The present invention provides novel, conformationally constrained macrocyclic compounds of formula (I), as described herein below, having a substituent E that contains at least one ester moiety.

(I)

These conformationally constrained macrocyclic compounds exhibit antiproliferative activity on various cancer cell lines. In addition, they can be metabolized to compounds with modulating activity on the peptidyl-prolyl cis/trans isomerase Pin1 and may thus be useful in the treatment or prevention of a variety of diseases, conditions and disorders mediated by or sustained through the activity of Pin1 , or in the support of therapeutic treatments of specific disease conditions of primarily different cause. The present invention relates to methods of using these compounds in the treatment of various diseases and disorders, and to pharmaceutical compositions and forms comprising these compounds.

Isomerization of the peptidyl-prolyl bond plays an important role in many biological processes, including protein folding and regulation of various signaling pathways. Peptidyl-prolyl cis/trans isomerases (PPIases) are able to catalyse this conformational interconversion of peptidyl-prolyl bonds, and three classes of PPIases have been identified (S. D. Hanes, Biochim. Biophys. Acta 20 5, 1850(10), 2017- 2034 and literature cited therein).

The peptidyl-prolyl isomerase Pin1 (protein interacting with NIMA 1 (K. P. Lu et al., Nature 1996, 380(6574), 544-547)), belonging to the parvulin class of PPIase, is a phosphorylation-dependent peptidyl-prolyl isomerase that shows a unique substrate specificity for phosphorylated Ser/Thr-Pro motif(s). This conformational isomerization by Pin1 has been reported to be critically involved in diverse regulatory processes (K. P. Lu et a/., Nat. Chem Biol. 2007, 3(10), 619-629), affecting the function, protein- protein interactions, subcellular localization, protein phosphorylation, and stability of corresponding substrate proteins (Y.-C. Liou et al., Trends Biochem. Sci. 201 1 , 36(10), 501-514 and literature cited therein; K. P. Lu et al., Trends Cell. Biol. 2002, 12, 164-172). Human Pin1 is a small protein of 163 amino acids, comprising an N-terminal WW domain for substrate recognition, a flexible linker, and a C-terminal catalytic peptidyl- prolyl isomerase (PPI) domain (P.-J. Lu et al, J. Biol. Chem. 2002, 277(4), 2381 - 2384). High evolutionary conservation was reported for the Pin1 enzyme among eukaryotes, including mammalian Pin1 and orthologues in yeast (S. D. Hanes et al, Yeast 1989, 5(1), 55-72) and drosophila (T. Hsu et al, Nat. Cell Biol. 2001 , 3, 538- 543).

The crystal structure of human full-length Pin1 , complexed with the dipeptide Ala-Pro, was first reported by R. Ranganathan et al. (Ce// 1997, 89(6), 875-886). Further high- resolution crystal structures of Pin1 or mutants of Pin1 , in complex with peptidic or non-peptidic compounds, have since been published (Y. Zhang et al, ACS Chem. Biol. 2007, 2(5), 320-328; WO2004/005315 A2). Importantly, structural analysis revealed key binding interactions between Pin1 and the phosphorylated Ser/Thr-Pro motif in the substrates. These interactions include the formation of salt bridge(s) in a phosphate binding pocket and hydrophobic interactions in a proline binding pocket.

Rational design and various screening approaches have been used successfully for the discovery of Pin1 inhibitors, comprising peptidic and non-peptidic compounds. Only recently, Pin1 was identified as target of all- trans retinoic acid (S. Wei eta/., Nat. Med. 2015, 21 (5), 457-466) which is used in the therapy for acute promyelotic leukemia and was also applied in clinical trials for treatment of advanced breast cancer. Other known Pin1 inhibitors are less advanced. For many of these compounds, potential liabilities concerning selectivity and/or stability and/or potency and/or activity on whole cells were reported (J. D. Moore, A. Potter, Bioorg. Med. Chem. Lett. 2013, 23, 4283-4291 and literature cited therein). Among peptidic Pin1 inhibitors, linear pentameric peptides (D. Wildemann et a/., J. Med. Chem. 2006, 49(7), 2147-2150), cyclic heptameric peptides (T. Liu et a/., J. Med. Chem. 2010, 53(6), 2494-250), and disulfide-bridged nonameric peptides (K. E. Duncon et a , J. Med. Chem. 2011 , 54(11), 3854-3865) have been described. Furthermore, a series of linear peptides, including octameric peptides, were disclosed in WO2006/019982 A2 as Pin1 modulating compounds. As a general feature, peptidic Pin1 inhibitors comprise elements that mimic the phospho-Ser/Thr and/or proline moieties of the phosphorylated Ser/Thr-Pro motif in Pin1 substrates. Mimicry of the phospho-Ser/Thr moiety was mainly achieved by phosphonic or phosphoric acid-bearing residues, and corresponding peptidic inhibitors of Pin1 were frequently reported to be inactive or only weakly active in whole cell experiments. This issue was addressed by a combination strategy based on cell-penetrating peptides (W. Lian eta/., J. Am. Chem. Soc. 2014, 136(28), 9830-9833; T. Liu eta/., J. Med. Chem. 2010, 53(6), 2494-2501 ) or masking the acidic moiety by a prodrug-type ester (WO2006/124494 A1 ).

Several small molecule Pin1 inhibitors have been published in addition to the above- mentioned a\\-trans retinoic acid, including, naphthoquinone juglone, a non-reversible Pin1 inhibitor (L. Hennig et a/., Biochemistry 1998, 37, 5953-5960), hydroxy- naphthoquinone buparvaquone, used for treatment of certain parasitic infections in animals (J. Masolier et a/, Nature 2015, 520, 378-382), compounds comprising various acidic functional groups to mimic the phosphate group of the substrate (C. Guo et a/., Bioorg. Med. Chem. Lett. 2014, 24(17), 4187-4191 and literature cited therein), compounds sharing the presence of a carboxylic acid moiety (A. Potter et a/, Bioorg. Med. Chem. Lett. 2010, 20, 6483-6488), and a series of compounds all comprising an oxalic acid moiety (C. Liu et a/., Bioorg. Med. Chem. Lett. 2012, 20, 2992-2999). To improve cell-permeability masking of a phosphoric acid moiety as prodrug-type ester was described (S. Zhao, F. A. Etzkorn, Bioorg. Med. Chem. Lett. 2007, 17(23), 6615-6618). Furthermore, inhibitors of Pin1 have been disclosed in WO2004/087720 A1 , WO2006/040646 A1 , and WO2015/032998 A1 . The majority of these compounds share the presence of an acidic functional group, including carboxylic, phosphoric, phosphonic or sulfonic acids, and corresponding esters have also been disclosed, for example in WO2004/087720 A1 . In addition to the treatment of acute promyelotic leukemia, evidence is emerging that Pin1 modulators may be useful in the treatment or prevention of other diseases and conditions related to abnormal cell growth (Z. Lu, T. Hunter, Cell Res. 2014, 24, 1033-1049 and literature cited therein; E. S. Yeh, A. R. Means, Nat. Rev. Cancer 2007, 7, 381 -387). For example, Pin1 has been reported to control normal and cancer stem cells in the human breast (Rustighi A. et a!, Mol. Med. 2014, 6(1), 99- 1 19), including effects mediated through the p53 pathway (J. E. Giardini et a!, Cancer Cell 2011 , 12, 79-91 ; F. Mantovani et a/., Biochim. Biophys. Acta 2015, 1850(10), 2048-2060). Evidence on the role of Pin1 in tumorigenesis of breast cancer has also been obtained from studies in in vivo models (G. Wulf eta/., EMBO J. 2004, 23, 3397-3407).

Furthermore, overexpression of Pin1 has been reported in many cancers (L. Bao et a/, Am. J. Pathol. 2004, 164, M JA l'il), and the use of Pin1 inhibitors has been suggested for the treatment or prevention of diverse cancers, including breast cancer (J. Z. Wang et ai, Pharmacol. Res. 2015, 93, 28-35), prostate cancer (A. Ryo et ai, Clin. Cancer Res. 2005, 11(20), 7523-7531 ; S.-Y. Chen et ai, Mol. Cell. Biol. 2006, 26(3), 929-939), cervical cancer (H. Li et ai, Oncol Rep. 2006, 16, 491-496), liver cancer (R. W. Pang et ai, J. Pathol. 2006, 210, 19-25; G. Kim et ai, Biol. Pharm. Bull. 2015, 38(7), 975-979), lung cancer (X. Tan et ai, Cancer Biol. Ther. 2010, 9(2), 1 1 1 -1 19), esophageal cancer (H. Jin etai, Oncol. Lett. 201 1 , 2(6), 1 191-1 196), colon cancer (C. J. Wim et ai, World J. Gastroenterol. 2005, 11(32), 5006-5009), gastric cancer (M. Shi et ai, Cell Biochem. Biophys. 2015, 71(2), 857-864), and lymphoma, such as non-Hodgkin lymphoma (G. Fan et ai, Cancer Res. 2009, 69(11), 4589- 4597).

From other studies it was also suggested that Pin1 inhibitors may be useful for the treatment and/or prevention of other diseases or conditions, including asthma (P. Anders, Nat. Immunol. 2005, 6, 121 1-1212), allergic pulmonary eosinophilia (S. Esnaut etai, J. Allergy Clin. Immunol. 2007, 120, 1082-1088), pulmonary fibrosis, for example caused by chronic asthma (Z.-J. Shen et ai, J. Clin. Invest. 2008, 118(2), 479-490), stroke (S. H. Baik et ai, Ann. Neurol. 2015, 77(3), 504-517), viral infections, for example HIV/AIDS (H. Hou et ai, Gene 20 5, 656 \ ), 9-14), infections with intracellular and extracellular pathogens, for example infections by intracellular pathogen Chlamydia trachomatis (A J. Olive et ai, Cell Host Microb. 2014, 15(1), 1 13-124), osteolytic bone diseases, for example periodontitis (Y.-A. Cho et ai, J. Dent. Res. 2015, 94(2), 371-380), cardiovascular diseases, for example diabetic vascular disease (F. Paneni et ai, Eur. Heart J. 2015, 36(13), 817-828), diabetic restenosis (L. Lv et ai, J. Cell Mol. Med. 2013, 17(8), 989-1005), nonalcoholic steatohepatitis (Y. Nakatsu et al., J. Biol. Chem. 2012, 287(53), 44526-44535), and cardiac hypertrophy (H. Toko et al., Circ. Res. 2013, 112(9), 1244-1252; S. Sakai et al, Life Sci. 2014, 102(2), 98-104). Results from recent studies indicate that Pin1 may be implicated in additional inflammatory diseases (T. Boussetta et al, Blood 2010, 116(25), 5795-5802), including rheumatoid arthritis, inflammatory bowel diseases, and acute respiratory distress syndrome. Immunosuppressive effects of a Pin1 inhibitor were reported from animal studies of organ transplantation (S. Esnault et al, PLoS One 2007, 2, e226). Accordingly, important roles for Pin1 were also suggested in immune disorders like diabetes, multiple sclerosis and lupus, macrophage mediated tissue damage, gastritis, and myeloproliferative syndromes (Z.-H. Shen, J. S. Malter, Biomoiecuies 2015, 5, 412-434). In addition to cell-cycle-regulated proteins and transcription factors, Alzheimer's disease-related proteins have been identified as substrates of Pin1 . Recent studies support a neuroprotective role of Pin1 with opposite directions of Pin1 dysregulation in Alzheimer's disease as compared to cancer (J. A. Driver et al, Biochim Biphys. Acta 2M5, 1850(10), 2069-2076; J. A. Driver et al., Discov. Med. 2014, 17(92), 93- 99). Pin1 is also suggested to be involved in other neurodegenerative diseases, including Huntington's disease (M. T. Lin, M. F. Beal, Nature 2006, 443, 787-795; A. Grison et al., /VAS 2011 , 108(44), 17979-17984) and frontotemporal dementia that is associated with a mutation in the tau gene (J. Lim et al, J. Clin. Invest. 2008, 118(5), 1877-1889).

Pin1 inhibitors may also be useful for the treatment or prevention of certain parasite infections in animals, including infections of cattle with Theileria parasites (J. Masolier et al., Nature 2015, 520, 378-382). Importantly, the homologue of Pin1 in Theileria annulata was demonstrated to play a key role in maintaining bovine leukocyte transformation, and Pin1 inhibitor buparvaquone was able to reverse transformed phenotypes.

The compounds of the invention show antiproliferative activity on various cancer cell lines, including THP-1 , MHHES1 , A673, and BT20 cells. The THP-1 cell line has been used for the study of anticancer compounds, including antileukemic drugs (P. W. Hollenbach et al., PLoS CWE2010, 5(2); doi:10.1371/journal.pone.0009001 ), and has been reported as model for immune modulation studies (W. Chanput et ai, Int. Immunopharmacol. 2014, 23(1), 37-45). Furthermore, results from experiments using THP-1 cells suggested a role of Pin1 in nonalcoholic steatohepatitis (Y. Nakatsu et al., J. Biol. Chem. 2012, 287(53), 44526-44535) and in various immune diseases (A. Tun-Kyi eta/., Nat. Immunol. 2011 , 12(8), 733-741 ). The breast cancer cell line BT20, and Ewing's sarcoma cell lines MHHES1 and A673 have been applied in cancer research, including studies based on BT20 cells (A. D. Hughes et ai, Cancer Letters 2014, 352, 28-35; A. A. Pritsa et ai, Anticancer Drugs 2001 , 12(2), 137-142), studies based on MHHES1 cells (D. G. Gimenez et ai, Planta Med. 2010, 76(2), 133-136), and studies based on A673 cells (S. R. Ambati et ai, Mol. Oncol. 2014, 8(2), 323- 336; C. L. Cubitt etai, Sarcoma 2013; doi: 10.1 155/2013/365723).

The present invention provides new chemical entities which contain a macrocyclic backbone with appended substituents, including E, G, and Q. Substituent E comprises at least one ester moiety, that can be metabolized to liberate the corresponding acid moiety, which is involved in the mimicry of the phospho-Ser/Thr moiety of Pin1 substrates. Additionally, it is essential that compounds of the invention comprise an aromatic group in substituents G and/or Q, as described herein below. Compounds based on macrocyclic scaffolds and modular approaches for their synthesis have been described in the literature and also in patent applications WO201 1/014973 A2, WO201 1/015241 A1 and WO2013/139697 A1 . The three latter publications contain versatile methods to generate macrocyclic compounds using combinatorial and parallel synthesis strategies.

In a first embodiment (1), the present invention relates to compounds of formula (I)

(I)

wherein

L is -C(O)-; or -S(0) ₂ -;

Xis 0;S;-S(0)-;or-S(0) ₂ -;

V is -OC(O)-; -NR ⁷ C(0)-; -SC(O)-; -OS(0) ₂ -; or -NR ⁷ S(0) ₂ -; r is an integer of 1-2;

t is an integer of 0-1;

i and p are independently an integer of 0-3 with the proviso that 1≤ i+p≤ 3;

R ¹ , R ² , R ³ , R ⁴ , R ⁵ , and R ⁶ are independently H; F; or CH ₃ ;

with the proviso that

at most three substituents of R ¹ , R ² , R ³ , R ⁴ , R ⁵ , and R ⁶ are F; or CH ₃ ; R ⁷ is H; Ci-3 alkyl; or cyclopropyl;

R ⁸ is H; or F;

R ⁹ is H; F; CF ₃ ; orCi-3-alkyl;

R ¹⁰ , R ¹¹ , R ¹² , R ³ , and R ¹⁴ are independently H; F; or C1-3 alkyl;

with the proviso that

at most three substituents of R ¹⁰ , R ¹¹ , R ¹² , R ¹³ , and R ¹⁴ are F; or Ci _-3 alkyl; R ¹⁵ is H; or Ci- ₃ alkyl;

R ⁶ is H; F;orCH ₃ ; E is a group of one of the formulae

R ¹⁷ is -C(0)OR ^18a ; or -P(0)(OR ^18a )(OR ^18b );

R ^18a is Ci-4-alkyl; C3-4-cycloalkyl; benzyl; phenyl; or a group of formula

R ^8b is F1 ;

R ¹⁹ is Ci-4-alkyl; -CHR ²⁰ R ²¹ ; oxetan-2-yl; oxetan-3-yl; or -(CH ₂ )jCH ₂ OH;

R ²⁰ and R ²¹ are independently H; or Ci-4-alkyl;

R ²⁰ and R ²¹ together with the carbon atom to which they are connected can form C3-6-cycloalkyl moieties;

R ²² is H; Ci-2-alkyl; -(CHR ²³ ) ₀ C(0)OR ^8a ; -(CHR ²³ ) ₀ P(0)(OR ^8a )(OR ^8b );

-(CHR ²³ )oOH; -(CHR ²³ ) ₀ C(0)NH ₂ ; -(CHR ²³ ) ₀ NHC(0)NH ₂ ;

-(CHR ²³ )oS(0) ₂ NH ₂ ; or -(CHR ²³ ) ₀ OC(0)NH ₂ ;

R ²³ is H; F; or CH ₃ ;

R ^24a and R ^24b are independently H; or CH ₃ ;

Z is -C(O)-; -S(0) ₂ -; -OC(O)-; -OS(0) ₂ -; -NR ²⁴ C(0)-; or -NR ²⁴ S(0) ₂ -; Z ² is -C(O)-; -S(0) ₂ -; -(CHR ²³ )-; -OC(O)-; -OS(0) ₂ -; -NR ² C(0)-;

-NR ^{2 b} S(0) ₂ -; -C(0)NR ^{2 b} -; or -S(0) ₂ NR ^{2 b} -; d is an integer of 0-1 ;

j is an integer of 0-3;

if E is E1 , o is an integer of 1 -3; if E is E2, o is an integer of 0-2;

if E is E3; E4; E5; or E6; o is an integer of 0-1 ; with the proviso that

in E at most three substituents R ²³ are different from H;

G is H; Ci-6-alkyl; C2-6-alkenyl; C3-6-cycloalkyl; C3-6-heterocyclyl; C6-io-aryl;

C5-io-heteroaryl; or a group of one of the formulae

R ²⁵ is H; F; CH ₃ ; CF ₃ ; OCH ₃ ; OCF ₃ ; or OCHF ₂ ;

R ²⁶ is H; F; CI; CF ₃ ; OCF ₃ ; OCHF ₂ ; Ci _-2 -alkyl; Ci _-2 -alkoxy; or Ci _-2 -thioalkoxy; R ²⁷ is H; F; CI; CF ₃ ; OCF ₃ ; OCHF ₂ ; CN; Ci _-3 -alkyl; Ci _-3 -alkoxy;

Ci- ₃ -thioalkoxy; -C(0)NR ³ R ³² ; or -S(0) ₂ NR ³ R ³² ;

R ²⁸ is H; F; CI; CF ₃ ; CN; OCF ₃ ; OCHF ₂ ; Ci _-3 -alkyl; Ci _-3 -alkoxy; or

Ci _-3 -thioalkoxy;

R ²⁹ is H; F; CF ₃ ; OCF ₃ ; OCHF ₂ ; Ci _-3 -alkyl; Ci _-3 -alkoxy; or Ci _-3 -thioalkoxy; R ³⁰ is H; F; CI; CF ₃ ; OH; OCF ₃ ; OCHF ₂ ; N0 ₂ ; Ci _-3 -alkyl; Ci _-3 -alkoxy;

Ci- ₃ -thioalkoxy; or -NR ³ R ³² ;

R ³¹ and R ³² are independently H; or CH ₃ ;

R ³³ is H; Ci-3-alkyl; or -C(0)-Ci _-3 -alkyl;

T is N; or CR ²⁵ ;

M is 0; S; or NR ³³ ; with the proviso that

in each of G1 to G4 at least two of the substituents are H;

in each G5 to G10 at least one of the substituents is H; and with the proviso that,

if Q is O; S; -S(O)-; or -S(0) ₂ -; or

if R ³⁴ in Q1 is H ; F; CF ₃ ; OH ; SH ; Ci _-8 -alkyl; C ₂ - ₈ -alkenyl; C _2-8 -alkynyl; Ci-8-alkoxy; Ci-e-thioalkoxy; C _3- 8-cycloalkyl; C _3- 8-heterocyclyl; or

-N R ⁴⁸ C(0)C(0)OR ^18a ;

then G is C6-io-aryl; Cs-io-heteroaryl; or a group of one of the formulae G1 to G14;

Q is O; S; -S(O)-; -S(0) ₂ -; or a group of one of the formulae

Z ³ is -(CHR ⁴⁷ )-; O; -C(O)-; -C(0)NR ⁴⁸ -; -NR ⁴⁸ C(0)-; -NR ⁴⁸ C(0)NR ⁴⁸ -;

-NR ⁸ C(0)0-; -OC(0)NR ⁴⁸ -; -NR ⁸ S(0) ₂ -; -S(0) ₂ NR ⁴⁸ -; or

-NR ⁴⁸ S(0) ₂ NR ⁴⁸ -;

R ³⁴ is H; F; CF ₃ ; OH; SH; Ci _-8 -alkyl; C _2-8 -alkenyl; C _2-8 -alkynyl; Ci _-8 -alkoxy;

Ci-e-thioalkoxy; C _3- 8-cycloalkyl; C _3- 8-heterocyclyl; C6-io-aryl;

Cs-io-heteroaryl; or -NR ⁸ C(0)C(0)OR ^8a ;

R ³⁵ is H; or CH ₃ ;

R ³⁶ is a group of one of the formulae

R ³⁷ is H; F; CI; CH ₃ ; OCH ₃ ; CF ₃ ; OCF ₃ ; or OCHF ₂ ;

