bottom flask was charged with the N-allyl-1,1- diphenylmethanimine (1) (11.3 g, 46.0 mmol), and it was dissolved in a dry THF (90 mL). The solution was cooled to -50°C in acetone/dry ice cooling bath. n-BuLi solution in THF (2.5 M, 18.38 mL, 46.0 mmol) was added dropwise in 15 min (colour changed to yellow, then greenish, then wine red). After stirring for 30 min, solution of the 3-benzyloxybenzylchloride (10.7 g, 46.0 mmol) in THF (45 mL) was added in 5 min (colour was still wine red). Reaction mixture was allowed to slowly heat up to -30°C and stirred for additional 1 h (colour changed to green-yellow). The reaction mixture was quenched with EtOH (yellow solution) then charged with 15 mL of 20% KHSO₄ solution (red suspension) and stirred overnight. MTBE was added to the mixture and the phases were separated. Aqueous phase was charged with NaOH solution, until it became cloudy, then it was extracted with DCM (x6). Combined organic phase (THF+MTBE+DCM) was washed with brine, dried over Na₂SO₄, filtered and evaporated. The residue was purified by column chromatography eluting with EtOAc+1% TEA). Product 2 (Rf = 0.15; PE : EtOAc, 1:1) was collected after evaporation as a colourless oil (7.14 g, 61%). [043] ¹H NMR (300 MHz, Chloroform-d) δ 7.50 – 7.28 (m, 5H), 7.25 – 7.13 (m, 1H), 6.92 – 6.75 (m, 3H), 5.88 (ddd, J = 17.0, 10.3, 6.3 Hz, 1H), 5.15 (dt, J = 17.2, 1.4 Hz, 1H), 5.08 – 5.01 (m, 3H), 3.60 (dddt, J = 7.9, 6.6, 5.4, 1.3 Hz, 1H), 2.81 (dd, J = 13.3, 5.4 Hz, 1H), 2.61 (dd, J = 13.3, 8.3 Hz, 1H). [044] Synthesis of tert-butyl (S)-(1-(3-(benzyloxy)phenyl)but-3-en-2-yl)carbamate (3) (±)-1-(3-(benzyloxy)phenyl)but-3-en-2-amine (2) (5.60 g, 22.1 mmol) and DIPEA (5.74 mL, 33.2 mmol) were dissolved in DCM (100mL), then Boc₂O (7.23 g, 33.2 mmol) solution in DCM (20 mL) was added within 1 minute. Mixture was stirred overnight at room temperature. (UPLC indicated full conversion of starting material). Solvent was evaporated and the residue was filtered through silica in small column, eluting with EtOAc. Solvent was evaporated and the residue was dissolved in 10% iPA/Heptane (20 mL). Heptane was added (50 mL) and the mixture was left at -20ºC overnight. The precipitate was filtered, washed with heptane and dried. The solid (6.8 g) was dissolved in 20 mL iPrOH with heating. Enantiomers of the product were separated using preparative HPLC (Conditions: ChiralPak ID column 250x30 mm - 5 um; 3%iPA/Heptane, 40 mL/min, ~RT=10.0 min (1V) and ~12.5 min (2V). Injection: 0.7-0.9 mL (about 200-250 mg of product average); each injection was performed after about 10 minutes (while 1V is eluting); 30 injections in total). Fractions with separated enantiomers were analysed to determine ee, % (Conditions: ChiralPak ID column; 2.5% iPA/Heptane, 1 mL/min, RT=13.0 min (1V) and 13.8 min (2V). Product 3 (2.6 g, 33%, ee 98%) was obtained as solid after evaporation of solvent. [045] ¹H NMR (400 MHz, Chloroform-d) δ 7.46 – 7.29 (m, 5H), 7.20 (dd, J = 8.2, 7.5 Hz, 1H), 6.87 – 6.77 (m, 3H), 5.78 (ddd, J = 17.1, 10.4, 5.1 Hz, 1H), 5.13 – 5.03 (m, 4H), 2.88 – 2.75 (m, 2H), 1.42 (s, 9H). [046] Synthesis of tert-butyl ((2S,3S)-1-(3-(benzyloxy)phenyl)-3,4-dihydroxybutan-2- yl)carbamate (4) A 500 mL round bottom flask was charged with tert-butyl (S)-(1-(3-(benzyloxy)phenyl)but-3- en-2-yl)carbamate (3) (2.60, 7.36 mmol), followed by t-BuOH (125 mL) and water (100 mL). AD-mix alfa (17.2 g) was added to the solution in 1 portion and the reaction mixture was stirred for 64 h at room temperature (UPLC full conversion, diastereomers (SS:SR, 6:4) Solid Na₂SO₃ (17.2 g) was added portionwise. After stirring for 30 min, EtOAc (100 mL) was added, phases were separated. Aqueous phase was washed with EtOAc (2 x 30 mL). Combined organic phase was washed with brine, dried over Na₂SO₄, filtered and evaporated. The residue was filtered through column filled with 40 mL of silica (washed with EtOAc). The