R ³⁸ is H; F; CI; CF ₃ ; OCF ₃ ; OCHF ₂ ; Ci-4-alkyl; C _2-4 -alkenyl; Ci _-4 -alkoxy;

Ci _-4 -thioalkoxy; or C _3-4 -cycloalkyl;

R ³⁹ is H; F; CI; Br; I; CF ₃ ; OH; OCF ₃ ; OCHF ₂ ; N0 ₂ ; CN; Ci _-6 -alkyl; Ci _-6 -alkoxy;

Ci-6-thioalkoxy; C _3-6 -cycloalkyl; C _3-6 -heterocyclyl; -C(0)OR ^18a ;

-C(O)NR ⁴⁰ R ⁴¹ ; or -S(O) ₂ NR ⁴⁰ R ⁴¹ ;

R ⁴⁰ and R ⁴¹ are independently H; or Ci _-3 -alkyl;

R ⁴² is C6-aryl-Ci- ₄ -alkyl; C5-6-heteroaryl-Ci _-4 -alkyl; or a group of formula

H11

R ⁴³ is C6-aryl; Cs-6-heteroaryl; or a group of one of the formulae

R is H; F; CI; CF ₃ ; OH; OCF ₃ ; OCHF ₂ ; N0 ₂ ; CN; Ci-4-alkyl; Ci-4-alkoxy;

Ci _-4 -thioalkoxy; or C _3-4 -cycloalkyl; R ⁴⁵ is H; F; CI; CF ₃ ; OCF ₃ ; OCHF ₂ ; Ci-4-alkyl; Ci-4-alkoxy; or Ci _-4 -thioalkoxy;

R ⁴⁶ is H; Ci-e-alkyl; -C(0)-Ci _-6 -alkyl; or -C(0)-C ₃ - ₆ -cycloalkyl;

R ⁷ is H; F; CI; CH ₃ ; or CF ₃ ;

R ⁸ is H; or Ci _-3 alkyl;

M' is O; S; or NR ⁴⁶ ;

T' is N; or CR ³⁷ ;

U is 0; S; or CHR ⁴⁷ ; m and n are independently an integer of 0-4 with the proviso that n+m<4;

e is an integer of 0-1 ;

g is an integer of 0-2; with the proviso that,

if G is H; Ci-6-alkyl; C2-6-alkenyl; C _3- 6-cycloalkyl; or C _3- 6-heterocyclyl;

then Q is Q1 ; or Q2; R ³⁴ in Q1 is C6-io-aryl; or Cs-io-heteroaryl; and with the proviso that,

if i is 1 ; or p is 0;

then Q is Q1 ; or Q2;

and with the further proviso that,

if G is H; C1-6 alkyl; C2-6-alkenyl; C _3- 6-cycloalkyl; or C _3- 6-heterocyclyl;

then R ³⁴ in Q1 is C6-io-aryl; or Cs-io-heteroaryl; and with the proviso that

in Q at most 3 substituents R ⁴⁷ are different from H; an aromatic 5-membered (t=0) or 6-membered (t=1 ) ring system is positioned between functional moieties L and X; if t=0,

Y is CR ⁴⁹ ; NR ⁵⁵ ; N; O; or S;

Y ² is CR ⁵⁰ ; NR ⁵⁵ ; N; O; or S;

Y ³ is CR ⁵¹ ; NR ⁵⁵ ; N; O; or S; with the proviso that

the aromatic 5-membered ring system is a group of one of the formulae

wherein Y' is NR ⁵⁵ ; O; or S; if t=1,

Y ¹ is CR ⁴⁹ ; or N

Y ² is CR ⁵⁰ ; or N

Y ³ is CR ⁵¹ ; orN

Y ⁴ is CR ⁵² ; or N with the proviso that

the aromatic 6-membered rin s stem is a rou of one of the formulae

^49

H32 and with the further proviso that

in H24 two of the substituents are H;

in each of H25 to H28 one of the substituents is H; R ⁴⁹ and R ⁵⁰ are independently H; F; CI; CH ₃ ; CH ₂ CH ₃ ; CF ₃ ; OCH ₃ ; OCF ₃ ; or

OCHF ₂ ;

R ⁵¹ is H; F; CI; CF ₃ ; OCF ₃ ; OCHF ₂ ; N0 ₂ ; NH ₂ ; OH; CN; Ci-4-alkyl; C _2-4 -alkenyl;

C _2- 4-alkynyl; C _3- 4-cycloalkyl; Ci-4-alkoxy; Ci-4-thioalkoxy; or -NR ⁵³ R ⁵⁴ ; R ⁵² is H; F; CI; CH ₃ ; CF ₃ ; OCF ₃ ; OCHF ₂ ; or OCH ₃ ;

R ⁵³ and R ⁵⁴ are independently H; or Ci _-2 -alkyl;

R ⁵³ and R ⁵⁴ together with the nitrogen atom to which they are connected can form C _3- 5-heterocyclyl moieties;

R ⁵⁵ is H; or CH ₃ ; and wherein

in each such compound at most 12 halogen substituents are present; or stereoisomers; or tautomers or rotamers thereof; or a salts; or a pharmaceutically acceptable salts; or a solvates thereof.

A bond drawn as dotted line indicates the point of attachment of the corresponding radical or substituent. For example, the drawing below

represents the 5-hydroxy-naphth-2-yl substituent.

For the avoidance of doubt, some of the aforementioned substituents, for example, but not limited to, R ³ , R ⁴ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ , R ^8a , R ^8b , R ¹⁹ , R ²⁰ , R ²¹ , R ²³ , R ⁴⁷ , and R ⁴⁸ ; as well as some of the indices (for example o, and g) may occur several times within the same molecular entity. In such a case each of them shall be selected independently from others specified by the same symbol, unless otherwise indicated.

"Salts" as understood herein are especially, but not limited to, the pharmaceutically acceptable salts of compounds of formula (I). Such salts are formed, for example, as acid addition salts with organic or inorganic acids, from macrocyclic compounds of the invention with a basic nitrogen atom. Suitable inorganic acids are, for example, halogen acids, such as hydrochloric acid, sulfuric acid, or phosphoric acid. Suitable organic acids are, for example, carboxylic, phosphonic, sulfonic or sulfamic acids; like acetic acid, propionic acid, octanoic acid, decanoic acid, dodecanoic acid, glycolic acid, lactic acid, fumaric acid, succinic acid, adipic acid, pimelic acid, suberic acid, azelaic acid, malic acid, tartaric acid, citric acid, amino acids, such as glutamic acid or aspartic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, cyclohexanecarboxylic acid, adamantine-carboxylic acid, benzoic acid, salicylic acid, 4 aminosalicylic acid, phthalic acid, phenylacetic acid, mandelic acid, cinnamic acid, methane- or ethane-sulfonic acid, 2-hydroxy-ethanesulfonic acid, ethane-1 ,2- disulfonic acid, benzene-sulfonic acid, 2-naphthalenesulfonic acid, 1 ,5-naphthalene- disulfonic acid, 2-, 3- or 4-methyl-benzene-sulfonic acid, methylsulfuric acid, ethyl- sulfuric acid, dodecylsulfuric acid, N-cyclohexylsulfamic acid, N-methyl-, N-ethyl- or N propyl-sulfamic acid, or other organic protonic acids, such as ascorbic acid.

As used in this description, the term "alkyl", taken alone or in combinations (i.e. as part of another group, such as "aryl-Ci-6-alkyl"), designates saturated, straight-chain or branched hydrocarbon radicals and, unless otherwise indicated, may be optionally substituted with at most 2 substituents selected from the group of F and CI. The term "Cx-y-alkyl" (x and y each being an integer) refers to an alkyl group as defined above containing x to y carbon atoms. For example a Ci-6-alkyl group contains one to six carbon atoms. Representative examples of alkyl groups include methyl, ethyl, n- propyl, /so-propyl, /7-butyl, /so-butyl, sec-butyl, fe -butyl, /7-pentyl, /7-hexyl and the like.

The term "alkenyl", taken alone or in combinations, designates straight chain or branched hydrocarbon radicals containing at least one or, depending on the chain length, up to four olefinic double bonds. Such alkenyl moieties, unless otherwise indicated, may be optionally substituted with at most 2 substituents selected from the group of F and CI, and can independently exist as E or Z configurations per double bond, which are all part of the invention. The term "C _x - _y -alkenyl" (x and y each being an integer) refers to an alkenyl group as defined above, containing x to y carbon atoms. Examples of this moiety include, but are not limited to, vinyl, prop-1-en-1-yl, 2- methylprop-1 -en-1 -yl, and allyl.

The term "alkynyl" designates straight chain or branched hydrocarbon radicals containing at least one or, depending on the chain length, up to four triple bonds. The term "C _x - _y -alkynyl" (x and y each being an integer) refers to an alkynyl group as defined above, containing x to y carbon atoms. Examples of this moiety include, but are not limited to, prop-2-yn-1 -yl.

The term "cycloalkyi" refers to a saturated or partially unsaturated alicyclic moiety having from three to eight carbon atoms and, unless otherwise indicated, may be optionally substituted with at most 2 substituents selected from the group of F and CI. The term "C _x-y -cycloalkyl" (x and y each being an integer) refers to a cycloalkyi group as defined above, containing x to y carbon atoms. Examples of this moiety include, but are not limited to, cyclobutyl, cyclohexyl, norbornyl and the like.

The term "heterocyclyl" describes a saturated or partially unsaturated mono- or bicyclic moiety having from one to seven ring carbon atoms and one or more ring heteroatoms selected from oxygen, sulphur or nitrogen, provided the nitrogen is forming an aromatic amino group, or is part of an amide, urea, urethane, or sulfonamide group within the heterocyclyl moiety. The term "C _x - _y -heterocyclyl" (x and y each being an integer) refers to a heterocyclyl group as defined above, containing x to y ring atoms. Examples of this moiety include, but are not limited to, morpholino, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, and the like.

The term "aryl", taken alone or in combinations, designates aromatic carbocyclic hydrocarbon radicals containing one or two six-membered rings. The term "C6-aryl" refers to phenyl. The term "C6-io-aryl" refers to phenyl or naphthyl, which, unless otherwise indicated, may be optionally substituted with at most 3 substituents selected from the group of F, CI, CF3, OCF3, and OCF2.

The term "heteroaryl", taken alone or in combinations, designates aromatic heterocyclic radicals containing one or two five- and/or six-membered rings, at least one of them containing up to four heteroatoms selected from the group consisting of O, S and N and whereby the heteroaryl radicals or tautomeric forms thereof may be attached via any suitable atom. The term "C _x-y -heteroaryl" (x and y each being an integer) refers to a heteraryl group as defined above, containing x to y ring atoms. Said heteroaryl ring(s) are optionally substituted, e.g. as indicated above for "aryl". Examples of the term "C5-6-heteroaryl" include, but are not limited to, furanyl, oxazolyl, isoxazolyl, thiophenyl, thiazolyl, isothiazolyl, pyrrolyl, pyrimidinyl, pyridyl and the like. Examples of the term "Cs-io-heteroaryl" include, but are not limited to, furanyl, oxazolyl, isoxazolyl, thiophenyl, thiazolyl, isothiazolyl, pyrrolyl, pyrimidinyl, pyridyl, quinolinyl, benzothiofuranyl and the like.

The term "-C(0)-C _x - _y -alkyl", as used herein, refers to an C _x-y -alkyl group as defined above, connected to a carbonyl group. Representative examples of -C(0)-C _x-y -alkyl moieties include, but are not limited to, acetyl, propanoyl, /so-butanoyl and the like. The term "-C(0)-C3-6-cycloalkyl", as used herein, refers to an C _x-y -cycloalkyl group as defined above, connected to a carbonyl group. Representative examples of -C(0)-C3-6-cycloalkyl moieties include, but are not limited to, cyclopropyl-methanoyl, cyclobutyl-methanoyl and the like. The term "C6-aryl-C _x-y -alkyl", as used herein, refers to an C _x-y -alkyl group as defined above, substituted by an C6-aryl group, as defined above. Representative examples of C6-aryl-C _x-y -alkyl moieties include, but are not limited to, benzyl, 1-phenylethyl, 2- phenylethyl, 3-phenylpropyl, 2-phenylpropyl and the like. The term "C5-6-heteroaryl-C _x-y -alkyl", as used herein, refers to an C _x-y -alkyl group as defined above, substituted by a Cs-6-heteroaryl group, as defined above. Examples of C5-6-heteroaryl-C _x-y -alkyl groups, for example, include pyridin-3-ylmethyl, (1 H-pyrrol-2- yl)ethyl and the like. The terms "alkoxy" and "aryloxy", taken alone or in combinations, refer to the groups of -O-alkyl and -O-aryl respectively, wherein an alkyl group or an aryl group is as defined above. The term "C _x-y -alkoxy" (x and y each being an integer) refers to an -O- alkyl group as defined above containing x to y carbon atoms attached to an oxygen atom. Representative examples of alkoxy groups include methoxy, ethoxy, n- propoxy, /so-propoxy, /7-butoxy, fe -butoxy and the like. Examples of aryloxy include e.g. phenoxy. The term "thioalkoxy", taken alone or in combinations, refers to an -S-alkyl group, wherein an alkyl group is as defined above. The term "C _x-y -thioalkoxy" (x and y each being an integer) refers to an -S-alkyl group as defined above containing x to y carbon atoms attached to an sulfur atom. Representative examples of thioalkoxy groups include methlythio, ethylthio and the like.

The term "halogen" refers to a fluorine substituent (F), a chlorine substituent (CI), a bromine substituent (Br) or an iodine substituent (I). "Amino" designates primary, secondary or tertiary amine groups. Particular secondary and tertiary amine groups are alkylamine, dialkylamine, arylamine, diarylamine, arylalkylamine and diarylamine groups wherein the alkyl or aryl is as herein defined and optionally substituted. For the avoidance of doubt the term "heteroatom" refers to any atom that is not carbon or hydrogen.

The term "isomer" comprises species of identical chemical formula, constitution and thus molecular mass, such as but not limited to C=C-double bond or amide cis/trans isomers, rotamers, conformers and diastereomers.

All possible stereoisomers - explicitly including atropisomers - conformers and rotamers as well as salts, solvates, clathrates, N-oxides, or isotopically enriched or enantiomerically enriched versions of macrocyclic compounds of formula (I) are part of this invention.

A further embodiment (2) of the invention relates to compounds of formula (I) according to embodiment (1 ),

wherein

3 stereocenters are further defined as in formula (II)

if Q is Q1 ;

then, in the moiety comprising Q1 , one further stereocenter is defined formula (III)

if Q is Q2;

then, in the moiety comprising Q2, one further stereocenter is defined as in formula (IV)

or tautomers or rotamers thereof; or salts; or pharmaceutically acceptable salts; or solvates thereof. A further embodiment (3) of the invention relates to compounds of formula (I) according to embodiment (2),

wherein

L is -C(O)-;

X is O;

V is -OC(O)-; r is 1;

tis 1;

i is 1, and p is 1; or

i is 2, and p is 1;

R ¹ to R ¹⁶ , and R ³⁵ are H; Eis E1;E2;orE3;

R ^18a is Ci-3-alkyl; orF1;

R ¹⁹ is Ci-4-alkyl;

R ²⁰ is H;

R ²¹ is H; orCi-3-alkyl;

R22 is H; -(CHR ²³ ) ₀ C(0)OR ^18a ; or -(CHR ²³ ) ₀ P(0)(OR ^18a )(OR ^18b );

R ²³ , R24a, and R ^24b are H;

Z is -C(O)-;

Z ² is -C(0)NH-; d is 0;

if E is E1 , o is an integer of 1-2;

if E is E2, o is an integer of 0-1 ;

if E is E3, o is 0;

G is Ci-3-alkyl; G5; G13;

R ²⁵ is H;

R ²⁶ , R ²⁷ , and R ²⁸ are independently H; F; CI; CH ₃ ; CF ₃ ; OCH ₃ ; OCF ₃ ; or OCHF ₂ ; R ²⁹ is H; F; CH ₃ ; or CF ₃ ;

R ³ ° is H; OH; or NH ₂ ; Tis N; orCH; with the proviso that

G13 is a group of one of the formulae

G13 ¹ G13" and with the proviso that,

if Q is O; S; or

if R ³⁴ in Q1 is H; F; CF ₃ ; OH; Ci-4-alkyl; C _2- -alkenyl; or Ci-4-alkoxy;

then GisG5;G13';orG13";

Qis 0;S;Q1;orQ2;

Z ³ is -NR ⁴⁸ C(0)-; -NR ⁴⁸ C(0)NR ⁴⁸ -; or -NR ⁸ S(0) ₂ -;

R ³⁴ is H; F; CF ₃ ; OH; Ci _-4 -alkyl; C ₂ - ₄ -alkenyl; or Ci _-4 -alkoxy;

R ³⁷ is H; F; CI; CH ₃ ; OCH ₃ ; CF ₃ ; OCF ₃ ; or OCHF ₂ ;

R ³⁸ is H; F; CI; CF ₃ ; OCF ₃ ; OCHF ₂ ; Ci-*-alkyl; C _2-4 -alkenyl; Ci- ₄ -alkoxy;

Ci _-4 -thioalkoxy; or C _3-4 -cycloalkyl;

R ³⁹ is H; F; CI; Br; I; CF ₃ ; OH; OCF ₃ ; OCHF ₂ ; CN; Ci- ₆ -alkyl; Ci- ₆ -alkoxy;

Ci-6-thioalkoxy; C ₃ -6-cycloalkyl; or C _3- 6-heterocyclyl;

R ³ is H13;

R ⁴⁴ is H; F; CI; CF ₃ ; OCF ₃ ; OCHF ₂ ; Ci--alkyl; or

C _3-4 -cycloalkyl;

R ⁵ is H; F; CI; or CH ₃ ;

R ⁴⁷ and R ⁴⁸ are H; m and n are 0;

e is 1 ;

g is 0;or1;

Uis 0;orCHR ⁴⁷ ; with the proviso that,

if i is 1 , and p is 1 ;

then Q is Q1 ; or Q2;

and with the further proviso that,

if G is Ci- ₃ alkyl;

then Q is Q2; and with the proviso that,

if i is 2, and p is 1 ;

then Q is O; S; or Q1 ; R ³⁴ in Q1 is H; the 6-membered (t=1 ) ring system positioned between functional moieties L and X is H24;

R ⁴⁹ , R ⁵⁰ and R ⁵² are independently H; F; CI; CH ₃ ; CF ₃ ; OCH ₃ ; OCF ₃ ; or OCHF ₂ ; R ⁵¹ is H; F; CI; CF ₃ ; OCF ₃ ; OCHF ₂ ; NH ₂ ; Ci _-3 -alkyl; or Ci _-3 -alkoxy; with the proviso that in H24 two of the substituents are H; or tautomers or rotamers thereof; or salts; or pharmaceutically acceptable salts; solvates thereof.

A further embodiment (4) of the invention relates to compounds of formula (I) according to embodiment (3),

wherein

in substituent E

R ¹⁷ is -C(0)OR ^18a ;

R ²² is H; or -(CHR ²³ ) ₀ C(0)OR ^18a ; in substituent Q

R ³⁴ is H; F; CF ₃ ; OH; Ci _-4 -alkyl; or Ci-4-alkoxy;

R ³⁶ is is a group of one of the formulae

H1 ¹ H2 ¹

R ³⁷ and R ³⁸ are H; F; CI; CH ₃ ; OCH ₃ ; CF ₃ ; OCF ₃ ; or OCHF ₂ ; R ³⁹ is H; F; CI; CF ₃ ; OH; OCF ₃ ; OCHF ₂ ; Ci-4-alkyl; Ci-4-alkoxy;

or Ci-4-thioalkoxy;

M' is 0; or S; or tautomers or rotamers thereof; or salts; or pharmaceutically acceptable salts; solvates thereof.

A further embodiment (5) of the invention relates to compounds of formula (I) according to embodiment (3),

wherein

E is a group of one of the formulae

-naphthyl; 2-naphthyl; or a group of one of the formulae

with the proviso that,

if Q is O, orQ1;

then G is phenyl; 1-naphthyl; 2-naphthyl; G5'; G5"; G5 ^m ; G5 ^IV ; G5 ^V ; G5 ^VI ; G5 ^VM ;

G ₅ VIII. _G13 III. orG13 ^lv ;

Qis 0;Q1;orQ2;

Z ³ is -NHC(O)-; or -NHC(0)NH-;

R ³⁴ is H;OH;orOCH ₃ ; 3 ⁶ is a group of one of the formulae

with the proviso that,

if i is 1 , and p is 1 ;

then Q is Q1 ; or Q2;

and with the further proviso that,

if G is CH ₃ ;

then Q is Q2; with the proviso that,

if i is 2, and p is 1 ;

then Q is O; or Q1 ; R ³⁴ in Q1 is H; in H29 positioned between functional moieties L and X

R ⁴⁹ , R ⁵⁰ , R ⁵¹ , and R ⁵² are H; or

R ⁴⁹ is F, R ⁵⁰ is H, R ⁵¹ is H, and R ⁵² is F; or

R ⁴⁹ is F, R ⁵⁰ is F, R ⁵¹ is H, and R ⁵² is H; or

R ⁴⁹ is H, R ⁵⁰ is H, R ⁵¹ is H, and R ⁵² is CI; or

R ⁴⁹ is CI, R ⁵⁰ is H, R ⁵¹ is H, and R ⁵² is H; or

R ⁴⁹ is OCHs, R ⁵⁰ is H, R ⁵¹ is H, and R ⁵² is H; or

R ⁴⁹ is H, R ⁵⁰ is H, R ⁵¹ is H, and R ⁵² is OCH ₃ ; or

R ⁴⁹ is H, R ⁵⁰ is H, R ⁵¹ is NH ₂ , and R ⁵² is H; or tautomers or rotamers thereof; or salts; or pharmaceutically acceptable salts; solvates thereof. A further embodiment (6) of the invention relates to compounds of formula (I) according to embodiment (5),

wherein

E is a group of one of the formulae E1 ¹ ; E1"; E1 ^m ; E3'; or E3";

G is phenyl; 1-naphthyl; 2-naphthyl; or a group of one of the formulae G5";

G5'"; G5 ^VI ; orG13 ^lv ;

Qis 0;Q1;orQ2;

Z ³ is -NHC(O)-;

R ³⁴ is H;orOCH ₃ ;

R ³⁶ is a group of one of the formulae H2 ^m ; H2 ^IX ; or H2 ^XI ; with the proviso that,

if i is 1, and p is 1;

then Q is Q1; orQ2; and with the proviso that,

if i is 2, and p is 1 ;

then Q is O; or Q1 ; R ³⁴ in Q1 is H; in H24 positioned between functional moieties L and X

R ⁴⁹ , R ⁵⁰ , R5 ⁱ , and R ⁵² are H; or tautomers or rotamers thereof; or salts; or pharmaceutically acceptable salts; or solvates thereof.

A further embodiment (7) of the invention relates to compounds of formula (I) according to embodiment (6),

wherein

Eis E1»; orE1 ^IM ;

R ^18a and R ^18b areF1; or tautomers or rotamers thereof; or salts; or pharmaceutically acceptable salts; or solvates thereof. A further embodiment (8) of the invention relates to compounds of formula (I) according to embodiment (6),

wherein

E is E1 ¹ ; E3 ¹ ; or E3";

R ^18a is CH ₃ ; or CH ₂ CH ₃ ; or tautomers or rotamers thereof; or salts; or pharmaceutically acceptable salts; or solvates thereof. A further embodiment (9) of the invention relates to compounds of formula (I) according to embodiment (6),

wherein

E is E3';

R ^18a is CH ₂ CH ₃ ; or tautomers or rotamers thereof; or salts; or pharmaceutically acceptable salts; or solvates thereof.

A further embodiment (10) of the present invention may also include compounds, which are identical to the compounds of formula (I), except that one or more atoms are replaced by an atom having an atomic mass number or mass different from the atomic mass number or mass usually found in nature, e.g. compounds enriched in ² H (D), ³ H, ¹¹ C, ¹⁴ C, ¹²⁷ l etc. These isotopic analogs and their pharmaceutical salts and formulations are considered useful agents in the therapy and/or diagnostic, for example, but not limited to, where a fine-tuning of in vivo half-life time could lead to an optimized dosage regimen.

A further embodiment (1 1) of the invention relates to compounds of formula (I) according to embodiment (1 ) which are selected from the group consisting of Ex.1 to Ex.8, the lUPAC names of which are shown in Table 01 b; or tautomers or rotamers thereof; or salts; or pharmaceutically acceptable salts; or solvates thereof. General approach for the preparation of macrocyclic compounds of the invention Constituent building blocks

The macrocyclic compounds of the invention, as described above, can formally be dissected into building blocks A, B, and C. Additionally, building block C can be divided into two appropriately substituted subunits c1 and c2.

Scheme 1 : Building blocks

Buildin block A Building block B

Building block A is based on appropriately substituted and protected divalent phenol or thiophenol derivatives. Building block B is corresponding to appropriately substituted and protected secondary aminoalcohols. For building block C, appropriately substituted subunits c1 and c2 are derived from suitably substituted and protected precursors, like, but not limited to, appropriately substituted and protected amino acids derivatives.

Synthesis of the building blocks

For access to building block A and subunit c2 a plethora of literature precedents exists, and corresponding synthetic approaches have also been described in WO201 1/014973 A2, WO201 1/015241 A1 and WO2013/139697 A1 , disclosing macrocyclic compounds. These three patent applications likewise include synthetic strategies that are applicable for the preparation of building blocks B and c1 , both comprising a cyclic secondary amine group. Additional synthetic pathways to c1 , well known in the literature, are described in this section.

Functional groups not involved in the formation of the macrocyclic backbone can be diversified by standard methods of organic synthesis, preferably by parallel/ combinatorial chemistry. These derivatization methods are well-known to those skilled in the art (selected references: A. R. Katritzky et al. (eds), Comprehensive Functional Group Transformations, Pergamon, 1995; S. Patai, Z. Rappoport (eds), Chemistry of Functional Groups, Wiley, 1999; J. March, Advanced Organic Chemistry, 4 ed., Wiley, 1992; D. Obrecht, J.M. Villalgordo (eds), Solid-Supported Combinatorial and Parallel Synthesis of Small-Molecular-Weight Compound Libraries, Pergamon, 1998; W. Bannwarth et al. (eds), Combinatorial Chemistry: From Theory to Application, 2 ed., Wiley-VCH 2006).

Additional synthetic strategies towards subunit c1

Pyrrolidine derivatives

Many pyrrolidine derivatives comprising a carboxy group in alpha-position to the cyclic amine can be accessed through commercially available building blocks or from building blocks that are accessible through well established synthesis strategies.

Commercial (25,3S)-3-hydroxyproline represents a very suitable building block that was also used for the preparation of macrocyclic compounds of this invention.

Furthermore, the corresponding carboxy and/or hydroxy group can be transformed into other functional moieties by procedures well known to a person skilled in the art.

Piperidine derivatives

Piperidine derivatives comprising a carboxy group in alpha position to the cyclic amine can be prepared by various routes. Efficient approaches for 3-hydroxypipecolic acid derivatives include a reaction sequence starting from 1 ,5-pentandiol (B. B. Ahuja, A. Sudalai, Tetrahedron Asymmetry 2015, 26(1), 24-28), use of readily available L-(+)-tartaric acid as starting material (S. P. Chavan eta!, Tetrahedron Lett. 2013 , 54(36), 4851 -4853), or reduction of appropriately substituted pyridine compounds (J. Drummond eta/., J. Med. Chem. 1989, 32(9), 21 16-2128).

Transformation of the corresponding carboxy and/or hydroxy groups in the above- mentioned derivatives into other functional moieties can be performed by established procedures that are well known to a person skilled in the art. Synthetic strategies to (acyloxy)alkyl ester moieties in substituent E

Synthetic routes to (acyloxy)alkyl ester moieties in moiety E of compounds of the invention, as described above, have been described in the literature and are well known to a person skilled in the art.

For preparation of (acyloxy)alkyl esters of phosphonic acids, selected references are given for access to macrocydic phosphonic acid esters 52aa, 52ab, 52ba, and 52bb, as described below.

Further procedures have been reported for synthetic access to (acyloxy)alkyl esters of carboxylic acids (selected references: D. Baldwin et a/., Eur. J. Med. Chem. 1982, 17(4), 297-300; S. B. Singh et a/., J. Med. Chem. 2014, 57(20), 8421-8444; E. Defossa et a/., Liebigs Annates 1996, / /, 1743-1749; WO2015/89137 A1 , Example 249).

General processes for the synthesis of macrocydic compounds of formula (I)

General procedures for the synthesis of libraries of macrocydic compounds of formula (I) are described below. It will be immediately apparent to those skilled in the art how these procedures have to be modified for the synthesis of individual macrocydic compounds.

The macrocydic compounds of the invention can be obtained by cyclization of suitable linear precursors which are derived from optionally substituted bifunctional phenols or thiophenols A, substituted amino alcohols B, and two building blocks forming C. If needed, further transformations can be performed.

Variable substituents can be introduced by pre- or postcyclative derivatization of one or more orthogonally protected attachment points (e.g. amino groups, carboxyl groups, hydroxyl groups) on B, C or A. Variable R-groups may also be introduced as side chain motifs of the subunits of building block C.

The macrocydic products of the invention can be prepared either in solution or on solid support.

The essential ring closure reaction may be performed between any of the building blocks; for example, macrocycles of formula (I) may be obtained by

Macrolactamization between C and B;

Macrolactamization between A and C;

Sulfonamide formation between A and C;

Urethane or urea formation between the two subunits of building block C; or - Arylether or arylthioether formation between A and B; Properties and usefulness

The macrocyclic compounds of the invention exhibit antiproliferative activity on various cancers cell lines. In addition, they can be metabolized to modulators of Pin1 and thus may be used in a wide range of applications in order to modulate Pin1 activity, leading to the desired therapeutic effect in man or in other mammals.

These compounds can be used as prodrugs, the term "prodrug" referring to a compound that can be converted in vivo into the parent active form. In many situations a prodrug may be favorable, and a prodrug-type approach, for example, can be utilized for acid-bearing compounds to mask the corresponding acid moiety, including carboxylic and phosphonic acids. For example, a prodrug would be a compound that is administered as an ester (representing the "prodrug") to improve cell-permeability. Inside the cell, the ester would then be metabolically hydrolyzed to the respective acid, representing the active entity. Conventional procedures for the selection and preparation of prodrug derivatives are described, for example, in H. Bundgaard, Design of Prodrugs, Elsevier, 1985; and in B. Testa and J. M. Mayer, The Hydrolysis of Carboxylic Acid Ester Prodrugs, in Hydrolysis in Drug and Prodrug Metabolism: Chemistry, Biochemistry, and Enyzmology, Chapter 8, Verlag Helvetica Chimica Acta, 2003.

Especially, the compounds of the invention can be used as agents for treating and/or preventing and/or delaying the onset of diseases, disorders or conditions related to abnormal cell growth, such as various cancers, such as breast cancer, prostate cancer, cervical cancer, lung cancer, liver cancer, esophageal cancer, and gastric cancer; or sarcoma, such as Ewing's sarcoma; or lymphoma, such as non-Hodgkin lymphoma; or leukemia, such as promyelotic leukemia or acute myeloid leukemia; inflammatory diseases, such as asthma, allergic pulmonary eosinophilia, acute respiratory distress syndrome, rheumatoid arthritis or inflammatory bowel diseases; or acute neurological disorders, such as stroke; or neurodegenerative diseases, such as Huntington's disease and frontotemporal dementia; or viral infections, such as HIV/AIDS; or infection with intracellular and extracellular pathogens, such as infections by Chlamydia trachomatis; or osteolytic bone diseases, such as periodontitis; nonalcoholic steatohepatitis; or nonalcoholic steatohepatitis; or cardivascular diseases, such as diabetic vascular disease or diabetic restenosis; or cardiac hypertrophy; or immune diseases or disorders, such as diabetes, multiple sclerosis, lupus, macrophage mediated tissue damage, gastritis, or myeloproliferative syndromes; or transplant rejection; or parasitic diseases, such as Theileriosis, such as lymphoproliferative Theileriosis caused by Theileria annulata or Theileria parva. They can be administered singly, as mixtures of several macrocyclic compounds of the invention, in combination with other anti cancer agents, or antiviral (e.g. anti-HIV) agents, or in combination with other pharmaceutically active agents. The macrocyclic compounds can be administered per se or as pharmaceutical compositions. The macrocyclic compounds of the invention may be administered per se or may be applied as an appropriate formulation together with carriers, diluents or excipients well known in the art.

Pharmaceutical compositions comprising macrocyclic compounds of the invention may be manufactured by means of conventional mixing, dissolving, granulating, coated tablet making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes. Pharmaceutical compositions may be formulated in conventional manner using one or more physiologically acceptable carriers, diluents, excipients or auxiliaries which facilitate processing of the macrocyclic compounds into preparations which can be used pharmaceutically. Proper formulation depends upon the method of administration chosen.

For topical administration the macrocyclic compounds of the invention may be formulated as solutions, gels, ointments, creams, suspensions, etc. as are well known in the art.

Systemic formulations include those designed for administration by injection, e.g. subcutaneous, intravenous, intramuscular, intrathecal or intraperitoneal injection, as well as those designed for transdermal, transmucosal, oral or pulmonary administration.

For injections, the macrocyclic compounds of the invention may be formulated in adequate solutions, preferably in physiologically compatible buffers such as Hink's solution, Ringer's solution, or physiological saline buffer. The solutions may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the macrocyclic compounds of the invention may be in powder form for combination with a suitable vehicle, e.g., sterile pyrogen free water, before use. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation as known in the art.

For oral administration, the compounds can be readily formulated by combining the active macrocyclic compounds of the invention with pharmaceutically acceptable carriers well known in the art. Such carriers enable the ester compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions etc., for oral ingestion by a patient to be treated. For oral formulations such as, for example, powders, capsules and tablets, suitable excipients include fillers such as sugars, such as lactose, sucrose, mannitol and sorbitol; cellulose preparations such as maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP); granulating agents; and binding agents. If desired, desintegrating agents may be added, such as cross linked polyvinylpyrrolidones, agar, or alginic acid or a salt thereof, such as sodium alginate. If desired, solid dosage forms may be sugar coated or enteric coated using standard techniques.

For oral liquid preparations such as, for example, suspensions, elixirs and solutions, suitable carriers, excipients or diluents include water, glycols, oils, alcohols, etc. In addition, flavoring agents, preservatives, coloring agents and the like may be added.

For buccal administration, the composition may take the form of tablets, lozenges, etc. formulated as usual.

For administration by inhalation, the macrocyclic compounds of the invention are conveniently delivered in form of an aerosol spray from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g. dichlorodifluoromethane, trichlorofluromethane, carbon dioxide or another suitable gas. In the case of a pressurized aerosol the dose unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of e.g. gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the macrocyclic compounds of the invention and a suitable powder base such as lactose or starch.

The compounds may also be formulated in rectal or vaginal compositions such as suppositories together with appropriate suppository bases such as cocoa butter or other glycerides. In addition to the formulations described above, the macrocyclic compounds of the invention may also be formulated as depot preparations. Such long acting formulations may be administered by implantation (e.g. subcutaneously or intramuscularly) or by intramuscular injection. For the manufacture of such depot preparations the macrocyclic compounds of the invention may be formulated with suitable polymeric or hydrophobic materials (e.g. as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble salts.

Other pharmaceutical delivery systems may be employed such as liposomes and emulsions well known in the art. Certain organic solvents such as dimethylsulfoxide may also be employed. Additionally, the macrocyclic compounds of the invention may be delivered using a sustained release system, such as semipermeable matrices of solid polymers containing the therapeutic agent (e.g. for coated stents). Various sustained release materials have been established and are well known by those skilled in the art. Sustained release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the therapeutic agent, additional strategies for protein stabilization may be employed. As the macrocyclic compounds of the invention may contain charged residues, they may be included in any of the above described formulations as such or as pharmaceutically acceptable salts. Pharmaceutically acceptable salts tend to be more soluble in aqueous and other protic solvents than are the corresponding free forms. In addition, the compounds of the present invention and their pharmaceutical acceptable salts may be used per se or in any appropriate formulation in morphological different solid state forms, which may or may not contain different amounts of solvent, e.g. hydrate.

The macrocyclic compounds of the invention, or compositions thereof, will generally be used in an amount effective to achieve the intended purpose. It is to be understood that the amount used will depend on a particular application.

For the use of treating or preventing diseases, disorders, or conditions with an etiology comprising, or associated with Pin1 , the macrocyclic compounds of the invention or compositions thereof, are administered or applied in a therapeutically effective amount. Determination of a therapeutically effective amount is well within the capacities of those skilled in the art, especially in view of the detailed disclosure provided herein.

For systemic administration, a therapeutically effective dose can be estimated initially from in vitro assays. For example, a dose can be formulated in animal models to achieve a circulating macrocyclic compounds concentration range that includes the GI50 (or IC50) as determined in an cell-based assay. Such information can be used to more accurately determine useful doses in humans. Initial dosages can also be determined from in vivo data, e.g. animal models, using techniques that are well known in the art. One having ordinary skill in the art could readily optimize administration to humans based on animal data.

Dosage amounts for applications as compounds that can be metabolized to agents for Pin1 modulation may be adjusted individually to provide plasma levels of the macrocyclic compounds of the invention which are sufficient to maintain the therapeutic effect. Therapeutically effective serum levels may be achieved by administering multiple doses each day.

In cases of local administration or selective uptake, the effective local concentration of the macrocyclic compounds of the invention may not be related to plasma concentration. One having the ordinary skill in the art will be able to optimize therapeutically effective local dosages without undue experimentation.

The amount of macrocyclic compounds of the invention administered will, of course, be dependent on the subject being treated, on the subject's weight, the severity of the affliction, the manner of administration and the judgement of the prescribing physician.

Normally, a therapeutically effective dose of the macrocyclic compounds of the invention described herein will provide therapeutic benefit without causing substantial toxicity.

Toxicity of the macrocyclic compounds of the invention can be determined by standard pharmaceutical procedures in experimental animals, e.g., by determining the LD50 (the dose lethal to 50% of the population) or the LD100 (the dose lethal to 100% of the population). The dose ratio between toxic and therapeutic effect is the therapeutic index. Compounds which exhibit high therapeutic indices are preferred. The data obtained from these animal studies can be used in formulating a dosage range that is not toxic for use in humans. The dosage of the macrocyclic compounds of the invention lies preferably within a range of circulating concentrations that include the effective dose with little or no toxicity. The dosage may vary within the range depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dose can be chosen by the individual physician in view of the patient's condition (see, e.g. E. Fingl etal. 1975, In: The Pharmacological Basis of Therapeutics, Ch.1 , p.1 ). The effective dosage of the compounds of the invention employed may vary depending on the particular compound or pharmaceutical preparation employed, the mode of administration and the severity and type of the condition treated. Thus, the dosage regimen is selected in accordance with factors including the route of administration and the clearance pathway, e.g. the renal and hepatic function of the patient. A physician, clinician or veterinarian skilled in the art can readily determine and prescribe the amount of the single compound of the invention required to prevent, ameliorate or arrest the progress of the condition or disease. Optimal precision in achieving concentrations of the compounds without toxicity requires a regimen based on the kinetics of the compounds' availability to the target sites. This involves a consideration of the distribution, equilibrium, and elimination of the compounds of the invention.

Examples

The following examples illustrate the invention in more detail but are not intended to limit its scope in any way. Before specific examples are described in detail the used abbreviations and applied general methods are listed.

Abbreviations

addn: addition, additional

ADDP: 1 ,1 '-(azodicarbonyl)dipiperidine

Alloc: allyloxycarbonyl

aq.: aqueous

Ar: aryl

Bn: benzyl

Boc: tert.-butoxycarbonyl

Boc ₂ 0: di-tert. -butyl pyrocarbonate; Boc anhydride, di-tert. -butyl dicarbonate br.: broad

Bu: butyl

t-Bu: tert.-butyl

Cbz: benzyloxycarbonyl

d: day(s) or doublet (spectral)

DTT: 1 ,4-dithio-DL-threitol

DIC: Λ,Λ '-diisopropylcarbodiimide

DMAP: 4-(dimethylamino)pyridine

DMBA: 1 ,3-dimethylbarbituric acid

DMF: dimethylformamide

DMSO: dimethyl sulfoxide

EDC: 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide

Et: ethyl

Et ₃ N: triethylamine

Et ₂ 0: diethyl ether

EtOAc: ethyl acetate

EtOH: ethanol

FC: flash chromatography

FI-MS: flow injection mass spectrometry

h: hour(s) HATU: C^(7-azabenzotriazol-1-yl)-A,A,A ',/V-tetramethyluronium hexa- fluorophosphate

Hepes: 2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid

HOAt: 1 -hydroxy-7-azabenzotriazole

i.v.: in vacuo

m: multiplet (spectral)

MeCN: acetonitrile

MeOH: methanol

Me: methyl

MOPS: 3-(4-morpholinyl)-1-propanesulfonic acid

Pd(PPh3) ₄ : tetrakis(triphenylphosphine)palladium(0)

PP i3: triphenylphosphine

prep.: preparative

i-Pr: isopropyl

i-Pr ₂ NEt: Nethyl-A/,/V-diisopropylamine

q: quartet (spectral)

quant.: quantitative

rt: room temperature

s: singlet (spectral)

sat.: saturated

soln: solution

TBAF : tetrabutylammonium fluoride

t: triplet (spectral)

TBDMS: tert.-butyldimethylsilyl

TBDMSCI: tert.-butyldimethylsilyl chloride

tert.: tertiary

TFA: trifluoroacetic acid

TFE: trifluoroethanol

THF: tetrahydrofuran

TLC: thinlayer chromatography

TMSCI: trimethylsilyl chloride, trimethylchlorosilane

Tris-HCI: tris(hydroxymethyl)aminomethane hydrochloride General Methods

TLC: Merck (silica gel 60 F254).

Flash chromatography (FC): Fluka silica gel 60 (0.04-0.063 mm) and Interchim Puriflash IR 60 silica gel (0.04-0.063 mm).