collected eluate was evaporated. The residue was dissolved in DMSO (16 mL) and the diastereomers were separated by chromatography (conditions: Grace Prep GROM-SIL 120 ODS-5 ST, 10um (150x50mm column); 60 mL/min; injection volume 1.6mL; RT=25-26 min and 27-28 min). Product 4 (1.50 g, 53%, de=97%) was collected as colourless solid. [047] ¹H NMR (400 MHz, Chloroform-d) δ 7.47 – 7.29 (m, 5H), 7.25 – 7.20 (m, 1H), 6.85 (ddt, J = 12.5, 7.3, 1.3 Hz, 3H), 5.06 (s, 2H), 4.58 (d, J = 8.6 Hz, 1H), 3.83 (qd, J = 8.4, 4.4 Hz, 1H), 3.73 – 3.54 (m, 2H), 3.32 (d, J = 8.7 Hz, 1H), 3.05 (dd, J = 14.2, 4.3 Hz, 1H), 2.90 (dd, J = 14.3, 7.5 Hz, 1H), 1.39 (s, 9H). [048] HRMS [M+H]⁺: 410.1942, calcd.:410.1943 [049] Synthesis of tert-butyl ((S)-2-(3-(benzyloxy)phenyl)-1-((S)-oxiran-2- yl)ethyl)carbamate (5) An oven dried vial was charged with tert-butyl ((2S,3S)-1-(3-(benzyloxy)phenyl)-3,4- dihydroxybutan-2-yl)carbamate (4) (1.25 g, 3.23 mmol) and CHCl₃ (25 mL) was added. Triphenyl phosphine (0.846 mg, 3.23 mmol) was added to the solution in one portion followed by dropwise addition of diethyl azodicarboxylate (0.51 mL, 3.23 mmol). The reaction mixture was heated at 75^oC for 20 h (full conversion by UPLC). Reaction mixture was concentrated in vacuo and the crude product 5 was used in next step without further purification. [050] Synthesis of tert-butyl ((2S,3R)-1-(3-(benzyloxy)phenyl)-3-hydroxy-4-((2-(3- methoxyphenyl)propan-2-yl)amino)butan-2-yl)carbamate (6) Tert-butyl ((S)-2-(3-(benzyloxy)phenyl)-1-((S)-oxiran-2-yl)ethyl)carbamate (5) (1.19 g, 3.23 mmol) was dissolved in iPrOH (12 mL) and 2-(3-methoxyphenyl)propan-2-amine (2.13 g, 13.0 mmol) was added. The reaction mixture was heated at 70°C overnight and the solvent was evaporated. The residue was purified by column chromatography on silica gel eluting with diethyl ether. Product 6 (1.20 g, 70%) was collected as colourless oil. ¹H NMR (400 MHz, Chloroform-d) δ 7.45 – 7.28 (m, 5H), 7.25 – 7.15 (m, 2H), 7.03 – 6.96 (m, 2H), 6.86 – 6.73 (m, 4H), 5.04 (s, 2H), 4.57 (d, J = 9.2 Hz, 1H), 3.80 (s, 3H), 3.78 – 3.72 (m, 1H), 3.28 (dt, J = 7.6, 4.5 Hz, 1H), 2.92 (dd, J = 14.1, 4.7 Hz, 1H), 2.81 (dd, J = 14.2, 7.6 Hz, 1H), 2.48 – 2.34 (m, 2H), 1.45 (s, 6H), 1.37 (s, 9H). HRMS [M+H]⁺: 535.3179, calcd.: 535.3172 [051] Synthesis of (2R,3S)-3-amino-4-(3-(benzyloxy)phenyl)-1-((2-(3-methoxy phenyl)propan-2-yl)amino)butan-2-ol dihydrochloride (7) tert-Butyl ((2S,3R)-1-(3-(benzyloxy)phenyl)-3-hydroxy-4-((2-(3-methoxy phenyl)propan-2- yl)amino)butan-2-yl)carbamate (6) (1.69 g, 3.16 mmol) was dissolved in dioxane and then 4 M HCl (6.32 mL, 25.3 mmol) in 1,4-dioxane was added. The colourless solution was stirred at room temperature for 18 h then additional 4 M HCl (6.32 mL, 25.3 mmol) in 1,4-dioxane was added. The reaction mixture was stirred for another 6 h to achieve full conversion of starting material (UPLC monitoring). The yellowish solution was evaporated and the crude product 7 was used further without purification. [052] Synthesis of intermediates 9, general method A Exemplified for the synthesis of methyl 3-(((2S,3R)-1-(3-(benzyloxy)phenyl)-3-hydroxy-4- ((2-(3-methoxyphenyl)propan-2-yl)amino)butan-2-yl)carbamoyl) benzoate (9a) A round bottom flask was charged with mono-methyl isophthalate (8a) (569 mg, 3.16 mmol). Anhydrous DCM (50 mL) was added, and the resulting stirred suspension was cooled to 0°C. HOBt (666 mg, 3.48 mmol) was added followed by EDC (560 uL, 3.16 mmol) 20 min later. After 1 h, (2R,3S)-3-amino-4-(3-(benzyloxy)phenyl)-1-((2-(3-methoxyphenyl) propan-2- yl)amino)butan-2-ol dihydrochloride (7) (1.60 g, 3.16 mmol) solution in DCM (10 mL) and DIPEA (2.73 mL, 15.8 mmol) were added to the reaction mixture. The mixture was stirred at room temperature for 5 h and diluted with water. The organic phase was separated and the aqueous phase was washed with DCM (2 x 15 mL). Combined organic phase was washed with