I. Analytical HPLC-MS methods:

Retention time (R _t ) in minutes (purity at 220 nm in %), m/z [M+H] ⁺

UV wavelength 220 nm, 254 nm

MS: Electrospray Ionization

Volume of injection: 5 μΙ_

Method 1

LC-MS: Agilent HP1 100 (DAD detector)

Column: Ascentis Express™ C18 2.7 pm, 3x50 mm (5381 1 U - Supelco Inc.) Mobile Phases: A: 0.1 % TFA in water; B: 0.085% TFA in MeCN

Column temperature: 55°C

Flow rate: 1 .3 mL/min

Gradient: 0-0.05 min: 97% A, 3% B; 2.95 min: 3% A, 97% B; 2.95-3.15 min: 3% A, 97% B; 3.17-3.23 min: 97% A, 3% B

Method 1a: m/z 95 - 1800, 2 sec; centroid mode, positive mode 40V

Method 1 b: m/z 95 - 1800, 2 sec; profile mode, positive mode 80V

Method 1c: m/z 95 - 1800, 2 sec; centroid mode, positive mode 20V

Method 1d: m/z 95 - 1800, 2 sec; profile mode, positive mode 40V

Method 1e: m/z 95 - 1800, 2 sec; profile mode, positive mode 20V

Method 2

LC-MS: Agilent HP1 100 (DAD detector)

Column: Ascentis Express™ C18 2.7 pm, 3x50 mm (5381 1 U - Supelco Inc.) Mobile Phases: A: NH4HCO3 1 mM in water - pH=10; B: MeCN

Column temperature: 55°C

Flow rate: 1 .3 mL/min

Gradient: 0-0.05 min: 97% A, 3% B; 2.95 min: 3% A, 97% B; 2.95-3.15 min: 3% A, 97% B; 3.17-3.23 min: 97% A, 3% B

Method 2a: m/z 95 - 1800, 2 sec; profile mode, positive mode 40V Method 2b: m/z 95 - 1800, 2 sec; centroid mode, negative mode 40V

Method 2c: m/z 95 - 1800, 2 sec; centroid mode, positive mode 40V

Method 3

LC-MS: Dionex Ultimate 3000 RS (DAD detector)

Column: Ascentis Express™ C18 2.7 pm, 2.1x50 mm (53822-U - Supelco Inc.) Mobile Phases: A: 0.1 % TFA in water; B: 0.085% TFA in MeCN

Column temperature: 55°C

Flow rate: 1 .4 mL/min

Gradient: 0-0.05 min: 97% A, 3% B; 2.80 min: 3% A, 97% B; 2.80-3.18 min: 3% A, 97% B; 3.20-3.30 min: 97% A, 3% B

Method 3a: m/z 95 - 1800, 2 sec; centroid mode, positive mode 40V

Method 3b: m/z 95 - 1800, 2 sec; profile mode, positive mode 40V Method 4

LC-MS: Agilent HP1 100 (DAD detector)

Column: Ascentis Express™ F5 2.7 pm, 3x50 mm (53576-U - Supelco Inc.) Mobile Phases: A: 0.1 %TFA in water; B: 0.085% TFA in MeCN

Column temperature: 55°C

Flow rate: 1 .3 mL/min

Gradient: 0-0.05 min: 70% A, 30% B; 2.95 min: 3% A, 97% B; 2.95-3.15 min: 3% A, 97% B; 3.17-3.23 min: 70% A, 30% B

Method 4a: m/z 95 - 1800, 2 sec; profile mode, positive mode 40V Method 5

LC-MS: Agilent HP1000

Column: Ascentis Express™ C18 2.7 pm, 3x50 mm (5381 1-U- Supelco Inc.) Mobile Phases: A: 0.1 % TFA in water; B: 0.085% TFA in MeCN

Column temperature: 55°C

Flow rate: 1 .3 mL/min

Gradient: 0-0.05 min: 97% A, 3% B; 3.4 min: 3% A, 97% B; 3.4-3.65 min: 3% A, 97% B; 3.67-3.7 min: 97% A, 3% B

Method 5a: m/z 95 - 1800, 1.5 sec; centroid mode, positive mode 40V

Method 5b: m/z 95 - 1800, 1.5 sec; profile mode, positive mode 40V Method 6

LC-MS: Ultimate RS 3000

Column: Ascentis Express™ C18 2.7 pm, 2.1x50 mm (53822-U - Supelco Inc.) Mobile Phases: A: 0.1 % TFA in water; B: 0.085% TFA in MeCN

Column temperature: 55°C

Flow rate: 1 .4 mL/min

Gradient: 0-0.05 min: 97% A, 3% B; 2.8 min: 3% A, 97% B; 2.8-3.18 min: 3% A, 97% B; 3.2-3.3 min: 97% A, 3% B

Method 6a: m/z 95 - 1800, 2 sec; profile mode, positive mode 40V

Method 6b: m/z 95 - 1800, 2 sec; centroid mode, positive mode 40V

Method 7

LC-MS: Agilent HP1 100 (DAD detector)

Column: Ascentis Express™ C8 2.7 pm, 3x100 mm (53852-U - Supelco Inc.) Mobile Phases: A: 0.1 % TFA in water; B: 0.085% TFA in MeCN

Column temperature: 55°C

Flow rate: 0.75 mL/min

Gradient: 0-0.1 min: 95% A, 5% B; 1 1.0 min: 15% A, 85% B; 1 1 .02 min: 3% A, 97% B 1 1.02-12.5 min: 3% A, 97% B; 12.52-13.45 min: 95% A, 5% B

Method 7a: m/z 95 - 2000, 2 sec; profile mode, positive mode 40V

Method 7b: m/z 95 - 2000, 2 sec; profile mode, positive mode 60V

Method 7c: m/z 95 - 2000, 2 sec; centroid mode, positive mode 40V

Flow rate: 1 .4 mL/min

Gradient: 0-0.1 min: 80% A, 20% B; 7.0 min: 3% A, 97% B; 7.0-7.5 min: 3% A, 97% B; 7.52-7.8 min: 80% A, 20% B

Method 7d: m/z 95 - 2000, 2 sec; centroid mode, positive mode 40V Method 8

LC-MS: Thermo scientific Ultimate 3000 RS

Column: Ascentis Express™ F5 2.7 pm, 2.1x50 mm (53567U - Supelco Inc.) Mobile Phases: A: 0.1 % TFA in water; B: 0.085% TFA in MeCN

Column temperature: 55°C

Flow rate: 1 .4 mL/min

Gradient: 0-0.05 min: 97% A, 3% B; 2.8 min: 3% A, 97% B; 2.8-3.18 min: 3% A, 97% B; 3.2-3.3 min: 97% A, 3% B Method 8a: m/z 95 - 1800, 2 sec; centroid mode, positive mode 40V Method 9

LC-MS: Thermo scientific Ultimate 3000 RS

Column: Ascentis Express™ C18 2.7 pm, 2.1x50 mm (53822-U - Supelco Inc.) Mobile Phases: A: NH4HCO3 1 mM in water - pH=10; B: MeCN

Column temperature: 55°C

Flow rate: 1 .4 mL/min

Gradient: 0-0.05 min: 97% A, 3% B; 2.8 min: 3% A, 97% B; 2.8-3.18 min: 3% A, 97% B; 3.2-3.3 min: 97% A, 3% B

Method 9a: m/z 95 - 1800 Da, 2 sec; centroid mode, positive mode 40V

II. Preparative HPLC methods: . Reverse Phase - Acidic conditions

Method 1a

Column: XBridge™ C18 5 pm, 30x150 mm (Waters AG)

Mobile phases:

A. 0.1 % TFA in water/MeCN 98/2 (v/v)

B. 0.1 % TFA in MeCN

2. Reverse Phase - Basic conditions

Method 2a

Column: XBridge™ C18 5 pm, 30x150 mm (Waters AG)

Mobile phases:

A. 10 mM NH4HCO3 in water pH 10/ MeCN 98/2 (v/v)

B MeCN

FI-MS: Agilent HP1 100 or Ultimate RS 3000; m/z [M+H] ⁺

NMR Spectroscopy: Bruker Avance 300, ¹ H-NMR (300 MHz) in the indicated solvent at ambient temperature. Chemical shifts δ in ppm, coupling constants J in Hz.

Specific Examples

In the examples below and if no other sources are cited, leading reference for standard conditions of protecting group manipulations (protection and deprotection) are 1 ) P.G.M. Wuts, T.W. Greene, Greene's Protective Groups in Organic Synthesis, John Wiley and Sons, 4th Edition, 2006; 2) P.J. Koncienski, Protecting Groups, 3rd ed., Georg Thieme Verlag 2005; and 3) M. Goodman (ed.), Methods of Organic Chemistry (Houben-Weyl), Vol E22a, Synthesis of Peptides and Peptidomimetics, Georg Thieme Verlag 2004.

Starting materials (Scheme 2):

Methyl 4-hydroxybenzoate (1 ) is commercially available.

(2S,4R)-Allyl 4-((tert.-butoxycarbonyl)amino)-2-(hydroxymethyl)pyrrolidine -1 - carboxylate (2) was prepared as described in the preceding patent application WO2013/139697 A1. (S)-Allyl 2-(hydroxymethyl)pyrrolidine-1 -carboxylate (3) was prepared as described in the preceding patent application WO2013/139697 A1 .

(2S,4R)-1 -Benzyl 2-methyl 4-hydroxypyrrolidine-1 ,2-dicarboxylate (4) is commercially available.

(2S,4R)-Allyl 4-((tert.-butyldimethylsilyl)oxy)-2-(hydroxymethyl)pyrrolidi ne-1- carboxylate (8):

At 0°C, tert.-butyldimethylsilyl chloride (4.45 g, 28.6 mmol) was added in portions to a soln of 4 (5.0 g, 17.9 mmol) and imidazole (3.65 g, 53.7 mmol) in DMF (50 mL). The mixture was allowed to warm to rt and stirred for 7 h. Aq. workup (Et.20, 1 M aq. NaH ₂ P0 ₄ soln, sat. aq. NaHCOs soln; Na ₂ S0 ₄ ) and FC (hexane / EtOAc) afforded 5 (7.4 g).

A soln of 5 (6.9 g, ca 16.8 mmol) in THF (40 mL) was added at 0°C to a suspension of LiBH ₄ (0.55 g, 25.2 mmol) in THF (100 mL). The mixture was allowed to stir at 0°C to rt for 16 h. The mixture was treated with 1 M aq. Nah PC soln and extracted with EtOAc. The organic phase was dried (Na2S0 ₄ ), filtered and concentrated. FC (hexane / EtOAc) afforded 6 (4.85 g, 80% over the two steps).

A soln of 6 (4.3 g, 1 1 .8 mmol) in MeOH (170 mL) was hydrogenated for 2 h at rt and normal pressure in the presence of palladium on activated charcoal (0.62 g; 10% Pd; moistened with ca 50% H ₂ 0). Filtration of the mixture through celite and evaporation of the volatiles yielded 7 (2.65 g, 97%). Allyl chloroformate (1 .85 mL, 16.9 mmol) was added at 0°C to a mixture of 7 (2.6 g, 1 1.2 mmol) in CH ₂ CI ₂ (100 mL) and sat. aq. NaHCOs soln (31.5 mL). The mixture was stirred at rt for 15 h followed by an aq. workup (CH2CI2, sat. aq. NaHCC>3 soln; Na ₂ S0 ₄ ) to give 8 (3.2 g, 91 %).

Data of 8: Ci ₅ H ₂₉ N0 ₄ Si (315.5). FI-MS: 316.3 ([M+H] ⁺ ). ¹ H-NMR (DMSO-d ₆ ): 5.85 (m, 1 H); 5.20 (qd, J = 1 .7, 17.3, 1 H); 5.1 1 (qd, J = 1.6, 10.5, 1 H); 4.67 (m, 1 H); 4.55 - 4.35 (m, 3 H); 3.78 (br. m, 1 H); 3.45 - 3.20 (m, 4 H); 1 .95 (br. m, 1 H); 1.78 (br. m, 1 H); 0.78 (s, 9 H); 0.00 (2 s, 6 H). (S)-Piperidin-2-ylmethanol (9) is commercially available.

(R)-Morpholin-3-ylmethanol hydrochloride (10) is commercially available.

(S)-Allyl 2-(hydroxymethyl)piperidine-1-carboxylate (11 ) was prepared by Alloc protection of the secondary amino group of 9 by allyl chloroformate in CH2CI2 in the presence of aq. NaHCC>3 soln applying standard conditions.

Data of 11 : C10H17NO3 (199.2). FI-MS: 200.6 ([M+H] ⁺ ). ¹ H-NMR (DMSO-d ₆ ): 5.92 (m, 1 H); 5.27 (td, J = 1.7, 17.2, 1 H); 5.17 (td, J = 1.5, 10.5, 1 H); 4.72 (t, J = 5.2, 1 H), 4.50 (d-like m, J = 5.1 , 2 H); 4.10 (m, 1 H); 3.86 (br. d, J ca 13.3, 1 H); 3.55 - 3.37 (m, 2 H); 2.81 (br. t, J ca 12.6, 1 H); 1 .76 (br. d, J ca 12.2, 1 H); 1.60 - 1 .24 (m, 5 H).

(R)-Allyl 3-(hydroxymethyl)morpholine-4-carboxylate (12) was prepared by Alloc protection of the secondary amino group of 10 by allyl chloroformate in CH2CI2 in the presence of aq. NaHCC>3 soln applying standard conditions.

Data of 12: C ₉ Hi ₅ N0 ₄ (201.2). FI-MS: 202.1 ([M+H] ⁺ ). ¹ H-NMR (DMSO-d ₆ ): 5.92 (m, 1 H); 5.29 (d, J = 17.2, 1 H); 5.18 (d, J = 10.5, 1 H); 4.88 (t, J = 5.3, 1 H); 4.54 (d-like m, J = 5.0, 2 H); 3.89 (br. d, J ca 1 1 .5, 1 H); 3.86 - 3.75 (m, 2 H); 3.67 - 3.58 (m, 2 H); 3.42 - 3.29 (m, 3 H); 3.08 (br. t, 1 H). (2S,3S)-3-Hydroxypyrrolidine-2-carboxylic acid (13) is commercially available.

(2S,3S)-Methyl 3-hydroxypyrrolidine-2-carboxylate hydrochloride (14 HCI):

At 0°C, SOCI2 (1 .0 mL) was slowly added to a suspension of 13 (1 .12 g, 8.5 mmol) in MeOH (20 mL). The resulting solution was stirred for 16 h and concentrated. The resulting solid was washed (Et ₂ 0) to give 14 HCI (1.54 g, 99%). Data of 14 HCI: C ₆ HnN0 ₃ HCI (145.1 + 36.5). ¹ H-NMR (DMSO-de): 9.58 (very br. s, NH ₂ ⁺ ); 5.98 (br. s, 1 H); 4.49 (br. s, 1 H); 4.16 (d, J = 2.5, 1 H); 3.77 (s, 3 H); ca 3.30 (2 H; superimposed by H ₂ 0 signal); 2.03 - 1.83 (m, 2 H). (2S,3S)-1-(Tert.-butoxycarbonyl)-3-hydroxypyrrolidine-2-carb oxylic acid (15) was prepared by Boc protection of the secondary amino group of 13 by di-tert. -butyl dicarbonate in dioxane in the presence of aq. Na2CC>3 soln applying standard conditions.

Data of 15: Ci ₀ Hi ₇ NO ₅ (231 .2). FI-MS: 230.1 ([M-H]-). ¹ H-NMR (DMSO-d ₆ ): 5.43 (br. s, 1 H); 4.23 (br. m, 1 H); 3.92 (d, J = 12.4, 1 H); 3.46 - 3.29 (m, 2 H); 1 .85 (m, 1 H); 1 .72 (m, 1 H); 1 .40, 1.34 (2 s, 9 H).

1 -Tert.-butyl 3,3-diethyl azetidine-1 ,3,3-tricarboxylate (16) is commercially available. Diethyl 1 -[(2S,3S)-3-hydroxypyrrolidine-2-carbonyl]azetidine-3,3-dica rboxylate (19): At rt, a soln of 16 (4.6 g, 15.2 mmol) in CH ₂ CI ₂ (80 mL) was treated with TFA (20 mL) for 90 min. The volatiles were evaporated. The residue was dissolved in toluene and concentrated to afford I7 CF3CO2H (5.9 g; contains residual TFA, purity ca 80%). Data of 17 CF ₃ C0 ₂ H: C9H15NO4 CF3CO2H (201 .2+1 14.0). FI-MS: 202.0 ([M+H] ⁺ ). ¹ H- NMR (DMSO-de): 9.13 (br. s, NH ₂ ⁺ ); 4.33 (br. s, 4 H); 4.23 (q, J = 7.1 , 4 H); 1.22 (t, J = 7.1 , 6 H).

EDC HCI (1 .38 g, 7.2 mmol) was added at 0°C to a solution of 15 (1.28 g, 5.5 mmol) and 17 TFA (2.4 g, ca 6.1 mmol) in pyridine (20 mL). The mixture was stirred at rt for 15 h. Aq. workup (EtOAc, 1 M aq. HCI soln, half-sat. aq. NaHC0 ₃ soln, H ₂ 0, sat. aq. NaCI soln; Na ₂ S0 ₄ ) and purification by FC (hexane / EtOAc) afforded 18 (1 .45 g, 63%).

At 0°C, a soln of 18 (1.45 g, 3.5 mmol) in CH ₂ CI ₂ (32 mL) was treated with TFA (8 mL). Stirring was continued at rt for 90 min. The volatiles were evaporated. The residue was dissolved in toluene and concentrated to afford 19 CF ₃ C0 ₂ H (1.69 g, quant, yield; contains residual TFA).

Data of 19 CF ₃ C0 ₂ H: Ci ₄ H ₂₂ N ₂ 0 ₆ CF ₃ C0 ₂ H (314.3+1 14.0). LC-MS (method 1 a): R _t = 1 .1 1 (93%), 315.1 ([M+H] ⁺ ). Synthesis of the allyl 2-(arylamino)acetates 22a - 22h Allyl 2-(phenylamino)acetate 22a:

Allyl chloroacetate (3.55 mL, 30.5 mmol) was slowly added to a soln of aniline (21a; 5.4 mL, 59.2 mmol) and tetrabutylammonium iodide (10.9 g, 29.7 mmol) in DMF/THF 1 :1 (140 mL). The mixture was stirred at 60°C for 16 h followed by aq. workup (EtOAc, sat. aq. NaHCOs soln; Na ₂ S0 ₄ ). Purification by FC (hexane / EtOAc) afforded 22a (4.1 g, 70%).

Data of 22a: CnHi ₃ N0 ₂ (191.2). LC-MS (method 5a): R _t = 2.02 (94%), 192.2([M+H] ⁺ ).

Allyl 2-(naphthalen-1 -ylamino)acetate 22b

In analogy, 22b (7.6 g, 73%) was obtained from allyl chloroacetate (5.0 mL, 43.1 mmol), 1-naphthylamine (21 b; 15.0 g, 104.7 mmol) and tetrabutylammonium iodide (16 g, 43.3 mmol) applying the procedure described for the synthesis of 22a.

Data of 22b: Ci ₅ Hi ₅ N0 ₂ (241.3). LC-MS (method 3a): R _t = 1 .76 (99%), 242.1 ([M+H] ⁺ ).

Allyl 2-(naphthalen-2-ylamino)acetate 22c

In analogy, 22c (0.54 g, 73%) was obtained from allyl chloroacetate (0.355 mL, 3.1 mmol), 2-naphthylamine (21c; 0.87 g, 6.1 mmol) and tetrabutylammonium iodide (1.1 g, 3.1 mmol) applying the procedure described for the synthesis of 22a.

Data of 22c: C15H15NO2 (241.3). LC-MS (method 1 a): R _t = 2.26 (89%), 242.1 ([M+H] ⁺ ).

Tert.-butyl 4-((2-(allyloxy)-2-oxoethyl)amino)benzoate 22d

Allyl chloroacetate (0.43 mL, 3.7 mmol) was added to a suspension of tert.-butyl 4- aminobenzoate (21d; 1.4 g, 7.2 mmol) and tetrabutylammonium iodide (1.34 g, 3.6 mmol) in DMF (20 mL). The mixture was stirred at 80°C for 20 h. More allyl chloroacetate (0.21 mL, 1.8 mmol) was added and stirring was continued at 90°C for 20 h followed by aq. workup (EtOAc, half-sat. aq. NaHCOs soln; Na2S0 ₄ ). Purification by FC (hexane / EtOAc / MeOH) afforded 22d (0.76 g, 72%).

Data of 22d: Ci ₆ H ₂ iN0 ₄ (291 .3). LC-MS (method 1 a): R _t = 2.28 (98%), 292.1 ([M+H] ⁺ ).

Allyl 2-((4-chloro-3-(trifluoromethyl)phenyl)amino)acetate 22e

Allyl chloroacetate (1 .55 mL, 13.2 mmol) was slowly added to a soln of 4-chloro-3- (trifluoromethyl)aniline (21e; 5.18 g, 26.5 mmol) and tetrabutylammonium iodide (4.89 g, 13.2 mmol) in DMF/THF 1 :1 (64 ml_). The mixture was stirred at 60°C for 20 h. More allyl chloroacetate (0.77 ml_, 6.6 mmol) was added at rt and stirring at 60°C was continued for 27 h. Aq. workup (EtOAc, half-sat. aq. NaHCC>3 soln, H ₂ 0) and FC (hexane / EtOAc) yielded 22e (2.35 g, 40%).

Data of 22e: C12H11 CIF3NO2 (293.7). LC-MS (method 1 a): R _t = 2.41 (95%), 294.0 ([M+H] ⁺ ).

Allyl 2-((4-fluorophenyl)amino)acetate 22f

The aminoacetate 22f (0.97 g, 94%) was obtained from allyl chloroacetate (0.57 ml_, 4.9 mmol), 4-fluoroaniline (21f; 1 .1 g, 9.9 mmol) and tetrabutylammonium iodide (1.83 g, 4.9 mmol) applying the procedure described for the synthesis of 22a.

Data of 22f: C11 H12FNO2 (209.2). LC-MS (methodl a): R _t = 1 .88 (98%), 210.0 ([M+H] ⁺ ). Allyl 2-((5-(benzyloxy)naphthalen-2-yl)amino)acetate 22g

ADDP (3.2 g, 12.56 mmol) was added at 0°C to a soln of 6-aminonaphthalen-1-ol (20, 1 .0 g, 6.28 mmol), benzyl alcohol (1 .0 ml_, 9.66 mmol) and PPh ₃ (3.5 g, 12.56 mmol) in CHCI3 (30 ml_). The mixture was stirred at rt for 16 h and concentrated. FC (hexane / EtOAc) and precipitation with EtOAc / hexane 1 :3 afforded 21g (1.09 g, 70%).

Allyl chloroacetate (0.25 ml_, 2.1 mmol) was slowly added to a soln of 21g (1 .06 g, 4.3 mmol) and tetrabutylammonium iodide (0.8 g, 2.1 mmol) in DMF/THF 1 :1 (42ml_). The mixture was stirred at 60°C for 48 h. Aq. workup (EtOAc, sat. aq. NaHCOs soln, H ₂ 0; Na ₂ S0 ₄ ) and FC (hexane / EtOAc) yielded 22g (0.36 g, 48%).

Data of 22g: C22H21 NO3 (347.4). LC-MS (method 1 a): R _t = 2.61 (96%), 348.1 ([M+H] ⁺ ).

Allyl 2-((3-(trifluoromethyl)phenyl)amino)acetate 22h

Allyl chloroacetate (0.8 mL, 6.75 mmol) was slowly added to a soln of 3- (trifluoromethyl)aniline (21 h; 2.0 g, 12.4 mmol) and tetrabutylammonium iodide (2.5 g, 6.75 mmol) in DMF/THF 1 :1 (42 mL). The mixture was stirred at 60°C for 48 h. Aq. workup (EtOAc, sat. aq. NaHCOs soln, H ₂ 0; Na ₂ S0 ₄ ) and FC (hexane / EtOAc) yielded 22h (0.73 g, 42%).

Data of 22h: C12H12F3NO2 (259.2). LC-MS (method 1 a): R _t = 2.26 (91 %), 301 .1 ([M+H+MeCN] ⁺ ), 260.0 ([M+H] ⁺ ). Synthesis of macrocyclic esters 33a, 33b, 33c, 37, and 38 (Scheme 3):

Synthesis of the arylether 23

ADDP (3.78 g, 15 mmol) in CHCI ₃ (10 mL) was added drop by drop to a soln of methyl 4-hydroxybenzoate (1 ; 1 .82 g, 12 mmol), alcohol 2 (3.0 g, 10 mmol) and PPh ₃ (3.93 g, 15 mmol) in CHC (60 mL). The mixture was stirred at rt for 2 h followed by aq. workup (CH2CI2, sat. aq. NaHCC>3 soln; Na2S0 ₄ ). The resulting crude product was suspended in CH2CI2 / hexane 2:8 and filtered. The filtrate was concentrated and purified by FC (hexane / EtOAc) to afford 23 (4.28 g, 98%).

Data of 23: C22H30N2O7 (434.5). LC-MS (method 5a): R _t = 2.62 (98%), 435.2 ([M+H] ⁺ ).

Synthesis of the acid 24

At 0°C, LiOH H2O (1 .7 g, 41 mmol) in H ₂ 0 (30 mL) was added to a soln of 23 (6.0 g, 14 mmol) in THF (60 mL) and MeOH (30 mL). The mixture was stirred at rt for 16 h, partially concentrated, acidified with 1 M aq. HCI soln and extracted with EtOAc. The organic phase was dried (Na2S0 ₄ ), filtered and concentrated to give 24 (5.9 g, 100%).

Data of 24: C21 H28N2O7 (420.5). LC-MS (method 1 c): R _t = 1.98 (96%), 421.2 ([M+H] ⁺ ). Synthesis of the amide 25

A soln of HATU (0.77 g, 2.0 mmol) in DMF (3 mL) was added to a soln of acid 24 (0.783 g, 1 .86 mmol) and the amine-hydrochloride 14 HCI (0.353 g, 1 .94 mmol) in DMF (12 mL), immediately followed by i-Pr ₂ NEt (1.0 mL, 5.8 mmol). The soln was stirred at rt for 16 h followed by an aq. workup (EtOAc, 1 M aq. HCI soln, H2O; Na ₂ S0 ₄ ) and FC (hexane / EtOAc) to give 25 (0.78 g, 77%).

Data of 25: C27H37N3O9 (547.6). LC-MS (method 1 e): R _t = 1 .89 (90%), 548.0 ([M+H] ⁺ ).

Synthesis of the carbamate 27a

At 0°C, a soln of 22a (0.6 g, 3.14 mmol) and Et ₃ N (0.62 mL, 4.45 mmol) in dry CH ₂ CI ₂ (5 mL) was slowly added to a soln of phosgene (20% in toluene, 1 .65 mL, 3.3 mmol) in CH2CI2 (5 mL). Stirring was continued for 45 min. The volatiles were evaporated and Et.20 was added to the residue. The resulting suspension was filtered. The filtrate was concentrated to give crude 26a (0.78 g).

A soln of 26a (0.262 g) in CH2CI2 (5 mL) was slowly added at 0°C to a soln of 25 (0.56 g, 1.02 mmol) and Et ₃ N (0.43 ml, 3.1 mmol) in CH ₂ CI ₂ (5 mL). DMAP (0.055 g, 0.45 mmol) was added and stirring was continued at 0°C for 10 min then at rt for 16 h. Aq. workup (CH2CI2, 1 M aq. HCI soln; Na2S0 ₄ ) and purification by FC (hexane / EtOAc) yielded 27a (0.245 g, 31 %).

Data of 27a: C ₃ 9H ₄₈ N ₄ Oi ₂ (764.8). LC-MS (method 1 d): R _t = 2.46 (96%), 765.4 ([M+H] ⁺ ).

Synthesis of the carbamate 27b

At 0°C, a soln of 22b (0.439 g, 1 .82 mmol) and Et ₃ N (0.355 mL, 2.55 mmol) in dry CH2CI2 (4 mL) was slowly added to a soln of phosgene (20% in toluene, 0.96 mL, 1 .82 mmol) in CH2CI2 (4 mL). Stirring was continued for 30 min. The volatiles were evaporated. The resulting material was suspended in dry Et.20 and filtered. The filtrate was concentrated to give crude 26b (0.57 g).

A soln of 25 (0.4 g, 0.73 mmol) and Et ₃ N (0.2 ml, 1 .43 mmol) in CH2CI2 (4 mL) was slowly added at 0°C to a soln of 26b (0.25 g, 0.82 mmol) in CH2CI2 (4 mL) followed by DMAP (0.025 g, 0.2 mmol). Stirring was continued at 0°C for 10 min then at rt for 16 h. More Et ₃ N (0.1 ml, 0.72 mmol), a soln of 26b (0.2 g, 0.65 mmol) in CH ₂ CI ₂ (0.7 mL) and DMAP (0.01 g) were added and stirring was continued for 6 h. Aq. workup (CH2CI2, 1 M aq. HCI soln; Na ₂ S0 ₄ ) and purification by FC (hexane / EtOAc) yielded 27b (0.318 g, 53%).

Data of 27b: C ₄₃ H ₅ oN ₄ Oi ₂ (814.9). LC-MS (method 1 d): Rt = 2.59 (98%), 815.2 ([M+H] ⁺ ).

Synthesis of the carbamate 27c

At 0°C, a soln of 22c (0.1 12 g, 0.46 mmol) and Et ₃ N (0.091 mL, 0.65 mmol) in dry CH2CI2 (2 mL) was slowly added to a soln of phosgene (20% in toluene, 0.25 mL, 0.50 mmol) in CH2CI2 (2 mL). Stirring was continued for 30 min. The volatiles were evaporated. The residue was dissolved in CH2CI2 and evaporated to give crude 26c. A soln of 25 (0.25 g, 0.45 mmol) and Et ₃ N (0.13 ml, 0.91 mmol) in CH2CI2 (2 mL) was slowly added at 0°C to a soln of 26c (0.141 g) in CH2CI2 (2 mL) followed by DMAP (17 mg, 0.014 mmol). Stirring was continued at 0°C for 10 min then at rt for 16 h. The volatiles were evaporated. Aq. workup (EtOAc, 1 M aq. HCI soln; Na2S0 ₄ ) and purification by FC (hexane / EtOAc) yielded 27c (0.16 g, 43%).

Data of 27c: C ₄₃ H ₅ oN ₄ Oi ₂ (814.9). LC-MS (method 5a): R _t = 2.82 (98%), 815.2 ([M+H] ⁺ ). Synthesis of the amino acid 28a

A soln of 27a (417 mg, 0.55 mmol) and DMBA (255 mg, 1.63 mmol) in degassed CH ₂ CI ₂ / EtOAc 1 :1 (1 1 mL) was treated with Pd(PPh ₃ ) ₄ (50 mg) for 2.5 h at rt. The resulting suspension was filtered. The solid was washed (Et.20) and dried i.v. to give 28a (353 mg, 100%).

Data of 28a: C ₃₂ H ₄₀ N ₄ Oio (640.7). LC-MS (method 1 d): Rt = 1.63 (84%), 641.4 ([M+H] ⁺ ).

Synthesis of the amino acid 28b

A soln of 27b (315 mg, 0.39 mmol) and DMBA (181 mg, 1.16 mmol) in degassed CH2CI2 / EtOAc 1 :1 (7.6 mL) was treated with Pd(PPh ₃ ) ₄ (36 mg) for 2.5 h at rt. The resulting suspension was filtered. The solid was washed (E.2O) and dried i.v. to give 28b (275 mg, 100%).

Data of 28b: C ₃ 6H ₄₂ N ₄ Oio (690.7). LC-MS (method 1 d): Rt = 1.72 (89%), 691.3 ([M+H] ⁺ ).

Synthesis of the amino acid 28c

A soln of 27c (158 mg, 0.19 mmol) and DMBA (91 mg, 0.58 mmol) in degassed CH2CI2 / EtOAc 1 :1 (3.6 mL) was treated with Pd(PPh ₃ ) ₄ (18 mg) for 2.5 h at rt. Et ₂ 0 was added to the resulting suspension. Centrifugation, removal of the supernatant and washing of the pellet (Et ₂ 0) gave 28c (127 mg, 94%).

Data of 28c: C ₃₆ H ₄₂ N ₄ Oio (690.7). LC-MS (method 5a): R _t = 1.87 (94%), 691.2 ([M+H] ⁺ ). Synthesis of the macrocyclic lactam 29a

At 1 10°C, a mixture of 28a (175 mg, 0.27 mmol) and i-Pr ₂ NEt (0.21 mL, 1 .23 mmol) in dry DMF (70 mL) was added over 2 h to a mixture of 2-chloro-1 -methylpyridinium iodide (221 mg, 0.87 mmol) in toluene (500 mL). Stirring was continued for 1 .5 h followed by partial evaporation of the volatiles and aq. workup (EtOAc, 1 M aq. HCI soln, H2O; Na2S0 ₄ ). Purification of the crude product by FC (hexane / EtOAc) afforded 29a (79 mg, 46%).

Data of 29a: C ₃₂ H ₃₈ N ₄ 09 (622.7). LC-MS (method 1 d): Rt = 1.99 (84%), 623.4 ([M+H] ⁺ ). Synthesis of the macrocyclic lactam 29b

In analogy, 29b (26 mg, 20%) was obtained from 28b (132 mg, 0.19 mmol) applying the procedure described for the synthesis of 29a.

Data of 29b: C36H40N4O9 (672.7). LC-MS (method 1 d): Rt = 2.15 (99%), 673.3 ([M+H] ⁺ ).

Synthesis of the macrocyclic lactam 29c

In analogy, 29c (43 mg, 35%) was obtained from 28c (125 mg, 0.18 mmol) applying the procedure described for the synthesis of 29a.

Data of 29c: C36H40N4O9 (672.7). LC-MS (method 1 d): Rt = 2.20 (99%), 673.3 ([M+H] ⁺ ).

Synthesis of the macrocyclic amine 30a

TFA (0.65 mL) was added at 0°C to a soln of 29a (1 10 mg, 0.16 mmol) in CH2CI2 (3 mL). Stirring was continued at 0°C to rt for 2.75 h followed by an aq. workup (CH2CI2, sat. aq. NaHCOs soln; Na ₂ S0 ₄ ) and purification by FC (CH ₂ CI ₂ / MeOH) to yield 30a (71 mg, 76%).

Data of 30a: C27H30N4O7 (522.5). LC-MS (method 2a): R _t = 1 .44 (90%), 523.3 ([M+H] ⁺ ).

Synthesis of the macrocyclic amine 30b

TFA (0.1 mL) was added at 0°C to a soln of 29b (45 mg, 0.067 mmol) in CH ₂ CI ₂ (2 mL). Stirring was continued at 0°C to rt for 2.5 h. More TFA (0.2 mL) was added at 0°C and stirring was continued at 0°C to rt for 2 h, followed by an aq. workup (CH2CI2, sat. aq. NaHCOs soln; Na ₂ S0 ₄ ) to yield 30b (40 mg, quant, yield).

Data of 30b: C31 H32N4O7 (572.6). LC-MS (method 2a): R _t = 1 .68 (94%), 573.2 ([M+H] ⁺ ).

Synthesis of the macrocyclic amide 31a

At 0°C, a soln of 3-chlorobenzoyl chloride (0.034 mL, 0.27 mmol) in CH2CI2 (0.5 mL) was added to a soln of 30a (70 mg, 0.13 mmol) and i-Pr ₂ NEt (0.046 mL, 0.27 mmol) in CH2CI2 (3 mL). Stirring was continued for 4 h at 0°C followed by an aq. workup (CH2CI2, sat. aq. NaHCOs soln; Na ₂ S0 ₄ ) and purification by FC (hexane / EtOAc) to give 31a (88 mg, 98%).

Data of 31a: C34H33CIN4O8 (661.1 ). LC-MS (method 1 d): R _t = 2.06 (98%), 661 .3 ([M+H] ⁺ ). Synthesis of the macrocyclic amide 31 b

In analogy, 31 b (42 mg, 93%; purification by prep. TLC (EtOAc)) was obtained from 30b (36 mg, 0.063 mmol) applying the procedure described for the synthesis of 31a. Data of 31 b: CssHssCIISUOs (71 1.2). LC-MS (method 1 d): Rt = 2.21 (99%), 71 1 .2 ([M+H] ⁺ ).

Synthesis of the macrocyclic amine 30c and amide 31c

A soln of HCI (4 M in dioxane; 1 mL) was added to a soln of 29c (58 mg, 0.086 mmol) in dioxane (2 mL). Stirring at rt was continued for 2.5 h. The volatiles were evaporated to afford crude 30c HCI (61 mg, used without further purification).

At 0°C, a soln of 3-chlorobenzoyl chloride (0.018 mL, 0.14 mmol) in CH ₂ CI ₂ (0.5 mL) was added to a soln of 30c HCI (53 mg, 0.087 mmol) and i-Pr ₂ NEt (0.06 mL, 0.35 mmol) in CH2CI2 (2 mL). Stirring was continued for 4 h at 0°C to rt followed by an aq. workup (CH2CI2, sat. aq. NaHCC>3 soln; Na2S0 ₄ ) and purification by prep. TLC (CH2CI2 / EtOAc) to give 31c (20 mg, 32%).

Data of 31c: CssHssCIISUOs (71 1 .2). LC-MS (method 1 d): R _t = 2.24 (92%), 71 1.2 ([M+H] ⁺ ).

Synthesis of the macrocyclic acid 32a

At 0°C, a soln of 31a (87 mg, 0.13 mmol) in THF / MeOH / H ₂ 0 2:1 :1 (4 mL) was treated with LiOH H ₂ 0 (8.3 mg, 0.20 mmol) for 1 .5 h followed by addn of 1 M aq. HCI soln to reach pH 3-4 (indicator paper). The mixture was extracted with EtOAc. The organic phase was dried (Na2S0 ₄ ), filtered and concentrated to give 32a (83 mg, 97%).

Data of 32a: C ₃ 3H ₃ iCIN ₄ 0 ₈ (647.1 ). LC-MS (method 1 d): Rt = 1 .83 (96%), 647.2 ([M+H] ⁺ ).

Synthesis of the macrocyclic acid 32b

In analogy, 32b (38 mg, 95%) was obtained from 31 b (41 mg, 0.058 mmol) applying the procedure described for the synthesis of 32a.

Data of 32b: C ₃ 7H ₃ 3CIN ₄ 0 ₈ (697.1 ). LC-MS (method 2a): R _t = 1 .37 (98%), 697.2 ([M+H] ⁺ ).

Synthesis of the macrocyclic acid 32c

At 0°C, a soln of 31c (19 mg, 0.027 mmol) in THF / MeOH / H ₂ 0 2:1 :1 (1.2 mL) was treated with LiOH H ₂ 0 (3.5 mg, 0.08 mmol) for 1 .5 h followed by acidification with 1 M aq. HCI soln and extraction with EtOAc. The organic phase was dried (Na2SC>4), filtered and concentrated to give 32c (17 mg, 92%).

Data of 32c: C37H33CIN4O8 (697.1 ). LC-MS (method 1 d): R _t = 2.04 (95%), 697.2 ([M+H] ⁺ ).

Synthesis of the macrocyclic ester 33a

At 0°C, i-Pr ₂ NEt (0.021 mL, 0.13 mmol) was added to a soln of HATU (38 mg, 0.1 mmol) and HOAt (14 mg, 0.10 mmol), 32a (18 mg, 0.028 mmol) and β-alanine methyl ester hydrochloride (12 mg, 0.083 mmol) in DMF (2 mL). Stirring at 0°C was continued for 4.5 h followed by an aq. workup (EtOAc, 1 M aq. HCI soln, H2O; Na ₂ S0 ₄ ) and purification by prep. TLC (EtOAc) to afford 33a (12.5 mg, 62%).

Data of 33a: C37H38CIN5O9 (732.2). LC-MS (method 1 d): Rt = 1 .96 (98%), 732.4 ([M+H] ⁺ ). Synthesis of the macrocyclic ester 33b

At 0°C, i-Pr ₂ NEt (0.016 mL, 0.095 mmol) was added to a soln of HATU (25 mg, 0.66 mmol) and HOAt (10 mg, 0.073 mmol), 32b (15 mg, 0.022 mmol) and β-alanine methyl ester hydrochloride (9 mg, 0.064 mmol) in DMF (1 mL). Stirring at 0°C was continued for 2 h followed by an aq. workup (EtOAc, 1 M aq. HCI soln, H2O; Na2S0 ₄ ) and purification by prep. TLC (EtOAc) to afford 33b (12 mg, 71 %).

Data of 33b: C41 H40CIN5O9 (782.2). LC-MS (method 1 d): Rt = 2.1 1 (99%), 782.1 ([M+H] ⁺ ).

Synthesis of the macrocyclic ester 33c

At 0°C, i-Pr ₂ NEt (0.014 mL, 0.08 mmol) was added to a soln of HATU (19.4 mg, 0.05 mmol) and HOAt (7.2 mg, 0.05 mmol), 32c (13 mg, 0.019 mmol) and β-alanine methyl ester hydrochloride (7.1 mg, 0.051 mmol) in DMF (1 mL). Stirring at 0°C was continued for 2 h followed by an aq. workup (EtOAc, 1 M aq. HCI soln, H2O; Na2S0 ₄ ) and purification by prep. TLC (EtOAc) to afford 33c (12 mg, 82%).

Data of 33c: C ₄ i H ₄₀ CIN ₅ O9 (782.2). LC-MS (method 5b): R _t = 2.31 (99%), 782.1 ([M+H] ⁺ ).

Synthesis of the macrocyclic acid 34

At 0°C, a soln of 29a (43 mg, 0.69 mmol) in THF / MeOH / H ₂ 0 2:1 :1 (2 mL) was treated with LiOH H ₂ 0 (4.5 mg, 0.1 1 mmol) for 1.5 h. The mixture was acidified by addn of 1 M aq. HCI soln and extracted with EtOAc. The organic phase was dried (Na ₂ S0 ₄ ), filtered and concentrated to give 34 (42 mg, 99%).

Data of 34: C31 H36N4O9 (608.6). LC-MS (method 1 d): R _t = 1 .73 (83%), 609.4 ([M+H] ⁺ ). Synthesis of the macrocyclic amide 35

At 0°C, i-Pr ₂ NEt (0.042 mL, 0.25 mmol) was added to a soln of HATU (67 mg, 0.18 mmol) and HOAt (26 mg, 0.19 mmol), 34 (41 mg, 0.067 mmol) and β-alanine methyl ester hydrochloride (24 mg, 0.17 mmol) in DMF (3 mL). Stirring at 0°C was continued for 4 h followed by an aq. workup (EtOAc, 1 M aq. HCI soln, H ₂ 0; Na ₂ S0 ₄ ) and purification by FC (hexane / EtOAc) to afford 35 (34 mg, 73%).

Data of 35: C35H43N5O10 (693.7). LC-MS (method 1 d): Rt = 1.88 (96%), 694.4 ([M+H] ⁺ ).

Synthesis of the macrocyclic amine 36

TFA (0.2 mL) was added at 0°C to a soln of 35 (33 mg, 0.048 mmol) in CH ₂ CI ₂ (2 mL). Stirring was continued at 0°C to rt for 3 h followed by an aq. workup (CH ₂ CI ₂ , sat. aq. NaHCOs soln; Na ₂ S0 ₄ ) to yield 36 (25 mg, 89%).