brine, and dried over Na₂SO₄. The solids were filtered off, and the solvent was removed by rotary evaporation. The crude product was purified by flash chromatography on silica gel eluting with EtOAc to give product 9a (1.05 g, 56%) as colourless oil. [053] ¹H NMR (300 MHz, Chloroform-d) δ 8.29 (d, J = 1.8 Hz, 1H), 8.14 (dt, J = 7.8, 1.4 Hz, 1H), 7.92 (dt, J = 7.8, 1.5 Hz, 1H), 7.48 (t, J = 7.8 Hz, 1H), 7.42 – 7.28 (m, 5H), 7.25 – 7.15 (m, 3H), 7.05 – 6.98 (m, 2H), 6.88 – 6.78 (m, 3H), 6.78 – 6.71 (m, 1H), 5.00 (s, 2H), 4.50 – 4.32 (m, 1H), 3.90 (s, 3H), 3.76 (s, 3H), 3.54 (dt, J = 6.9, 3.9 Hz, 1H), 3.02 (dd, J = 14.0, 7.1 Hz, 1H), 2.88 (dd, J = 14.0, 6.1 Hz, 1H), 2.59 – 2.41 (m, 2H), 1.52 (d, J = 2.1 Hz, 6H). HRMS [M+H]⁺: 597.2980, calcd.: 597.2965. [054] By a method analogous to Method A, the following compounds were obtained: Synthesis of intermediates 10, general method B Exemplified by the synthesis of methyl 3-(((2S,3R)-1-(3-(benzyloxy)phenyl)-3-((tert- butyldimethylsilyl)oxy)-4-((2-(3-methoxyphenyl)propan-2-yl)amino)butan-2- yl)carbamoyl)benzoate (10a). Methyl 3-(((2S,3R)-1-(3-(benzyloxy)phenyl)-3-hydroxy-4-((2-(3-methoxyphen-yl)propan-2- yl)amino)butan-2-yl)carbamoyl)benzoate (9a) (1.05 g, 1.76 mmol) was weighted in a glass vial and dissolved in DMF (15 mL). Imidazole (599 mg, 8.8 mmol) was added followed by TBDMSCl (1.32 g, 8.80 mmol). The reaction mixture was left stirring overnight heating at 80°C in oil bath (full conversion by UPLC). The reaction mixture was evaporated and the residue was separated between diethylether (20 mL) and water (20 mL). The organic phase was separated and the aqueous phase washed with diethylether (2 x 30 mL). Combined organic phase was washed with brine (20 mL), dried over Na₂SO₄, filtered and evaporated. The residue was purified by column chromatography on silica gel, eluting with 100% PE, then PE : EtOAc, 10:4. The product 10a (1.05 g, 84%) was collected as colorless oil. [055] ¹H NMR (300 MHz, Chloroform-d) δ 9.64 (d, J = 6.9 Hz, 1H), 8.51 (dt, J = 1.8, 1.0 Hz, 1H), 8.30 – 8.02 (m, 2H), 7.53 (t, J = 7.7 Hz, 1H), 7.44 – 7.27 (m, 6H), 7.25 – 7.13 (m, 1H), 7.01 – 6.93 (m, 2H), 6.85 – 6.74 (m, 4H), 5.02 (s, 2H), 4.54 – 4.40 (m, 1H), 3.89 (s, 3H), 3.74 (s, 3H), 3.67 - 3.61 (m, 1H), 3.20 (dd, J = 13.6, 5.2 Hz, 1H), 2.74 - 2.62 (m, 1H), 2.56 (dd, J = 11.7, 4.6 Hz, 1H), 2.40 (dd, J = 13.6, 10.2 Hz, 1H), 1.56 (s, 3H), 1.52 (s, 3H), 0.80 (s, 9H), -0.12 (s, 3H), -0.16 (s, 3H). HRMS [M+H]⁺: 711.3833, calc.: 711.3829 [056] By a method analogous to Method B, the following compounds were obtained: [057] Synthesis of intermediates 11, general method C Exemplified by the synthesis of methyl 3-(((2S,3R)-3-((tert-butyldimethylsilyl)oxy)-1-(3- hydroxyphenyl)-4-((2-(3-methoxyphenyl)propan-2-yl)amino)butan-2-yl)carbamoyl)-5- (dipropylcarbamoyl)benzoate (11a). [058] The solution of precursor 10a (500 mg, 0.70 mmol) in iPrOH (20 mL) was placed in Teflon cartridge, then Pd on C was added and the cartridge was placed in a hydrogenation reactor. Hydrogen (6 atm) was applied and the reaction mixture was stirred for 16 h (full conversion by UPLC). The reaction mixture was filtered through syringe filter (Sartorius RC 0.20 um), this was washed with MeOH and the filtrate was evaporated. The residue was dissolved in DCM and purified via column chromatography on silica gel eluting with DCM then DCM:MeOH, 50:1. Product 11a (390 mg, 89%) was obtained as a colourless oil. [059] ¹H NMR (400 MHz, Chloroform-d) δ 9.60 (d, J = 7.0 Hz, 1H), 8.49 – 8.47 (m, 1H), 8.18 (dt, J = 7.8, 1.4 Hz, 1H), 8.10 (ddd, J = 7.8, 1.9, 1.2 Hz, 1H), 7.53 (td, J = 7.8, 0.6 Hz, 1H), 7.28 – 7.23 (m, 1H), 7.09 (t, J = 7.8 Hz, 1H), 7.00 – 6.94 (m, 2H), 6.80 – 6.75 (m, 2H), 6.68 (ddt, J = 11.9, 7.8, 1.1 Hz, 2H), 4.49 (qd, J = 9.1, 7.4, 2.7 Hz, 1H), 3.89 (s, 2H), 3.75 (s, 3H), 3.70 (ddd, J = 4.7, 3.2, 1.4 Hz, 1H), 3.12 (dd, J = 13.8, 5.7 Hz, 1H), 2.71 (dd, J = 11.8, 1.5 