Data of 36: C30H35N5O8 (593.6). LC-MS (method 2a): R _t = 1 .39 (75%), 594.3 ([M+H] ⁺ ). Synthesis of the macrocyclic ester 37

At 0°C, 3-(trifluoromethyl)benzoyl chloride (0.0065 mL, 0.044 mmol) was added to a soln of 36 (13 mg, 0.022 mmol) and i-Pr ₂ NEt (0.0074 mL, 0.044 mmol) in CH ₂ CI ₂ (1 .5 mL). Stirring was continued for 4 h at 0°C followed by an aq. workup (CH ₂ CI ₂ , sat. aq. NaHCOs soln; Na ₂ S0 ₄ ) and purification by prep. TLC (EtOAc) to give 37 (14 mg, 82%).

Data of 37: CssHssFsNsOg (765.7). LC-MS (method 1 d): Rt = 2.05 (93%), 766.4 ([M+H] ⁺ ).

Synthesis of the macrocyclic ester 38

At 0°C, 3-fluorobenzoyl chloride (0.004 mL, 0.034 mmol) was added to a soln of 36

(10 mg, 0.017 mmol) and i-Pr ₂ NEt (0.0057 mL, 0.034 mmol) in CH ₂ CI ₂ (1.5 mL).

Stirring was continued for 4 h at 0°C followed by an aq. workup (CH ₂ CI ₂ , sat. aq.

NaHCOs soln; Na ₂ S0 ₄ ) and purification by prep. TLC (EtOAc) to give 38 (9 mg, 75%).

Data of 38: C37H38FN5O9 (715.7). LC-MS (method 1 d): Rt = 1.86 (97%), 716.4 ([M+H] ⁺ ). Synthesis of macrocyclic esters 49, 50, 51 , 52aa, 52ab, 52ba, and 52bb (Scheme 4):

Synthesis of the amine 39

A soln of HCI (4 M in dioxane, 1 1 1 mL) was added at 0°C to a soln of 23 (12.07 g, 27.8 mmol) in dioxane (57 mL). The mixture was allowed to stir at rt for 4 h. The volatiles were evaporated to give 39 HCI (10.0 g, 97%).

Data of 39 HCI: C17H22N2O5 HCI (334.4+36.5). LC-MS (method 1 a): R _t = 1.42 (97%), 334.9 ([M+H] ⁺ ). Synthesis of the amide 40

At 0°C, i-Pr ₂ NEt (19.4 mL, 1 14 mmol) was slowly added to a soln of amine 39 HCI (10.6 g, 28.6 mmol), 3-chlorobenzoic acid (5.37 g, 34.3 mmol), HATU (16.3 g, 42.9 mmol) and HOAt (5.84 g, 42.9 mmol) in DMF (100 mL). The soln was stirred at rt for 16 h followed by an aq. workup (EtOAc, H ₂ 0, 1 M aq. HCI soln, H ₂ 0, sat. aq. NaHCOs soln, sat. aq. NaCI soln; Na ₂ S0 ₄ ) and FC (hexane / EtOAc) to afford 40 (13.0 g, 96%).

Data of 40: C2 ₄ H ₂ 5CIN ₂ 06 (472.9). LC-MS (method 1 c): R _t = 2.31 (97%), 473.0 ([M+H] ⁺ ). Synthesis of the acid 41

At 0°C, a 1 M aq. LiOH soln (55 mL, 55 mmol) was slowly added to a soln of 40 (12.95 g, 27 mmol) in MeOH / THF 1 :1 (170 mL). The mixture was allowed to warm to rt and was then heated for 2 h to 60°C. The mixture was concentrated, acidified with 1 M aq. HCI soln and extracted with CHCI3. The organic phase - a milky suspension - was separated, treated with MeOH to get a clear soln, dried (Na2S0 ₄ ), filtered and concentrated to yield 41 (13 g, 100%).

Data of 41 : C23H23CIN2O6 (458.9). LC-MS (method 1 a): R _t = 2.00 (95%), 458.8 ([M+H] ⁺ ). Synthesis of the amide 42

A soln of HATU (1.32 g, 3.5 mmol) in DMF (2 mL) was added to a soln of acid 41 (1.52 g, 3.31 mmol) and the amine-hydrochloride 14 HCI (0.604 g, 3.33 mmol) in DMF (23 mL). The resulting soln was cooled to 0°C, followed by the slow addn of i-Pr2NEt (1.7 mL, 9.9 mmol). The soln was stirred at rt for 16 h followed by an aq. workup (EtOAc, 1 M aq. HCI soln, H ₂ 0, sat. aq. NaHCOs soln, sat. aq. NaCI soln; Na ₂ S0 ₄ ) and FC (EtOAc) to yield 42 (1 .83 g, 94%). Data of 42: C29H32CIN3O8 (586.0). LC-MS (method 1 d): Rt = 1 .91 (93%), 586.2 ([M+H] ⁺ ).

Synthesis of the carbamate 43d

At 0°C, a soln of 22d (0.296 g, 1 .02 mmol) and Et ₃ N (0.2 mL, 1 .43 mmol) in dry CH2CI2 (3 mL) was slowly added to a soln of phosgene (20% in toluene, 0.6 mL, 1 .14 mmol) in CH2CI2 (3 mL). Stirring was continued for 1 .5 h. More phosgene soln (20% in toluene, 0.1 mL, 0.19 mmol) and EtsN (0.05 mL, 0.36 mmol) were added and stirring at 0°C was continued for 30 min. The volatiles were evaporated. The residue was treated with Et.20 and filtered. The filtrate was concentrated to give crude 26d (0.455 g).

A soln of crude 26d (0.455 g) in CH2CI2 (4 mL) was cooled to 0°C. A soln of 42 (0.595 g, 1 .02 mmol) and Et ₃ N (0.18 ml, 1 .3 mmol) in CH2CI2 (4 mL) was slowly added, followed by DMAP (0.063 g, 0.516 mmol). The mixture was stirred at rt for 16 h. Silica gel was added to the mixture and the volatiles were evaporated. Purification by FC (hexane / EtOAc) afforded 43d (0.576 g, 62%).

Data of 43d: C46H51CIN4O13 (903.4). LC-MS (method 4a): R _t = 1 .88 (99%), 903.4 ([M+H] ⁺ ). Synthesis of the carbamate 43e

At 0°C, a soln of 22e (0.251 g, 0.85 mmol) and Et ₃ N (0.177 mL, 1.28 mmol) in dry CH2CI2 (8 mL) was slowly added to a soln of phosgene (20% in toluene, 0.54 mL, 1 .02 mmol) in CH ₂ CI ₂ (10 mL). Stirring was continued for 3 h. More Et ₃ N (0.177 mL, 1 .28 mmol) and phosgene soln (20% in toluene, 0.36 mL, 0.68 mmol) were added and the mixture was stirred for 1 h at 0°C. More phosgene soln (20% in toluene, 0.14 mL, 0.26 mmol) was added and stirring continued for 1 .5 h. The volatiles were removed. The residue was suspended in Et^O and filtered. The filtrate was concentrated to afford crude 26e (0.401 g).

A soln of crude 26e (0.401 g) in CH ₂ CI ₂ (10 mL) was cooled to 0°C. A soln of 42 (0.50 g, 0.85 mmol) and Et ₃ N (0.177 ml, 1.28 mmol) in CH2CI2 (8 mL) was slowly added, followed by DMAP (0.052 g, 0.427 mmol). The mixture was stirred at rt for 18 h. Silica gel was added to the mixture and the volatiles were evaporated. Purification by FC (hexane / EtOAc / EtOH) afforded 43e (0.534 g, 69%).

Data of 43e: C42H41 CI2F3N4O11 (905.7). LC-MS (method 1 a): R _t = 2.67 (98%), 905.3 ([M+H] ⁺ ). Synthesis of the macrocyclic lactam 45d

A soln of 43d (572 mg, 0.63 mmol) and DMBA (312 mg, 2.0 mmol) in degassed CH ₂ CI ₂ / EtOAc 1 :1 (14 mL) was treated with Pd(PPh ₃ ) ₄ (58 mg) for 2.5 h at rt. The volatiles were evaporated. The residue was suspended in Et^O and filtered, washed, (Et.20) and dried i.v. to give crude 44d (553 mg; used without further purification).

At 1 10°C a mixture of 44d (265 mg) and i-Pr ₂ NEt (0.23 mL, 1 .34 mmol) in dry DMF (35 mL) was added over 2 h to a mixture of 2-chloro-1 -methylpyridinium iodide (268 mg, 1 .05 mmol) in toluene (550 mL). Stirring was continued for 1.5 h followed by partial evaporation of the volatiles and aq. workup (EtOAc, 1 M aq. HCI soln, H ₂ 0). Purification of the crude product by FC (hexane / EtOAc) afforded 45d (90 mg, 39%). Data of 45d: C ₃₉ H ₄ iCIN ₄ Oio (761.2). LC-MS (method 1 d): Rt = 2.35 (97%), 761.3 ([M+H] ⁺ ).

Synthesis of the macrocyclic lactam 45e

A soln of 43e (524 mg, 0.58 mmol) and DMBA (316 mg, 2.0 mmol) in degassed CH2CI2 / EtOAc 1 :1 (14 mL) was treated with Pd(PPh ₃ ) ₄ (67 mg) for 5 h at rt. The volatiles were evaporated. The residue was suspended in Et^O and filtered, washed (Et.20) and dried i.v. to give crude 44e (559 mg; used without further purification). At 1 10°C a mixture of 44e (278 mg) and i-Pr ₂ NEt (0.26 mL, 1.5 mmol) in dry DMF (8 mL) was added over 2 h to a mixture of 2-chloro-1-methylpyridinium iodide (279 mg, 1 .09 mmol) in toluene (560 mL). Stirring was continued for 1.5 h followed by partial evaporation of the volatiles and aq. workup (EtOAc, 1 M aq. HCI soln, H ₂ 0). Purification of the crude product by FC (hexane / EtOAc) afforded 45e (1 15 mg, 52%).

Data of 45e: C35H31CI2F3N4O8 (763.5). LC-MS (method 1 a): R _t = 2.36 (95%), 763.2 ([M+H] ⁺ ).

Synthesis of the macrocyclic acid 46d

At 0°C, a soln of 45d (160 mg, 0.21 mmol) in THF / MeOH / H ₂ 0 2:1 :1 (6 mL) was treated with 1 M aq. soln of LiOH H ₂ 0 (0.21 mL, 0.21 mmol) for 8 h. More LiOH soln (0.032 mL, 0.032 mmol) was added after 8 h and again after 13 h and stirring at 0°C to rt was continued for further 6 h followed by addn of 1 M aq. HCI soln to reach pH 3- 4 (indicator paper). The mixture was extracted with EtOAc. The organic phase was dried (Na2S0 ₄ ), filtered and concentrated to give 46d (142 mg, containing ca 10% of starting material 45d; used without further purification; yield ca 80%). Data of 46d: C38H39CIN4O10 (747.2). LC-MS (method 1 d): Rt = 2.15 (90%), 747.2 ([M+H] ⁺ ).

Synthesis of the macrocyclic acid 46e

At 0°C, a soln of 45e (90 mg, 0.12 mmol) in THF / MeOH / H ₂ 0 2:1 :1 (4 mL) was treated with LiOH H ₂ 0 (7.5 mg, 0.18 mmol) for 8 h followed by addn of 1 M aq. HCI soln to reach pH 3-4 (indicator paper). The mixture was extracted with EtOAc. The organic phase was dried (Na2S0 ₄ ), filtered and concentrated to give 46e (81 mg, 92%).

Data of 46e: C ₃ 4H29Cl2F ₃ N ₄ 08 (749.5). LC-MS (method 1 d): Rt = 2.16 (92%), 749.3 ([M+H] ⁺ ).

Synthesis of the macrocyclic amide 47

At 0°C, i-Pr ₂ NEt (0.054 mL, 0.31 mmol) was added to a soln of HATU (96 mg, 0.25 mmol) and HOAt (33 mg, 0.24 mmol), 46d (58 mg, ca 0.07 mmol) and β-alanine methyl ester hydrochloride (31 mg, 0.22 mmol) in DMF (4 mL). Stirring at 0°C was continued for 4.5 h followed by an aq. workup (EtOAc, H2O) and purification by FC (hexane / EtOAc) to afford 47 (53 mg, 82%).

Data of 47: C ₄ 2H ₄ 6CIN ₅ 0ii (832.3). LC-MS (method 1 d): Rt = 2.26 (98%), 832.3 ([M+H] ⁺ ).

Synthesis of the macrocyclic ester 49

At 0°C, a soln of 47 (49 mg, 0.059 mmol) in CH ₂ CI ₂ (0.5 mL) was treated with TFA (0.25 mL) for 1 .5 h. The volatiles were evaporated. The residue was washed (Et.20) and dried to give 48 (55 mg; immediately used without further purification).

At 0°C, i-Pr ₂ NEt (0.05 mL, 0.29 mmol) was added to a soln of HATU (42 mg, 0.1 1 mmol) and HOAt (15 mg, 0.1 1 mmol), 48 (30 mg, 0.038 mmol) and ammonium chloride (9.7 mg, 0.18 mmol) in DMF (1 mL). Stirring at 0°C was continued for 2.5 h, followed by an aq. workup (EtOAc, H ₂ 0; Na2S0 ₄ ) and purification by FC (CH2CI2 / MeOH) to afford 49 (26 mg, 57%).

Data of 49: CssHsgCINeOio (775.2). LC-MS (method 1 d): Rt = 1.70 (99%), 775.3 ([M+H] ⁺ ).

Synthesis of the macrocyclic ester 50

At 0°C, i-Pr ₂ NEt (0.056 mL, 0.33 mmol) was added to a soln of HATU (88 mg, 0.23 mmol) and HOAt (32 mg, 0.23 mmol), 46e (50 mg, 0.066 mmol) and β-alanine methyl ester hydrochloride (28 mg, 0.20 mmol) in DMF (6 mL). Stirring at 0°C was continued for 5 h followed by an aq. workup (EtOAc, 1 M aq. HCI soln, aq. NaCI soln) and purification by FC (EtOAc) to afford 50 (46 mg, 83%).

Data of 50: CssHseC FsNsOg (834.6). LC-MS (method 1 a): R _t = 2.33 (98%), 834.4 ([M+H] ⁺ ).

Synthesis of the macrocyclic ester 51

Ester 51 (55 mg, 89%; purification by FC (EtOAc)) was obtained from 46e (55 mg, 0.073 mmol) and methyl azetidine-3-carboxylate hydrochloride (33 mg, 0.22 mmol) applying the conditions outlined for the synthesis of 50.

Data of 51 : C39H36CI2F3N5O9 (846.6). LC-MS (method 1 a): R _t = 2.23 (92%), 846.2 ([M+H] ⁺ ).

Synthesis of the macrocyclic phosphonic acid 52a

At rt, i-Pr ₂ NEt (0.086 mL, 0.50 mmol) and chlorotrimethylsilane (0.049 mL, 0.039 mmol) were added to a suspension of (2-aminoethyl)phosphonic acid (24 mg, 0.19 mmol) in dry CH2CI2 (3.5 mL). The mixture was stirred for 2.5 h and then cooled to 0°C. HOAt (10.5 mg, 0.077 mmol), DIC (0.03 mL, 0.19 mmol) and 46e (29 mg, 0.039 mmol) in CH2CI2 (4.5 mL) were added. The mixture was allowed to stir at rt for 20 h, treated with 1 M aq. HCI soln and sat. aq. NaCI soln and extracted repeatedly with EtOAc. The organic phase was dried (Na2S0 ₄ ), filtered and concentrated. Purification of the crude product by prep. HPLC (method 1 a) afforded 52a (14 mg, 42%).

Data of 52a: CseHssClzFsNsOioP (856.6). LC-MS (method 3a): R _t = 1.53 (92%), 856.2 ([M+H] ⁺ ).

Synthesis of the macrocyclic phosphonic acid 52b

52b (18.5 mg, 47%; purification by prep. HPLC (method 1 a)) was obtained from 46e (35 mg, 0.047 mmol) and (aminomethyl)phosphonic acid (26 mg, 0.23 mmol) following the procedure outlined for the synthesis of 52a.

Data of 52b: C35H33CI2F3N5O10P (842.5). LC-MS (method 2a): R _t = 1.26 (94%), 842.1 ([M+H] ⁺ ).

Synthetic access to macrocyclic phosphonic acid esters 52aa, 52ab, 52ba, and 52bb Procedures for the preparation of (acyloxy)alkyl esters of phosphonic acids have been described in the literature and are well known to a person skilled in the art (selected references: R. B. Baudy et a/., J. Med. Chem. 2009, 52(3), J. K. Dickson et a/., J. Med. Chem. 1996, 39, 661-664; S. De Lombaert et a/., J. Med. Chem. 1994, 37, 498-51 1 ; H. T. Serafinowska et al. J. Med. Chem. 1995, 38, 1372- 1379). In particular, approaches for preparation of bis(pivaloyloxymethyl) esters (WO2004/087720 A1 ; Example 100) and bis(pivaloyloxyethyl) esters (WO2009/069100 A1 ; Example 50) are available, and can be based on the use of commercially available pivaloyloxymethyl chloride and 1-pivaloyloxyethyl chloride, respectively.

Synthesis of final products Ex.1 - Ex.2 (Scheme 5):

Synthesis of the amide 53

At 0°C, i-Pr ₂ NEt (1.0 mL, 5.84 mmol) was added to a soln of HATU (0.852 g, 2.24 mmol), 41 (0.857 g, 1 .86 mmol) and 19 CF ₃ C0 ₂ H (0.80 g, 1.86 mmol) in DMF (16 mL). Stirring was continued at rt for 20 h followed by an aq. workup (EtOAc, 1 M aq. HCI soln, sat. aq. NaHCC>3 soln, H ₂ 0, sat. aq. NaCI soln; Na2S0 ₄ ) and purification by FC (hexane / EtOAc then EtOAc / MeOH) to afford 53 (1.18 g, 84%; used without further purification).

Data of 53: C ₃ 7H ₄₃ CIN ₄ 0n (755.2). LC-MS (method 1 a): R _t = 2.03 (74%), 755.3 «M+H] ⁺ ).

Synthesis of the carbamate 54e

At 0°C, a soln of 22e (0.086 g, 0.29 mmol) and Et ₃ N (0.06 mL, 0.42 mmol) in dry CH2CI2 (2 mL) was slowly added to a soln of phosgene (20% in toluene, 0.2 mL, 0.38 mmol) in CH2CI2 (2 mL). Stirring was continued for 3 h. The volatiles were removed. The residue was suspended in Et.20 and filtered. The filtrate was concentrated to afford crude 26e, which was dissolved in CH2CI2 (2 mL) and cooled to 0°C. A soln of 53 (0.22 g, 0.29 mmol) and Et ₃ N (0.07 ml, 0.51 mmol) in CH2CI2 (2 mL) was slowly added, followed by DMAP (0.018 g, 0.15 mmol). The mixture was stirred at rt for 18 h. Silica gel was added to the mixture and the volatiles were evaporated. Purification by FC (hexane / EtOAc then EtOAc / MeOH) afforded 54e (0.09 g, 29%).

Data of 54e: C ₅ oH52Cl2F ₃ N ₅ Oi ₄ (1074.9). LC-MS (method 7b): R _t = 10.12 (90%), 1074.3 ([M+H] ⁺ ).

Synthesis of the carbamate 54h

At 0°C, a soln of 22h (0.125 g, 0.48 mmol) and Et ₃ N (0.10 mL, 0.73 mmol) in dry CH2CI2 (5 mL) was slowly added to a soln of phosgene (20% in toluene, 0.38 mL, 0.72 mmol) in CH2CI2 (5 mL). Stirring was continued for 1 h. The volatiles were removed. The residue was suspended in Et ₂ 0 and filtered. The filtrate was concentrated to afford crude 26h, which was dissolved in CH2CI2 (5 mL) and cooled to 0°C. A soln of 53 (0.5 g, 0.48 mmol) and Et ₃ N (0.10 ml, 0.73 mmol) in CH2CI2 (5 mL) was slowly added, followed by DMAP (0.03 g, 0.25 mmol). The mixture was stirred at rt for 16 h. Silica gel was added to the mixture and the volatiles were evaporated. Purification by FC (hexane / EtOAc then EtOAc / MeOH) afforded 54h (0.212 g, 42%).

Data of 54h: C ₅ oH ₅ 3CIF ₃ N ₅ Oi4 (1040.4). LC-MS (method 6b): R _t = 2.23 (91 %), 1040.4 ([M+H] ⁺ ).

Synthesis of the amino acid 55e

A soln of 54e (85 mg, 0.079 mmol) and DMBA (44 mg, 0.28 mmol) in degassed CH2CI2 / EtOAc 1 :1 (2 mL) was treated with Pd(PPh ₃ ) ₄ (9 mg) for 1 h at rt. The volatiles were evaporated. The residue was suspended in Et ₂ 0 and filtered, washed (Et ₂ 0) and dried i.v. Purification by FC (CH ₂ CI ₂ / MeOH) gave 55e (60 mg, 80%). Data of 55e: C43H44CI2F3N5O12 (950.7). LC-MS (method 1 a): R _t = 2.01 (95%), 950.2 ([M+H] ⁺ ). Synthesis of the amino acid 55h

In analogy, 55h (142 mg, 77%; purification by FC (CH2CI2 / MeOH)) was obtained from 54h (210 mg, 0.20 mmol) applying the procedure outlined for the synthesis of 55e.

Data of 55h: C43H ₄₅ CIF ₃ N50i2 (916.3). LC-MS (method 9a): R _t = 1.33 (91 %), 916.3 ([M+H] ⁺ ).

Synthesis of the macrocyclic ester Ex.1

At 1 10°C, a soln of 55e (35 mg, 0.037 mmol) and i-Pr ₂ NEt (0.03 mL, 0.175 mmol) in dry DMF (4 mL) was added over 2 h to a mixture of 2-chloro-1 -methylpyridinium iodide (35 mg, 0.137 mmol) in toluene (76 mL). Stirring was continued for 1 h followed by partial evaporation of the volatiles and aq. workup (EtOAc, 1 M aq. HCI soln, H2O). Purification of the crude product by FC (hexane / EtOAc) afforded Ex.1 (8 mg, 23%).

Data of Ex.1 : cf. Table 01a

1 H-NMR (DMSO-de): 8.62 (d, J = 6.7, 1 H); 7.98 (s, 1 H); 7.88 (t, J = 1 .8, 1 H); 7.81 - 7.78 (m, 3 H); 7.60 (d-like m, 1 H); 7.50 (t, J = 7.85, 1 H); 7.25 (very br. s, 2 H); 7.05 (very br. s, 2 H); 5.31 (d, J = 1 1 .8, 1 H); 4.87 (d, J = 5.6, 1 H); 4.80 - 4.72 (m, 2 H); 4.68 (q, J = 6.9, 1 H); 4.53 (d, J = 1 1 .4, 1 H); 4.51 (s, 1 H); ca 4.30 - 4.02 (several m, 8 H); 3.96 (q-like m, 1 H); 3.81 (br. dd, J ca 6.9, 10.3, 1 H); 3.59 (dd, J = 7.1 , 10.6, 1 H); 3.44 (d, J = 14.9, 1 H); 3.36 (m, 1 H, partially superimposed by H ₂ 0 signal); ca 2.4 (m, 1 H); 2.26 - 2.00 (2 m, 2 H); 1 .69 (m, 1 H); 1.22 (t, J = 7.1 , 3 H); 1.10 (t, J = 7.1 , 3 H).

Synthesis of the macrocyclic ester Ex.2

At 90°C, a soln of 55h (70 mg, 0.076 mmol) and i-Pr ₂ NEt (0.065 mL, 0.38 mmol) in dry DMF (25 mL) was added over 2 h to a mixture of 2-chloro-1 -methylpyridinium iodide (69 mg, 0.27 mmol) in toluene (500 mL). Stirring was continued for 1 h followed by partial evaporation of the volatiles and aq. workup (EtOAc, 1 M aq. HCI soln, H2O). Purification of the crude product by FC (hexane / EtOAc / MeOH) afforded Ex.2 (10 mg, 14%).

Data of Ex.2: cf . Table 01a

Synthesis of the macrocyclic dicarboxylic acid 56e

At 0°C a soln of Ex.1 (10 mg, 0.01 1 mmol) in THF / EtOH 4:1 (1 .0 mL) was treated with a 2 M aq. soln of LiOH H ₂ 0 (0.025 mL, 0.05 mmol) for 4 h. The mixture was then stirred for 2 h at rt. More 2 M aq. soln of LiOH H ₂ 0 (0.025 mL, 0.05 mmol) was added and stirring was continued for 2 h. More 2 M aq. soln of LiOH H ₂ 0 (0.025 mL, 0.05 mmol) was added and stirring was continued for 0.5 h, followed by addn of 1 M aq. H3PO4 soln and extraction with EtOAc. The organic phase was dried (Na ₂ S0 ₄ ), filtered and concentrated. Purification of the crude product by prep. HPLC (method 2a) afforded 56e (5.0 mg, 51 %).

Data of 56e: Css^C FsNsOn (876.6). LC-MS (method 1 d): R _t = 1 .99 (90%), 876.1 ([M+H] ⁺ ). H-NMR (DMSO-de): 8.57 (d, J = 5.5, 1 H); 7.95 (br. s, 1 H); 7.91 (br. s, 1 H); 7.83 (d, J = 7.6, 1 H); 7.76 (s, 2 H); 7.58 (d-like m, 1 H); 7.48 (t, J = 7.9, 1 H); 7.24 (very br. s, 2 H); 7.07 (very br. s, 2 H); 5.29 (br. d, J = 12.2, 1 H); 4.88 - 4.68 (m, 3 H); 4.65 - 4.50 (m, 3 H); ca 4.20 - 3.70 (several m, 7 H); ca 3.70 - 3.30 (2 H, superimposed by H ₂ 0 signal); 2.35 (m, 1 H); 2.25 (m, 1 H); 2.12 (m, 1 H); 1.70 (m, 1 H).

Synthesis of the macrocyclic dicarboxylic acid 56h

A soln of Ex.2 (13 mg, 0.014 mmol) in THF / EtOH 4:1 (1 .25 mL) was treated with a 2 M aq. soln of LiOH H ₂ 0 (0.045 mL, 0.09 mmol) for 2.5 h at rt, followed by addn of 1 M aq. HCI soln (0.1 mL). The volatiles were evaporated. The solid residue was suspended in EtOAc and filtered. The filtrate was concentrated to afford 56h (1 1 mg, 90%).

Data of 56h: C39H35CIF3N5O11 (842.2). LC-MS (method 6a): R _t = 1.49 (91 %), 842.2 ([M+H] ⁺ ).

Synthesis of final products Ex.3 - Ex.6 (Scheme 6):

Synthesis of the acid 58

ADDP (4.09 g, 16.2 mmol) in CHCI ₃ (10 mL) was added drop by drop at 0°C to a soln of methyl 4-hydroxybenzoate (1 ; 1.97 g, 13 mmol), alcohol 3 (2.0 g, 10.8 mmol) and PP i3 (4.25 g, 16.2 mmol) in degassed CHCI3 (30 mL). The mixture was stirred at rt for 16 h followed by aq. workup (CH2CI2, sat. aq. NaHCC soln; Na2S0 ₄ ). The resulting crude product was suspended in CH2CI2 / hexane 2:8 and filtered. The filtrate was concentrated and purified by FC (hexane / EtOAc) to afford 57 (3.84 g, contaminated with ca 18% of 1 ; used without further purification).

At 0°C, 2 M aq. LiOH soln (14.7 mL, 29.4 mmol) was added to a soln of 57 (3.8 g, ca 9.8 mmol) in THF (60 mL) and MeOH (30 mL). The mixture was stirred at rt for 3 h. More 2 M aq. LiOH soln (7 mL, 14 mmol) was added and stirring was continued for 4 h, followed by partial evaporation of the volatiles, dilution with 1 M aq. HCI soln (150 mL) and extraction with CHCI3. MeOH was added to the organic phase to dissolve precipitating material. The organic phase was dried (Na2S0 ₄ ), filtered and concentrated. Purification of the crude product by FC (hexane / EtOAc) gave 58 (1 .93 g, 58%).

Data of 58: Ci ₆ Hi ₉ N0 ₅ (305.3). LC-MS (method 1 a): R _t = 1.82 (98%), 306.1 ([M+H] ⁺ ). Synthesis of the amide 59

At 0°C, i-Pr ₂ NEt (2.4 mL, 14 mmol) was added to a soln of HATU (2.1 g, 5.55 mmol), 58 (1.41 g, 4.62 mmol) and 19 CF ₃ C0 ₂ H (2.2 g, 4.62 mmol) in DMF (40 mL). Stirring was continued at rt for 3 h followed by an aq. workup (EtOAc, 1 M aq. HCI soln, sat. aq. NaHCOs soln, H2O, sat. aq. NaCI soln; Na2S0 ₄ ) and purification by FC (hexane / EtOAc then EtOAc / MeOH) to afford 59 (2.4 g, 86%).

Data of 59: C30H39N3O10 (601 .6). LC-MS (method 1 a): R _t = 1.88 (80%), 602.2 ([M+H] ⁺ ). Synthesis of the carbamate 60a

At 0°C, a soln of 22a (0.095 g, 0.5 mmol) and Et ₃ N (0.1 ml_, 0.72 mmol) in dry CH ₂ CI ₂ (3 ml.) was slowly added to a soln of phosgene (20% in toluene, 0.4 ml_, 0.76 mmol) in CH2CI2 (3 ml_). Stirring was continued for 1 h. The volatiles were removed. The residue was suspended in Et.20 and filtered. The filtrate was concentrated to afford crude 26a, which was dissolved in CH2CI2 (3 ml.) and cooled to 0°C. A soln of 59 (0.30 g, 0.5 mmol) and Et ₃ N (0.1 ml, 0.72 mmol) in CH2CI2 (3 ml.) was slowly added, followed by DMAP (0.03 g, 0.25 mmol). The mixture was stirred at rt for 18 h. Silica gel was added to the mixture and the volatiles were evaporated. Purification by FC (hexane / EtOAc then EtOAc / MeOH) afforded 60a (0.16 g, 39%).

Data of 60a: C42H50N4O13 (818.9). LC-MS (method 8a): R _t = 1 .86 (92%), 819.2 ([M+H] ⁺ ).

Synthesis of the carbamate 60e

At 0°C, a soln of 22e (0.18 g, 0.61 mmol) and Et ₃ N (0.1 ml_, 0.72 mmol) in dry CH ₂ CI ₂ (4 ml.) was slowly added to a soln of phosgene (20% in toluene, 0.46 ml_, 0.87 mmol) in CH2CI2 (4 ml_). Stirring was continued for 1 h. The volatiles were removed. The residue was suspended in Et^O and filtered. The filtrate was concentrated to afford crude 26e, which was dissolved in CH2CI2 (4 ml.) and cooled to 0°C. A soln of 59 (0.35 g, 0.58 mmol) and Et ₃ N (0.1 ml, 0.72 mmol) in CH2CI2 (4 ml.) was slowly added, followed by DMAP (0.035 g, 0.29 mmol). The mixture was stirred at rt for 18 h. Silica gel was added to the mixture and the volatiles were evaporated. Purification by FC (hexane / EtOAc then EtOAc / MeOH) afforded 60e (0.22 g, 41 %).

Data of 60e: C43H48CIF3N4O13 (921 .3). LC-MS (method 8a): R _t = 2.07 (92%), 921.3 ([M+H] ⁺ ).

Synthesis of the carbamate 60f

Carbamate 60f (0.18 g, 43%; purification by FC (hexane / EtOAc then EtOAc / MeOH) was obtained from 22f (0.104 g, 0.5 mmol) and 59 (0.30 g, 0.5 mmol) by following the procedure outlined for the synthesis of 60a.

Data of 60f: C42H49FN4O13 (836.9). LC-MS (method 8a): R _t = 1 .90 (92%), 837.3 ([M+H] ⁺ ).

Synthesis of the carbamate 60g

At 0°C, a soln of 22g (0.23 g, 0.67 mmol) and Et ₃ N (0.14 mL, 1.0 mmol) in dry CH ₂ CI ₂ (5 mL) was slowly added to a soln of phosgene (20% in toluene, 0.875 mL, 1 .66 mmol) in CH2CI2 (5 ml_). Stirring was continued for 1 h. The volatiles were removed. The residue was suspended in Et ₂ 0 and filtered. The filtrate was concentrated to afford crude 26g, which was dissolved in CH2CI2 (5 ml.) and cooled to 0°C. A soln of 59 (0.5 g, 0.83 mmol) and Et ₃ N (0.14 ml, 1.0 mmol) in CH2CI2 (5 ml.) was slowly added, followed by DMAP (0.04 g, 0.33 mmol). The mixture was stirred at rt for 18 h. Silica gel was added to the mixture and the volatiles were evaporated. Purification by FC (hexane / EtOAc then EtOAc / MeOH) afforded 60g (0.33 g, 51 %).

Data of 60g: C53H58N4O14 (975.0). LC-MS (method 6b): R _t = 2.45 (90%), 975.5 ([M+H] ⁺ ).

Synthesis of the amino acid 61a

A soln of 60a (215 mg, 0.26 mmol) and DMBA (145 mg, 0.92 mmol) in degassed CH2CI2 / EtOAc 1 :1 (8 mL) was treated with Pd(PPh ₃ ) ₄ (30 mg) for 1 h at rt. The volatiles were evaporated. The residue was suspended in Et.20 and filtered, washed (Et ₂ 0) and dried i.v. Purification by FC (CH ₂ CI ₂ / MeOH) gave 61a (163 mg, 89%).

Data of 61a: C ₃₅ H ₄₂ N ₄ 0ii (694.7). LC-MS (method 1 a): R _t = 1 .60 (89%), 695.3 ([M+H] ⁺ ).

Synthesis of the amino acid 61 e

Amino acid 61e (182 mg, 100%; purified by FC (CH2CI2 / MeOH)) was obtained from 60e (210 mg, 0.23 mmol) by following the procedure described for the synthesis of 61a.

Data of 61e: C ₃ 6H ₄ oCIF ₃ N ₄ Oii (797.2). LC-MS (method 1 a): R _t = 1.86 (86%), 797.2 ([M+H] ⁺ ).

Synthesis of the amino acid 61f

Amino acid 61f (129 mg, 84%; purified by FC (CH2CI2 / MeOH)) was obtained from 60f (180 mg, 0.22 mmol) by following the procedure described for the synthesis of 61a.

Data of 61f: C ₃₅ H ₄ iFN ₄ 0n (712.7). LC-MS (method 1 a): R _t = 1 .63 (86%), 713.3 ([M+H] ⁺ ).

Synthesis of the amino acid 61 g

Amino acid 61g (238 mg, 83%; purified by FC (CH2CI2 / MeOH)) was obtained from 60g (330 mg, 0.34 mmol) by following the procedure described for the synthesis of 61a. Data of 61g: C46H50N4O12 (850.9). LC-MS (method 6b): R _t = 1 .64 (91 %), 851.4 ([M+H] ⁺ ).

Synthesis of the macrocyclic ester Ex.3

At 90°C, a soln of 61a (80 mg, 0.1 15 mmol) and i-Pr ₂ NEt (0.1 mL, 0.6 mmol) in dry DMF (40 mL) was added over 2 h to a mixture of 2-chloro-1-methylpyridinium iodide (106 mg, 0.40 mmol) in toluene (500 mL). Partial evaporation of the volatiles, aq. workup (EtOAc, 1 M aq. HCI soln, H2O) and purification of the crude product by FC (hexane / EtOAc / MeOH) afforded Ex.3 (24 mg, 30%).

Data of Ex.3 cf. Table 01a

Synthesis of the macrocyclic ester Ex.4

At 90°C, a soln of 61e (55 mg, 0.069 mmol) and i-Pr ₂ NEt (0.06 mL, 0.35 mmol) in dry DMF (50 mL) was added over 2 h to a mixture of 2-chloro-1-methylpyridinium iodide (64 mg, 0.24 mmol) in toluene (500 mL). Partial evaporation of the volatiles, aq. workup (EtOAc, 1 M aq. HCI soln, H ₂ 0) and purification of the crude product by FC (hexane / EtOAc / MeOH) afforded Ex.4 (13 mg, 23%).

Data of Ex.4 cf. Table 01a

Synthesis of the macrocyclic ester Ex.5

Ester Ex.5 (19 mg, 34%; purified by FC (hexane / EtOAc / MeOH)) was obtained from 61f (58 mg, 0.08 mmol) by applying the procedure outlined for the synthesis of Ex.4. Data of Ex.5 cf. Table 01a Synthesis of the macrocyclic dicarboxylic acid 62a

A soln of Ex.3 (40 mg, 0.06 mmol) in THF / EtOH 4:1 (5 mL) was treated with 2 M aq. LiOH soln (0.075 mL, 0.15 mmol) for 2 h at rt, followed by addn of 1 M aq. HCI soln (5 mL) and extraction with EtOAc. The organic phase was dried (Na ₂ S0 ₄ ), filtered and concentrated. Purification of the crude product by prep. HPLC (method 2a) afforded 62a (15 mg, 41 %).

Data of 62a: C ₃ iH ₃₂ N ₄ Oio (620.6). LC-MS (method 3a): R _t = 1 .01 (93%), 621.1 ([M+H] ⁺ ). ¹ H-NMR (DMSO-de): 13.6 (very br. s, ca 2 H); 7.43 - 7.24 (m, 6 H); 7.17 - 6.98 (br. m, unresolved, 2 H); 6.96 - 6.81 (br. m, unresolved, 1 H); 5.27 (d, J = 12.2, 1 H); 4.87 (d, J = 14.4, 1 H); 4.81 (d, J = 5.8, 1 H); 4.60 (d, J = 9.0, 1 H); 4.28 - 4.10 (m, 4 H); 4.04 - 3.87 (m, 3 H); 3.54 (t-like m, 2 H); ca 3.3 (2 H, superimposed by H ₂ 0 signal); 2.21 - 1 .84 (m, 4 H); 1 .82 - 1.59 (m, 2 H). Synthesis of the macrocyclic dicarboxylic acid 62e

A soln of Ex.4 (25 mg, 0.03 mmol) in THF / EtOH 4:1 (5 mL) was treated with 2 M aq. LiOH soln (0.065 mL, 0.13 mmol) for 2 h at rt, followed by addn of 1 M aq. HCI soln (10 mL) and extraction with EtOAc. The organic phase was dried (Na2S0 ₄ ), filtered and concentrated. Purification of the crude product by prep. HPLC (method 2a) afforded 62e (19 mg, 80%).

Data of 62e: C ₃ 2H3oCIF ₃ N ₄ Oio (723.0). LC-MS (method 1 d): Rt = 1.81 (97%), 723.2 ([M+H] ⁺ ). Synthesis of the macrocyclic dicarboxylic acid 62f

Acid 62f (10 mg, 82%; purified by prep. HPLC (method 2a)) was obtained from Ex.5 (13 mg, 0.019 mmol) by following the procedure described for the synthesis of 62e. Data of 62f: C ₃ iH ₃ iFN ₄ Oio (638.6). LC-MS (method 1 d): Rt = 1 .46 (96%), 639.2 ([M+H] ⁺ ).

Synthesis of the macrocyclic lactam 62g

At 90°C, a soln of 61g (1 19 mg, 0.14 mmol) and i-Pr ₂ NEt (0.12 mL, 0.7 mmol) in dry DMF (45 mL) was added over 2 h to a mixture of 2-chloro-1-methylpyridinium iodide (125 mg, 0.49 mmol) in toluene (1000 mL). Partial evaporation of the volatiles, aq. workup (EtOAc, 1 M aq. HCI soln, H2O) and purification of the crude product by FC (hexane / EtOAc / MeOH) afforded 62g (30 mg, 25%).

Data of 62g: C ₄₆ H ₄₈ N ₄ 0n (832.9). LC-MS (method 6a): R _t = 2.10 (92%), 833.3 ([M+H] ⁺ ). Synthesis of the macrocyclic ester Ex.6

A soln of 62g (62 mg, 0.074 mmol) in EtOH (12.5 mL) was hydrogenated at rt and at normal pressure in the presence of palladium hydroxide on activated charcoal (15- 20% Pd; moistened with 50% H2O; 25 mg) for 5 h. The mixture was filtered through a pad of celite. The filtrate was concentrated and purified by FC (EtOAc) to afford Ex.6 (49 mg, 89%).

Data of Ex.6 cf. Table 01a

Synthesis of the macrocyclic dicarboxylic acid 63b and dicarboxylic acid monoester 63a

A soln of Ex.6 (40 mg, 0.054 mmol) in THF / EtOH 4:1 (5 mL) was treated with 2 M aq. LiOH soln (0.08 mL, 0.16 mmol) for 2 h at rt. More 2 M aq. LiOH soln (0.04 mL, 0.08 mmol) was added and stirring was continued for 2 h followed by the addn of 1 M aq. HCI soln (10 mL). Aq. workup (EtOAc; Na2S0 ₄ ) and purification of the crude product by prep. HPLC (method 2a) afforded 63b (18 mg, 49%) and 63a (6 mg, 16%).

Data of 63a: C37H38N4O11 (714.7). LC-MS (method 3b): R _t = 1 .30 (86%), 715.0 ([M+H] ⁺ ).

Data of 63b: Cssh^NUOn (686.7). LC-MS (method 3b): R _t = 1 .03 (96%), 686.9 ([M+H] ⁺ ). Synthesis of final products Ex.7 and Ex.8 (Scheme 7):

Synthesis of the ester 64

ADDP (3.24 g, 12.8 mmol) in CHCI ₃ (15 mL) was added drop by drop at 0°C to a soln of methyl 4-hydroxybenzoate (1 ; 1.56 g, 10.3 mmol), alcohol 8 (2.7 g, 8.56 mmol) and PPh ₃ (3.37 g, 12.8 mmol) in degassed CHCI3 (35 mL). The mixture was stirred at rt for 16 h followed by aq. workup (CH ₂ CI ₂ , sat. aq. NaHC0 ₃ soln; Na ₂ S0 ₄ ). The resulting crude product was suspended in CH2CI2 / hexane 2:8 and filtered. The filtrate was concentrated and purified by FC (hexane / EtOAc) to afford 64 (3.82 g, 99%).

Data of 64: C23H35NO6S1 (449.6). LC-MS (method 7d): R _t = 9.28 (98%), 450.3 ([M+H] ⁺ ).

Synthesis of the acid 65

At 0°C, LiOH H2O (0.43 g, 10.2 mmol) was added to a soln of 64 (2.3 g, 5.1 mmol) in THF (40 mL), MeOH (10 mL) and H ₂ 0 (10 mL). The mixture was stirred at 0°C to rt for 3 h. More LiOH H2O (0.215 g, 5.2 mmol) was added and stirring was continued for 4 h, followed by a further addn of LiOH H2O (0.1 1 g, 2.6 mmol). The mixture was stirred for addn 2 h, cooled to 0°C, diluted with 1 M aq. HCI soln (50 mL) and extracted with CHCI3. MeOH was added to the organic phase to dissolve precipitating material. The organic phase was dried (Na2S0 ₄ ), filtered and concentrated. Purification of the crude product by FC (hexane / EtOAc then EtOAc / MeOH) gave 65 (1.15 g; 52%).