Hz, 1H), 2.60 (ddd, J = 11.8, 4.6, 1.1 Hz, 1H), 2.41 (dd, J = 13.8, 9.7 Hz, 1H), 1.56 (s, 3H), 1.51 (s, 3H), 0.80 (s, 9H), -0.09 (s, 3H), -0.13 (s, 3H).HRMS [M+H]⁺: 621.3367, calc.: 621.3360. [060] By a method analogous to Method C, the following compounds were obtained: [061] Synthesis of intermediates 13, general method D Exemplified by the synthesis of methyl 3-(((2S,3R)-3-((tert-butyldimethylsilyl)oxy)-4-((2-(3- methoxyphenyl)propan-2-yl)amino)-1-(3-(3-((2-nitro-N- propylphenyl)sulfonamido)propoxy)phenyl)butan-2-yl)carbamoyl) benzoate (13.2) To the solution of methyl 3-(((2S,3R)-3-((tert-butyldimethylsilyl)oxy)-1-(3-hydroxyphenyl)- 4-((2-(3-methoxyphenyl)propan-2-yl)amino)butan-2-yl)carba-moyl)-5- (dipropylcarbamoyl)benzoate (11a) (75 mg, 0.12 mmol) and N-(3-hydroxypropyl)-2-nitro-N- propylbenzenesulfonamide (12b) (73 mg, 0.24 mmol) in DCM (2 mL) was added PPh₃ (63 mg, 0.24 mmol) under a nitrogen atmosphere. Then DEAD (28 uL, 0.18 mmol) was added dropwise and the reaction mixture was stirred for 2 h. The residue was dissolved in diethylether and purified by column chromatography on silicagel eluting with 100% diethylether. The product was repeatedly purified by reverse phase flash chromatography (C₁₈ silica, gradient 100% water to 100% MeCN from 2 min to 25 min; flow rate 20 mL/ min). Product 13.2 was collected as colourless solid (80 mg, 73%). [062] ¹H NMR (400 MHz, Chloroform-d) δ 9.63 (d, J = 6.9 Hz, 1H), 8.50 (t, J = 1.7 Hz, 1H), 8.18 (dt, J = 7.8, 1.4 Hz, 1H), 8.11 (dt, J = 7.9, 1.5 Hz, 1H), 8.02 – 7.98 (m, 1H), 7.64 – 7.49 (m, 4H), 7.26 (t, J = 7.9 Hz, 1H), 7.12 (t, J = 7.8 Hz, 1H), 7.02 – 6.93 (m, 2H), 6.76 (ddd, J = 13.0, 9.4, 3.4 Hz, 2H), 6.67 – 6.60 (m, 2H), 4.51 – 4.40 (m, 1H), 3.92 – 3.83 (m, 5H), 3.74 (s, 3H), 3.68 – 3.64 (m, 1H), 3.52 – 3.42 (m, 2H), 3.35 – 3.26 (m, 2H), 3.15 (dd, J = 13.8, 5.4 Hz, 1H), 2.71 (d, J = 11.8 Hz, 1H), 2.60 (dd, J = 12.2, 4.3 Hz, 1H), 2.39 (dd, J = 13.7, 10.1 Hz, 1H), 2.04 – 1.95 (m, 3H), 1.65 – 1.50 (m, 8H), 0.88 (t, J = 7.4 Hz, 3H), 0.79 (s, 9H), -0.12 (s, 3H), -0.16 (s, 3H). LCMS (ESI) m/z: 905.2 [M+H]⁺. [063] By a method analogous to Method D, the following compounds were obtained: [064] Synthesis of intermediates 14, general method E Exemplified by the synthesis of methyl 3-(((2S,3R)-3-((tert-butyldimethylsilyl)oxy)-4-((2-(3- methoxyphenyl)propan-2-yl)amino)-1-(3-(3-(propylamino)propoxy)phenyl)butan-2- yl)carbamoyl)benzoate (14.2) To a solution of methyl 3-(((2S,3R)-3-((tert-butyldimethylsilyl)oxy)-4-((2-(3- methoxyphenyl)propan-2-yl)amino)-1-(3-(3-((2-nitro-N-propylphenyl) sulfon- amido)propoxy)phenyl)butan-2-yl)carbamoyl)benzoate (13.2) (80 mg, 0.088 mmol) in MeCN (2 mL) was added K₂CO₃ (49 mg, 0.35 mmol) and thiophenol (18 uL, 0.18 mmol) and the mixture was stirred under Argon atmosphere for 2 h (solution was turning yellow). The reaction mixture was diluted with DCM (5 mL), washed with water (10 mL) and 0.1 M NaOH (10 mL). The combined aqueous phase was extracted with DCM (2 x15 mL). Combined organic phase was washed with brine, dried over Na₂SO₄, filtered and evaporated. The residue was purified by column chromatography on silica gel, eluting with DCM and DCM : MeOH, 50:1 to 10:1. Product 14.2 (45 mg, 71%) was obtained as colourless solid. [065] ¹H NMR (400 MHz, Chloroform-d) δ 9.57 (d, J = 7.0 Hz, 1H), 8.47 (t, J = 1.7 Hz, 1H), 8.16 (dt, J = 7.8, 1.4 Hz, 1H), 8.08 (dt, J = 7.8, 1.4 Hz, 1H), 7.51 (t, J = 7.8 Hz, 1H), 7.24 (t, J = 8.0 Hz, 1H), 7.13 (t, J = 7.9 Hz, 1H), 6.99 – 6.96 (m, 1H), 6.95 (t, J = 2.1 Hz, 1H), 6.76 (ddd, J = 8.2, 2.5, 0.9 Hz, 2H), 6.72 (t, J = 2.0 Hz, 1H), 6.68 (ddd, J = 8.2, 2.6, 0.9 Hz, 1H), 4.45 (dtd, J = 9.4, 6.0, 3.0 Hz, 1H), 4.01 (t, J = 5.8 Hz, 2H), 3.88 (s, 3H), 3.73 (s, 3H), 3.69 (d, J = 3.6 Hz, 1H), 3.16 – 3.03 (m, 3H), 2.90 – 2.79 (m, 2H), 2.71 (dd, J = 11.8, 1.4 Hz, 1H), 2.59 (dd, J = 11.5, 4.7 Hz, 1H), 2.42 (dd, J = 13.7, 9.6 Hz, 1H), 2.28 (dq, J = 12.1, 6.1 Hz, 2H), 1.83 (h, J = 7.5 Hz, 2H), 1.56 (s, 3H), 1.52 (s, 3H), 0.97 (t, J = 7.4 Hz, 3H), 0.78 (s, 10H), -0.12 (s, 3H), -0.16 (s, 3H). HRMS [M+H]⁺: 720.4420, calcd.: 720.4408. [066] By a method analogous to Method E, the following compounds were obtained:

[067] Synthesis of macrocyclic amides 15, general method F Exemplified by the synthesis of (S)-4-((R)-1-hydroxy-2-((2-(3-methoxyphenyl)propan-2- yl)amino)ethyl)-11-propyl-7-oxa-3,11-diaza-1,6(1,3)-dibenzenacyclododecaphane-2,12-dione (15.2) [068] Methyl 3-(((2S,3R)-3-((tert-butyldimethylsilyl)oxy)-4-((2-(3-methoxyphen-yl)- propan-2-yl)amino)-1-(3-(3-(propylamino)propoxy)phenyl)butan-2-yl)carbamoyl)benzoate (14.2) (45 mg, 0.063 mmol) was dissolved in a THF (1 mL), then 1 M NaOH (125 uL, 0.125 mmol) was added and the reaction mixture was stirred for 24 h at room temperature. The reaction mixture was evaporated and dissolved in DCM (6.5 mL). HBTU (24 mg, 0.062 mmol) was added and reaction mixture was stirred for 1 h. Reaction mixture was evaporated. The residue was suspended in DCM and filtered through short silica column eluting with DCM then DCM : MeOH, 50:1. Fractions with product were combined and evaporated. The residue was dissolved in THF (2 mL) and 1 M TBAF solution in THF (244 uL, 244 mmol) was added at room temperature under Argon atmosphere. The reaction mixture was stirred overnight and evaporated. The residue was purified by column chromatography on silica gel eluting with DCM : MeOH, 15:1 to 10:1. The product 15.2 was collected as colourless solid (9 mg, 25%). HRMS [M+H]⁺: 574.3285, calc.: 574.3281 [069] By a method analogous to Method F, the following compounds were obtained: [070] Physicochemical characterization of compounds 15

[071] Synthesis of methyl 3-(dipropylcarbamoyl)-5-iodobenzoate (17) 3-Iodo-5-(methoxycarbonyl)benzoic acid (16) (280 mg, 0.91 mmol) was dissolved in DCM (10 mL). HBTU (381 mg, 1.01 mmol) was added followed by di-n-propylamine (263 mL, 1.92 mmol). The reaction mixture was stirred overnight at room temperature. Then it was diluted with DCM (15 mL), washed with water, sat. NH₄Cl, brine, dried over Na₂SO₄ and filtered. Evaporation of the filtrate gave product 17 (320 mg, 90%) as colourless oil. [072] ¹H NMR (400 MHz, Chloroform-d) δ 8.39 (t, J = 1.6 Hz, 1H), 7.97 (t, J = 1.5 Hz, 1H), 7.88 (t, J = 1.6 Hz, 1H), 3.93 (s, 3H), 3.43 (d, J = 8.0 Hz, 2H), 3.13 (t, J = 7.5 Hz, 2H), 1.69 (dd, J = 14.5, 6.9 Hz, 2H), 1.54 (dd, J = 13.7, 6.2 Hz, 2H), 0.98 (t, J = 7.6 Hz, 3H), 0.77 (t, J = 7.2 Hz, 3H). HRMS [M+H]⁺: 390.0579, calcd.: 390.0566 [073] Synthesis of 3-(dipropylcarbamoyl)-5-(methoxycarbonyl)benzoic acid (8b) A dry 20 mL round bottom flask was charged with methyl 3-(dipropylcarbamoyl)-5- iodobenzoate (17) (300 mg, 077 mmol) and THF (20 mL) under argon and cooled to -20ºC. i- PrMgCl*LiCl complex solution in THF (1.3 M, 0.89 mL, 1.16 mmol) was slowly added and orange solution was stirred at -20ºC for 1 h. Dry ice (5 g) was placed in a 500 mL Erlenmeyer flask to generate CO₂ which was passed through the reaction mixture (CO₂ was dried using tube filled with CaCl₂) until all dry ice has evaporated. Reaction mixture was evaporated. 0.1 M HCl (20 mL) was added to the residue, and resulting suspension was extracted with DCM (3 x 30 mL). Combined organic phase was washed with brine, dried over Na₂SO₄, filtered and evaporated. The residue was purified by column chromatography on silica gel, eluting with DCM : MeOH, 15:1. Product 8b (124 mg, 52%) was obtained as colourless oil. [074] ¹H NMR (400 MHz, Chloroform-d) δ 8.75 (t, J = 1.6 Hz, 1H), 8.27 (p, J = 1.7 Hz, 2H), 3.97 (s, 3H), 3.52 – 3.44 (m, 2H), 3.15 (t, J = 7.0 Hz, 2H), 1.79 – 1.65 (m, 2H), 1.61 – 1.50 (m, 2H), 1.00 (t, J = 7.2 Hz, 3H), 0.75 (t, J = 5.7 Hz, 3H). [075] HRMS [M+H]⁺: 368.1507, calcd: 368.1498. [076] Synthesis of intermediates 19a-d and 21, general method G Exemplified by the synthesis of N-(4-hydroxybutyl)-2-nitrobenzene sulfonamide (19c) 4-Amino-1-butanol (18c) (0.84 mL, 9.0 mmol) was dissolved in DCM (10 mL) followed by the addition of Et₃N (1.88 mL, 13.5 mmol). This solution was added slowly to the cooled (0°C) solution of o-nitrobenzenesulfonyl chloride (1.0 g, 4.52 mmol) in DCM (10 mL). Reaction mixture was stirred at room temperature for 2 h and quenched with a 0.1 M HCl solution (20 mL). Phases were separated and the aqueous phase was extracted with DCM (3 x 15mL), The combined organic layers were washed with brine and dried over Na₂SO₄. Product 19c (1.23 g, quant) was obtained as yellowish oil. [077] ¹H NMR (400 MHz, Chloroform-d) δ 8.17 – 8.09 (m, 1H), 7.89 – 7.81 (m, 1H), 7.78 – 7.68 (m, 2H), 5.60 (t, J = 6.1 Hz, 1H), 3.65 (t, J = 5.8 Hz, 2H), 3.15 (q, J = 6.4 Hz, 2H), 1.70 – 1.56 (m, 4H). LCMS (ESI) m/z: 275.0 [M+H]⁺. [078] By a method analogous to Method G, the following compounds were obtained: [079] Synthesis of intermediates 12a-d, general method H Exemplified by the synthesis of N-(4-hydroxybutyl)-2-nitro-N-propylbenzene-sulfonamide (12c) N-(4-Hydroxybutyl)-2-nitrobenzenesulfonamide (19c) (1.23 g, 4.50 mmol) was dissolved in anhydrous DMF (20 mL) and bromopropane (1.23 mL, 13.5 mmol) was added, followed by K₂CO₃ (3.73 g, 27.0 mmol). The reaction mixture was stirred overnight at room temperature then quenched with water (30 mL) and extracted with DCM (3x30 mL). The combined organic phases were washed with brine and dried over Na₂SO_4. The solvent was evaporated and the residue was purified by column chromatography on silica gel, eluting with PE : EtOAc, 1:1 to give product 12c (1.32 g, 93%) as yellow oil. [080] ¹H NMR (400 MHz, Chloroform-d) δ 8.03 – 7.94 (m, 1H), 7.73 – 7.61 (m, 2H), 7.65 – 7.55 (m, 1H), 3.62 (t, J = 6.2 Hz, 2H), 3.36 – 3.27 (m, 2H), 3.28 – 3.19 (m, 2H), 1.68 – 1.46 (m, 6H), 0.83 (t, J = 7.4 Hz, 3H). LCMS (ESI) m/z: 317.1 [M+H]⁺. [081] By a method analogous to Method H, the following compounds were obtained: [082] Synthesis of intermediate 12e N-Neopentyl-2-nitrobenzenesulfonamide (21) (570 mg, 2.09 mmol) was dissolved in DMF (10 mL) under Argon flow. (4-bromobutoxy)(tert-butyl)dimethylsilane (1.12 g, 4.19 mmol) was added, followed by NaOH (167 mg, 4.19 mmol) and reaction mixture was stirred overnight at room temperature. Reaction mixture was quenched with water (20 mL) and extracted with diethylether (2 x 20 mL). The organic extracts were washed with brine, dried over Na₂SO₄, filtered and evaporated. The residue was dissolved in MeOH (40 mL). Ammonium fluoride (1.16 g, 31.4 mmol) was added and reaction mixture was stirred for 100 h at room temperature. Reaction mixture was charged with silica gel, MeOH was evaporated and the silica gel was dried. The material was poured onto silica gel column and eluted with PE:EtOAc, 10:4 to give product 12e (320 mg, 44%) as colourless oil. [083] ¹H NMR (400 MHz, Chloroform-d) δ 8.04 – 8.01 (m, 1H), 7.71 – 7.64 (m, 2H), 7.62 – 7.59 (m, 1H), 3.55 (t, J = 6.3 Hz, 2H), 3.41 – 3.35 (m, 2H), 3.18 (s, 2H), 1.64 – 1.54 (m, 2H), 1.45 – 1.35 (m, 2H), 0.96 (s, 9H). HRMS [M+H]⁺: 345.1484, calcd.: 345.1484 [084] In vitro Assay The compounds have been tested for antimalarials activity in vitro as malarial plasmepsin inhibitors according to the following process. [085] Determination of IC₅₀ Fluorescence resonance energy transfer (FRET) assay was performed to evaluate ability of compounds to inhibit plasmepsin IV and Cathepsin D. Solution of compounds for testing (concentration 0.01-100uM) on 96 well plate were added to the enzyme (Plasmepsin IV or Cathepsin D) in buffer (0.1 M NaOAc, pH = 4.5, 10% glycerin). The mixture was incubated for 30 min at 37ºC. Substrate (DABCYL-Glu-Arg-Nle-Phe-Leu-Ser-Phe-Pro-EDANS, AnaSpec Inc) was added to reach final concentration 5 uM. Hydrolysis of substrate was detected as an increase in fluorescence (Em 490 nm, Ex 336 nm) at 37ºC. Compounds were tested in three repeated triplicate experiments. [086] IC₅₀ values for selected inhibitors on plasmepsin IV (Plm IV) and cathepsin D (Plm IV) are given in Table 1 Table 1 Biological activity of macrocyclic amides 15

Claims

Claims 1. A compound selected from general formula I wherein: R¹, R², R³ are independently H, C_1-6alkyl, cycloC_3-12alkyl, cycloC_3-12alkyl-C_1-6alkyl, C_{2- 6}alkenyl, C_2-6alkynyl, aryl, biaryl, arylC_1-6alkyl, arylC_2-6alkenyl, arylC_2-6alkynyl, heteroaryl, heteroarylC_1-6alkyl, heteroarylC_2-6alkenyl, R⁴O(CH₂)_n, R⁴S(CH₂)_n, R⁴OC(=O)(CH₂)_n, R⁴N(R⁵)C(=O)(CH₂)_n, R⁴N(R⁵)(CH₂)_n, -F, -Cl, -Br, -I, -CF₃, -CH₂CF₃, -CF₂CF₂H, -OH, -L-OH,-O-L-OH, -OR⁴, -O-L-NH₂, -O-L-NHR⁴, -O-L-N(R⁴)R⁵, -L-OR⁴,-O-L-OR⁴,-OCF₃, -OCH₂CF₃, -OCF₂CF₂H, -L-OR⁴,-O-L-OR⁴,-OCF₃, -OCH₂CF₃, -OCF₂CF₂H, SR⁴, SCF₃, - CN, -NO₂, -NO₂, -NH₂, -NHR⁴, -NR⁴ ₂, -N(R⁴)R⁵, -L-NH₂, -L-NHR⁴, -L-NR⁴ ₂, -L-N(R⁴) R⁵,-NH-L-NH₂, -NH-L-NHR⁴, -NH-L-N(R⁴)R⁵, -NH-L-N(R⁴)R⁵,-NR⁴-L-NH₂, -NR⁴-L-NHR⁴, -NR⁴-L-N(R⁴)R⁵, -NR⁴-L-N(R⁴)R⁵, -N(R⁴)R⁵, -C(=O)OH, -C(=O)OR⁴, -C(=O)NH₂, -C(=O)NHR⁴, -C(=O)N(R ⁴)R⁵, -C(=O)N(R⁴)R⁵,-NHC(=O)R⁴, -NR⁴C(=O)R⁵, -NHC(=O)OR⁴, -NR⁴C(=O)OR⁵, -OC(=O)NH₂, -OC(=O)NHR⁴, -OC(=O)N(R⁴)R⁵, -OC(=O) R⁴NR⁵,-OC(=O)R⁴, -C(=O)R⁴,-NHC(=O)NH₂, -NHC(=O)NHR⁴, -NHC(=O)NR⁴ ₂, -NHC(=O)N(R⁴)R⁵, -NR⁴C(=O)NH₂, -N(R⁴)C(=O)NHR⁵, -NR⁴C(=O)N (R⁴)R⁵ wherein: n is an integer selected from 1 to 6; L represents -W^A-X^A-Y^A-Z^A-; or -W^A-X^A-Y^A-, or -W^A-X^A- or -W^A- wherein: W^A- represents a single bond, oxygen, sulfur, -NR⁶ or –CR⁶R⁷, X^A represents oxygen, sulfur, -NR⁶or –C(R⁶)R⁷, Y^A represents oxygen, sulfur, -NR⁶ or –C(R⁶)R⁷, Z^A represents oxygen, sulfur, -NR⁶ or –C(R⁶)R⁷; wherein: R⁶ and R⁷ are independently H, C_1-6alkyl, cycloC_3-12alkyl, cycloC_3-12alkyl-C_1-6alkyl, C_{2- 6}alkenyl, C_2-6alkynyl, aryl, biaryl, arylC_1-6alkyl, arylC_2-6alkenyl, arylC_2-6alkynyl, heteroaryl, heteroarylC_1-6alkyl, heteroarylC_2-6alkenyl, heteroarylthio, 2,3-dihydro-1H-indenyl, C_{1- 6}alkoxyC_1-6alkyl, aryloxyarylC_1-6alkoxy, C_1-6alkylthio, C_4-6alkenylthio, cycloC_3-12alkylthio, cycloC_3-12alkyl-C_1-6alkylthio, cycloC_3-12alkyl-C_3-6alkenylthio, C_1-6alkoxyC_1-6alkylthio, C_{1- 6}alkoxyC_3-6alkenylthio, arylC_3-6alkenylthio, heteroarylC_1-6alkylthio, C_1-6alkylsulfonyl, cycloC_3-12alkyl-C_1-6alkylsulfonyl, arylC_1-6alkylsulfonyl, C_1-6alkylamino, di-C_1-6alkylamino, cycloC_3-12alkylamino, C₁-C₆alkoxy-cycloC₃-C₁₂alkylamino, cycloC_3-12alkyl-C_1-6alkylamino, di-C_1-6alkylaminoC_1-6alkyl, C_1-6alkoxy-C_2-6alkylamino, arylamino, arylC_1-6alkylamino, N- cycloC_3-12alkyl-N-C_1-6alkylamino, N-aryl-N-C_1-6alkylamino, N-arylC_1-6alkyl-N-C_{1- 6}alkylamino, 2-indanylamino, tetrahydrofuryl, pyrrolidino, piperidino, 4-arylpiperidino, 4- heteroarylpiperidino, morpholino, piperazino, 4-C_1-6alkylpiperazino, 4-arylpiperazino, hexamethyleneimino, benzazepinyl, 1,3-dihydro-2H-isoindol-2-yl, heteroarylC_1-6alkoxy, heteroarylamino, or heteroarylC_1-6alkylamino; Q represents -A-D-E-G-H-J-; or -A=D-E-G-H-J-; or -A-D=E-G-H-J-; or -A-D-E=G- H-J-; or -A-D-E-G= H-J-; or -A-D-E-G-H=J-; or -D-E-G-H-J-; or -D=E-G-H-J-; or -D-E=G-H-J-; or -D-E-G=H-J-; or -D-E-G-H=J-; or -E-G-H-J-; or -E=G-H-J-; or -E-G=H-J-; or -E-G-H=J-; or -G-H-J-; or -G=H-J-; or -G-H=J-; or -H-J-; or -H=J-; or -J- wherein: A represents, oxygen, sulfur, -NR⁸ or –CR⁸R⁹ or =CR⁸, D represents oxygen, sulfur, -NR¹⁰ or –CR¹⁰R¹¹ or =CR¹⁰, E represents oxygen, sulfur, -NR¹² or –CR¹²R¹³ or =CR¹², G represents oxygen, sulfur, -NR¹⁴ or –CR¹⁴R¹⁵ or =CR¹⁴, H represents oxygen, sulfur, -NR¹⁶or –CR¹⁶R¹⁷ or =CR¹⁶, J represents oxygen, sulfur, -NR¹⁸ or –CR¹⁸R¹⁹ or =CR¹⁸, R⁸, R⁹, R¹⁰, R¹¹ _, R¹², R¹³, R¹⁴, R¹⁵ _, R¹⁶ _, R¹⁷, R¹⁸, R¹⁹ are independently H, C_1-6alkyl, cycloC_{3- 12}alkyl, cycloC_3-12alkyl-C_1-6alkyl, C_2-6alkenyl, C_2-6alkynyl, aryl, biaryl, arylC_1-6alkyl, arylC_{2- 6}alkenyl, arylC_2-6alkynyl, heteroaryl, heteroarylC_1-6alkyl, heteroarylC_2-6alkenyl, heteroarylthio, 2,3-dihydro-1H-indenyl, C_1-6alkoxyC_1-6alkyl, aryloxyarylC_1-6alkoxy, C_{1- 6}alkylthio, C_4-6alkenylthio, cycloC_3-12alkylthio, cycloC_3-12alkyl-C_1-6alkylthio, cycloC_3-12alkyl- C_3-6alkenylthio, C_1-6alkoxyC_1-6alkylthio, C_1-6alkoxyC_3-6alkenylthio, arylC_3-6alkenylthio, heteroarylC_1-6alkylthio, C_1-6alkylsulfonyl, cycloC_3-12alkyl-C_1-6alkylsulfonyl, arylC_{1- 6}alkylsulfonyl, C_1-6alkylamino, di-C_1-6alkylamino, cycloC_3-12alkylamino, C₁-C₆alkoxy- cycloC₃-C₁₂alkylamino, cycloC_3-12alkyl-C_1-6alkylamino, di-C_1-6alkylaminoC_1-6alkyl, C_{1- 6}alkoxy-C_2-6alkylamino, arylamino, arylC_1-6alkylamino, N-cycloC_3-12alkyl-N-C_1-6alkylamino, N-aryl-N-C_1-6alkylamino, N-arylC_1-6alkyl-N-C_1-6alkylamino, 2-indanylamino, tetrahydrofuryl, pyrrolidino, piperidino, 4-arylpiperidino, 4-heteroarylpiperidino, morpholino, piperazino, 4-C_1-6alkylpiperazino, 4-arylpiperazino, hexamethyleneimino, benzazepinyl, 1,3-dihydro-2H-isoindol-2-yl, heteroarylC_1-6alkoxy, heteroarylamino, or heteroarylC_{1- 6}alkylamino; or R⁸ and R⁹; R¹⁰ and R¹¹ _; R¹² and R¹³; R¹⁴, and R¹⁵ _; R¹⁶ and R¹⁷, R¹⁸ and R¹⁹ are independently =O,=NR²⁰ or =NOR²⁰; wherein: R²⁰ are independently H, C_1-6alkyl, cycloC_3-12alkyl, cycloC_3-12alkyl-C_1-6alkyl, C_2-6alkenyl, C_{2- 6}alkynyl, aryl, biaryl, arylC_1-6alkyl, arylC_2-6alkenyl, arylC_2-6alkynyl, heteroaryl, heteroarylC_1-6alkyl, heteroarylC_2-6alkenyl wherein: R⁸ and R⁹; R¹⁰ and R¹¹ _; R¹² and R¹³; R¹⁴, and R¹⁵ _; R¹⁶ and R¹⁷, R¹⁸ and R¹⁹ _; R⁸ and R¹⁰; R⁸ and R¹²; R⁸ and R¹⁴ _; R⁸ and R¹⁶; R⁸ and R¹⁸; R¹⁰ and R¹²; R¹⁰ and R¹⁴ _; R¹⁰ and R¹⁶; R¹⁰ and R¹⁸; R¹² and R¹⁴ _; R¹² and R¹⁶; R¹² and R¹⁸; R¹⁴ and R¹⁶; R¹⁴ and R¹⁸ R¹⁶ and R¹⁸ taken together represent -W^B-X^B-Y^B-Z^B-, or -W^B-X^B-Y^B-, or -W^B-X^B- or -W^B- wherein: W^B represents a single bond, oxygen, sulfur, -NR²¹ or –CR²¹R²², X^B represents oxygen, sulfur, -NR²¹ or –C(R²¹)R²², Y^B represents oxygen, sulfur, -NR²¹ or –C(R²¹)R²², Z^B represents oxygen, sulfur, -NR²¹ or –C(R²¹)R²², wherein: R²¹ and R²² are independently H, C_1-6alkyl, cycloC_3-12alkyl, cycloC_3-12alkyl-C_1-6alkyl, C_{2- 6}alkenyl, C_2-6alkynyl, aryl, biaryl, arylC_1-6alkyl, arylC_2-6alkenyl, arylC_2-6alkynyl, heteroaryl, heteroarylC_1-6alkyl, heteroarylC_2-6alkenyl, heteroarylthio, 2,3-dihydro-1H-indenyl, C_{1- 6}alkoxyC_1-6alkyl, aryloxyarylC_1-6alkoxy, C_1-6alkylthio, C_4-6alkenylthio, cycloC_3-12alkylthio, cycloC_3-12alkyl-C_1-6alkylthio, cycloC_3-12alkyl-C_3-6alkenylthio, C_1-6alkoxyC_1-6alkylthio, C_{1- 6}alkoxyC_3-6alkenylthio, arylC_3-6alkenylthio, heteroarylC_1-6alkylthio, C_1-6alkylsulfonyl, cycloC_3-12alkyl-C_1-6alkylsulfonyl, arylC_1-6alkylsulfonyl, C_1-6alkylamino, di-C_1-6alkylamino, cycloC_3-12alkylamino, C₁-C₆alkoxy-cycloC₃-C₁₂alkylamino, cycloC_3-12alkyl-C_1-6alkylamino, di-C_1-6alkylaminoC_1-6alkyl, C_1-6alkoxy-C_2-6alkylamino, arylamino, arylC_1-6alkylamino, N- cycloC_3-12alkyl-N-C_1-6alkylamino, N-aryl-N-C_1-6alkylamino, N-arylC_1-6alkyl-N-C_{1- 6}alkylamino, 2-indanylamino, tetrahydrofuryl, pyrrolidino, piperidino, 4-arylpiperidino, 4- heteroarylpiperidino, morpholino, piperazino, 4-C_1-6alkylpiperazino, 4-arylpiperazino, hexamethyleneimino, benzazepinyl, 1,3-dihydro-2H-isoindol-2-yl, heteroarylC_1-6alkoxy, heteroarylamino, or heteroarylC_1-6alkylamino; and optical isomers, pharmaceutically acceptable salts, hydrates, solvates, and polymorphs thereof. 2. The compound according to Claim 1 for use in the treatment of a disease that is mediated by aspartic proteases. 3. The compound according to Claim 2 wherein the disease is malaria. 4. A pharmaceutical composition comprising the compound according to Claim 1 and (i) macrocyclic amide or a pharmaceutically acceptable salt, solvate, morphological form and prodrug thereof, and (ii) a pharmaceutically acceptable carrier. 5. The pharmaceutical composition according to Claim 4, wherein said medicament is administered to human.