Data of 65: C22H33NO6S1 (435.6). LC-MS (method 7c): R _t = 9.79 (97%), 436.3 ([M+H] ⁺ ). Synthesis of the amide 66

At 0°C, i-Pr ₂ NEt (1.6 mL, 9.35 mmol) was added to a soln of HATU (1 .28 g, 3.36 mmol), 65 (0.98 g, 2.24 mmol) and 19 CF ₃ C0 ₂ H (80%; 1.2 g, 2.24 mmol) in DMF (30 mL). Stirring was continued at rt for 16 h followed by an aq. workup (EtOAc, sat. aq. NaHCC>3 soln, H2O, sat. aq. NaCI soln; Na2S0 ₄ ) and purification by FC (hexane / EtOAc / MeOH) to afford 66 (1.64 g, ca 80%; used without further purification).

Data of 66: C ₃₆ H ₅₃ N ₃ 0nSi (731 .9). LC-MS (method 1 a): R _t = 2.64 (82%), 732.5 ([M+H] ⁺ ). Synthesis of the carbamate 67

At 0°C, a soln of 22e (0.35 g, 1.19 mmol) and Et ₃ N (0.23 mL, 1 .65 mmol) in dry CH2CI2 (1 1 mL) was slowly added to a soln of phosgene (20% in toluene, 0.87 mL, 1 .66 mmol) in CH2CI2 (1 1 mL). Stirring was continued for 1 h. The volatiles were removed. The residue was suspended in Et.20 and filtered. The filtrate was concentrated to afford crude 26e, which was dissolved in CH2CI2 (1 1 mL) and cooled to 0°C. A soln of 66 (0.8 g, ca 0.9 mmol) and Et ₃ N (0.23 ml, 1.65 mmol) in CH ₂ CI ₂ (1 1 mL) was slowly added, followed by DMAP (0.07 g, 0.57 mmol). The mixture was stirred at rt for 18 h. Silica gel was added to the mixture and the volatiles were evaporated. Purification by FC (hexane / EtOAc then EtOAc / MeOH) afforded 67 (0.495 g, ca 50%; used without further purification).

Data of 67: C ₄₉ H62CIF ₃ N ₄ Oi ₄ Si (1051 .6). LC-MS (method 7d): R _t = 10.25 (83%), 1051.4 ([M+H] ⁺ ).

Synthesis of the amino acid 68

A soln of 67 (980 mg, 0.93 mmol) and DMBA (514 mg, 3.26 mmol) in degassed CH2CI2 / EtOAc 1 :1 (30 mL) was treated with Pd(PPh ₃ ) ₄ (108 mg) for 1 h at rt. The volatiles were evaporated. Purification by FC (CH2CI2 / MeOH) gave 68 (726 mg, 84%).

Data of 68: C ₄₂ H ₅₄ CIF ₃ N ₄ Oi ₂ Si (927.4). LC-MS (method 1 a): R _t = 2.25 (98%), 927.4 ([M+H] ⁺ ).

Synthesis of the macrocyclic lactam 69

At 90°C, a soln of 68 (100 mg, 0.108 mmol) and i-Pr ₂ NEt (0.092 mL, 0.54 mmol) in dry DMF (50 mL) was added over 2 h to a mixture of 2-chloro-1 -methylpyridinium iodide (96 mg, 0.38 mmol) in toluene (1000 mL). Evaporation of the solvents and purification of the residue by FC (hexane / EtOAc / MeOH) afforded 69 (29 mg, 29%). Data of 69: C ₄ 2H52CIF3N ₄ OiiSi (909.4). LC-MS (method 6a): R _t = 2.51 (94%), 909.1 ([M+H] ⁺ ).

Synthesis of macrocyclic diester Ex.7 and dicarboxylic acid 70.

TBAF soln (1 M in THF; 0.235 mL, 0.235 mmol) was added at 0°C to a soln of 69 (140 mg, 0.154 mmol) in THF (16 mL). The mixture was stirred at 0°C for 1 h, followed by an aq. workup (EtOAc, sat. aq. NH ₄ CI soln, H2O, sat. aq. NaCI soln; Na ₂ S0 ) and purification by FC (EtOAc / MeOH) to afford Ex.7 (74 mg, 60%) and 71 (26 mg; used without further purification).

A soln of 71 (16 mg) in THF / EtOH 4:1 (1.25 mL) was treated with 2 M aq. LiOH soln (0.03 mL, 0.06 mmol) for 1 h at rt, followed by addn of 1 M aq. HCI soln and extraction with EtOAc. The organic phase was dried (Na2S0 ₄ ), filtered and concentrated to give 70 (15 mg, 97%).

Data of 70: C ₃ 2H3oCIF ₃ N ₄ Oii (739.0). LC-MS (method 7a): R _t = 5.56 (92%), 739.3 ([M+H] ⁺ ).

Data of Ex.7: cf Table 01a

Synthesis of the alcohol 72

TBAF soln (1 M in THF; 0.87 mL, 0.87 mmol) was added at 0°C to a soln of 64 (325 mg, 0.723 mmol) in THF (30 mL). The mixture was stirred at 0°C for 2.5 h, followed by an aq. workup (EtOAc, sat. aq. NH ₄ CI soln, H2O, sat. aq. NaCI soln; Na2S0 ₄ ) and purification by FC (hexane / EtOAc) to afford 72 (245 mg, 100%).

Data of 72: Ci ₇ H ₂ iN0 ₆ (335.4). ¹ H-NMR (DMSO-d ₆ ): 7.89 (d, J = 8.9, 2 H); 7.05 (d, J = 8.6, 2 H); 5.92 (m, 1 H); 5.27 (qd, J = 1.7, 17.3, 1 H); 5.17 (qd, J = 1 .5, 10.5, 1 H); 5.02 (br. m, 1 H); 4.52 (br. m, 2 H); 4.33 (br. m, 1 H); 4.25 - 4.10 (br. m; 3 H); 3.81 (s, 3 H); 3.38 (br. m, 2 H); 2.10 - 2.00 (br. m, 2 H).

Synthesis of the alcohol 73

NaH dispersion (60% in oil; 88 mg, 2.19 mmol) was slowly added at 0°C to a soln of 72 (245 mg, 0.731 mmol) in DMF (15 mL). The mixture was stirred at rt for 15 min, then heated to 50°C for 1 h and cooled to rt, followed by the addn of Nal (165 mg, 1 .1 mmol) and CH3I (0.91 mL, 14.6 mmol). Stirring was continued for 16 h. Aq. workup (EtOAc, 0.5 M aq. HCI soln; Na ₂ S0 ) and purification by FC (hexane / EtOAc) yielded 73 (201 mg, 79%).

Data of 73: Ci ₈ H ₂ 3N0 ₆ (349.4). LC-MS (method 6b): R _t = 1.64 (95%), 350.2 ([M+H] ⁺ ). Synthesis of the acid 74

At rt, 2 M aq. LiOH soln (1 mL, 2.0 mmol) was added to a soln of 73 (240 mg, 0.69 mmol) in THF (6 mL) and MeOH (1.5 mL). The mixture was stirred for 24 h. More 2 M aq. LiOH soln (0.7 mL, 1 .4 mmol) was added and stirring was continued for 24 h. The mixture was diluted with 1 M aq. HCI soln (50 mL) and extracted with EtOAc. The organic phase was dried (Na ₂ S0 ₄ ), filtered and concentrated to give 74 (225 mg, 98%).

Data of 74: Ci ₇ H ₂ iN0 ₆ (335.4). LC-MS (method 6b): R _t = 1.31 (90%), 336.2 ([M+H] ⁺ ). Synthesis of the amide 75

At 0°C, i-Pr ₂ NEt (0.55 mL, 3.2 mmol) was added to a soln of HATU (0.408 g, 1 .07 mmol), 74 (0.24 g, 0.716 mmol) and 19 CF ₃ C0 ₂ H (80%; 0.383 g, 0.716 mmol) in DMF (6 mL). Stirring was continued at rt for 3 h followed by an aq. workup (EtOAc, 1 M aq. HCI soln, sat. aq. NaHCC>3 soln, H ₂ 0, sat. aq. NaCI soln; Na ₂ S0 ₄ ) and purification by FC (hexane / EtOAc then EtOAc / MeOH) to afford 75 (0.38 g, 84%).

Data of 75: C ₃ iH ₄ i N ₃ 0n (631 .7). LC-MS (method 6b): R _t = 1.44 (89%), 632.2 ([M+H] ⁺ ).

Synthesis of the carbamate 76

At 0°C, a soln of 22e (0.21 g, 0.66 mmol) and Et ₃ N (0.15 mL, 1 .07 mmol) in dry CH ₂ CI ₂ (3 mL) was slowly added to a soln of phosgene (20% in toluene, 0.48 mL, 0.9 mmol) in CH ₂ CI ₂ (3 mL). Stirring was continued for 1 h. The volatiles were removed. The residue was suspended in Et ₂ 0 and filtered. The filtrate was concentrated to afford crude 26e, which was dissolved in CH ₂ CI ₂ (3 mL) and cooled to 0°C. A soln of 75 (0.38 g, 0.6 mmol) and Et ₃ N (0.15 ml, 1 .07 mmol) in CH ₂ CI ₂ (3 mL) was slowly added, followed by DMAP (0.037 g, 0.30 mmol). The mixture was stirred at rt for 18 h. Silica gel was added to the mixture and the volatiles were evaporated. Purification by FC (hexane / EtOAc then EtOAc / MeOH) afforded 76 (0.227 g, 40%).

Data of 76: C ₄₄ H ₅₀ CIF ₃ N ₄ Oi ₄ (951 .3). LC-MS (method 6b): R _t = 2.21 (87%), 951.3 ([M+H] ⁺ ).

Synthesis of the amino acid 77

A soln of 76 (224 mg, 0.24 mmol) and DMBA (130 mg, 0.82 mmol) in degassed CH ₂ CI ₂ / EtOAc 1 :1 (8 mL) was treated with Pd(PPh ₃ ) ₄ (28 mg) for 1 h at rt. The volatiles were evaporated. Purification by FC (CH ₂ CI ₂ / MeOH) gave 77 (185 mg, 95%). Data of 77: C37H42CIF3N4O12 (827.2). LC-MS (method 6b): R _t = 1.51 (93%), 827.2 ([M+H] ⁺ ).

Synthesis of the macrocyclic ester Ex.8

At 85°C, a soln of 77 (50 mg, 0.06 mmol) and i-Pr ₂ NEt (0.052 mL, 0.30 mmol) in dry DMF (25 mL) was added over 2 h to a mixture of 2-chloro-1-methylpyridinium iodide (54 mg, 0.21 mmol) in toluene (500 mL). Evaporation of the solvents and purification of the residue by FC (hexane / EtOAc / MeOH) afforded Ex.8 (10 mg, 21 %).

Data of Ex.8: cf Table 01a

Synthesis of the macrocyclic dicarboxylic acid 78

A soln of Ex.8 (24 mg, 0.03 mmol) in THF / EtOH 4:1 (2.5 mL) was treated with 2 M aq. LiOH soln (0.03 mL, 0.06 mmol) for 7 h at 0°C to rt. More 2 M aq. LiOH soln (0.02 mL, 0.04 mmol) was added and stirring continued for 2 h, followed by addn of 1 M aq. HCI soln (0.1 ml) and evaporation of the volatiles. The residue was suspended in EtOAc and filtered. The filtrate was concentrated to give 78 (22 mg, 98%).

Data of 78: C33H32CIF3N4O11 (753.1 ). LC-MS (method 1 d): Rt = 1.78 (85%), 753.2 ([M+H] ⁺ ). Synthesis of macrocyclic esters 89 and 90 (Scheme 8):

Synthesis of the ester 79 and acid 81

ADDP (4.41 g, 17.46 mmol) was added to a soln of methyl 4-hydroxybenzoate (1 ; 2.125 g, 14.0 mmol), alcohol 11 (2.32 g, 1 1 .6 mmol) and PPh ₃ (4.58 g, 17.46 mmol) in degassed CHCI3 (50 mL). The mixture was diluted with CHCI3 (20 mL) and stirred at rt for 20 h followed by evaporation of the volatiles. The resulting crude product was purified by FC (hexane / EtOAc) to afford pure 79 (0.36 g, 9%) and 79 (1 .7 g, ca 35%, purity ca 80%; used without further purification).

At 0°C, 2 M aq. LiOH soln (4 mL, 8 mmol) was added to a soln of 79 (1.67 g) in THF / MeOH 3:1 (16 mL). The mixture was stirred at 0°C to rt for 16 h, cooled to 0°C and acidified by the addn of 2 M aq. HCI soln (8 mL). The mixture was extracted with EtOAc. The organic phase was dried (Na2S0 ₄ ), filtered, concentrated and purified by FC (hexane / EtOAc / MeOH) to afford 81 (1 .18 g, 92%).

Data of 79: C18H23NO5 (333.4). LC-MS (method 8a): R _t = 1.62 (95%), 334.1 ([M+H] ⁺ ). Data of 81 : C17H21 NO5 (319.4). LC-MS (method 1 a): R _t = 1.98 (80%), 320.2 ([M+H] ⁺ ). Synthesis of the ester 80 and acid 82

Ester 80 (2.87 g, 96%, purified by FC (hexane / EtOAc)) was obtained from methyl 4- hydroxybenzoate (1 ; 1 .63 g, 10.7 mmol), alcohol 12 (1.8 g, 8.95 mmol) by applying the procedure outlined for the synthesis of 79.

Acid 82 (2.51 g, 95%) was obtained from 80 (2.76 g, 8.25 mmol) by following the procedure described for the synthesis of 81.

Data of 80: Ci ₇ H ₂ iN0 ₆ (335.4). LC-MS (method 1 a): R _t = 2.05 (98%), 336.1 ([M+H] ⁺ ). Data of 82: Ci ₆ Hi ₉ N0 ₆ (321.3). LC-MS (method 1 a): R _t = 1.67 (97%), 322.1 ([M+H] ⁺ ). Synthesis of the amide 83

To a soln of 81 (0.68 g, 1 .92 mmol) and 19 CF ₃ C0 ₂ H (0.75 g, 1.75 mmol) in DMF (35 mL) was added a soln of HATU (0.43 M in DMF; 5.3 mL, 2.28 mmol) and then slowly at 0°C i-Pr ₂ NEt (0.9 mL, 5.26 mmol). Stirring was continued at 0°C for 2 h followed by an aq. workup (EtOAc, sat. aq. NaHCC>3 soln; Na ₂ S0 ₄ ) and purification by FC (hexane / EtOAc / MeOH) to afford 83 (0.8 g, 74%; used without further purification). Data of 83: C ₃ i H ₄ iN ₃ Oio (615.7). LC-MS (method 2c): R _t = 2.02 (85%), 616.3 ([M+H] ⁺ ).

Synthesis of the amide 84

To a soln of 82 (0.66 g, 2.05 mmol) and 19 CF ₃ C0 ₂ H (0.8 g, 1 .64 mmol) and HATU (0.937 g, 2.47 mmol) in DMF (36 mL) was slowly added at 0°C i-Pr ₂ NEt (1.3 mL, 7.59 mmol). Stirring was continued at 0°C for 2 h followed by an aq. workup (EtOAc, sat. aq. NaHC0 ₃ soln; Na ₂ S0 ₄ ) and purification by FC (hexane / EtOAc / MeOH) to afford 84 (0.53 g, 52%).

Data of 84: C ₃₀ H ₃₉ N ₃ Oii (617.6). LC-MS (method 1 a): R _t = 1.80 (96%), 618.2 ([M+H] ⁺ ).

Synthesis of the carbamate 85

At 0°C, a soln of 22e (0.228 g, 0.78 mmol) and Et ₃ N (0.32 mL, 2.3 mmol) in dry CH ₂ CI ₂ (4 mL) was slowly added to a soln of phosgene (20% in toluene, 0.56 mL, 1 .06 mmol) in CH ₂ CI ₂ (4 mL). Stirring was continued for 1 h. The volatiles were removed. The residue was suspended in Et ₂ 0 and filtered. The filtrate was concentrated to afford crude 26e, which was dissolved in CH ₂ CI ₂ (4 mL) and cooled to 0°C. A soln of 83 (0.386 g, 0.62 mmol) and Et ₃ N (0.32 ml, 2.3 mmol) in CH ₂ CI ₂ (4 mL) was slowly added, followed by DMAP (0.033 g, 0.275 mmol). The mixture was stirred at rt for 16 h. Silica gel was added to the mixture and the volatiles were evaporated. Purification by FC (hexane / EtOAc then EtOAc / MeOH) afforded 85 (0.186 g, 30%).

Data of 85: C ₄ 4H ₅ oCIF ₃ N40i3 (935.3). LC-MS (method 6b): R _t = 2.35 (85%), 935.4 ([M+H] ⁺ ).

Synthesis of the amino acid 87

A soln of 85 (299 mg, 0.32 mmol) and DMBA (126 mg, 0.80 mmol) in degassed CH2CI2 / EtOAc 1 :1 (6 mL) was treated with Pd(PPh ₃ ) ₄ (37 mg) for 4 h at rt. The volatiles were evaporated. Purification by FC (CH2CI2 / MeOH) gave 87 (205 mg, 79%).

Data of 87: C37H ₄₂ CIF ₃ N ₄ Oii (81 1 .2). LC-MS (method 6b): R _t = 1.53 (87%), 81 1.3 ([M+H] ⁺ ).

Synthesis of the carbamate 86 and of the amino acid 88

Carbamate 86 (179 mg, 36%, purification by FC (hexane / EtOAc then EtOAc / MeOH) was obtained from 84 (332 mg, 0.538 mmol) and 22e (200 mg, 0.681 mmol) applying the procedure outlined for the synthesis of 85.

A soln of 86 (172 mg, 0.18 mmol) and DMBA (87 mg, 0.55 mmol) in degassed CH2CI2 / EtOAc 1 :1 (8 mL) was treated with Pd(PPh ₃ ) ₄ (40 mg) for 4 h at rt. The volatiles were evaporated. Purification by FC (CH2CI2 / MeOH) gave 88 (152 mg; 100%).

Data of 88: C ₃ 6H ₄ oCIF ₃ N ₄ Oi2 (813.2). LC-MS (method 1 a): R _t = 1.84 (97%), 813.4 ([M+H] ⁺ ). Synthesis of the macrocyclic ester 89 and acid 91

At 90°C, a soln of 87 (54 mg, 0.066 mmol) and i-Pr ₂ NEt (0.033 mL, 0.19 mmol) in dry MeCN (50 mL) was added over 2 h to a mixture of 2-chloro-1-methylpyridinium iodide (35 mg, 0.14 mmol) in MeCN (1000 mL). Stirring at 90°C was continued for 1 h. Evaporation of the solvents and purification of the residue by FC (hexane / EtOAc / MeOH) afforded 89 (1 1 mg, 21 %).

A soln of 89 (7.3 mg, 0.009 mmol) in THF (0.8 mL) was treated with 0.58 M aq. LiOH soln (0.05 mL, 0.029 mmol) for 0.5 h at 0°C to rt. More 0.58 M aq. LiOH soln (0.05 mL, 0.029 mmol) was added and stirring continued for 0.5 h, followed by addn of 3 M aq. HCI soln (0.05 ml) and evaporation of the volatiles. The residue was purified by prep. HPLC (method 2a) to give the macrocyclic diacid 91 (1 .6 mg, 23%). Data of 91 : C33H32CIF3N4O10 (737.1 ). LC-MS (method 6a): R _t = 1 .49 (97%), 736.9 ([M+H] ⁺ ).

Synthesis of the macrocyclic ester 90 and acid 92

The macrocyclic diester 90 (7 mg, 12%) was obtained from 88 (62 mg, 0.076 mmol) by following the procedure outlined for the synthesis of 89.

Macrocyclic diacid 92 (0.7 mg, 16%, purification by prep. HPLC (method 2a)) was obtained from 90 (4.6 mg) by following the procedure outlined for the synthesis of macrocyclic diacid 91 .

Data of 92: C32H30CIF3N4O11 (739.0). LC-MS (method 6a): R _t = 1 .36 (97%), 739.0 ([M+H] ⁺ ).

Table 01a: Analytical data of Examples (Ex.)

Monoisotopic [M+H] ⁺

Ex. Formula Rt (purity) LC-MS

Mass found

Ex.1 C43H42CI2F3N5O11 931 .22 2.39 (95%) 932.3 method 1 d

Ex.2 C43H43CIF3N5O11 897.26 1 .90 (96%) 898.3 method 6a

Ex.3 C35H40N4O10 676.27 2.03 (91 %) 677.3 method 1 d

Ex.4 C36H38CIF3N4O10 778.22 2.34 (87%) 779.2 method 1 d

Ex.5 C35H39FN4O10 694.27 2.06 (87%) 695.3 method 1 d

Ex.6 C39H42N4O11 742.29 1 .61 (92%) 743.2 method 6a

Ex.7 C36H38CIF3N4O11 794.22 2.05 (93%) 795.3 method 1 d

Ex.8 C37H40CIF3N4O11 808.23 2.28 (93%) 809.3 method 1 d Table 01 b: lUPAC names of Examples (Ex.)

Ex. lUPAC name

diethyl 1 -{[(65,14 ?,165,245)-14-[(3-chlorobenzoyl)amino]-9-[4-chloro-3- (trifluoromethyl)phenyl]-2,8,1 1 -trioxo-7,18-dioxa-3,9,12-

Ex.1

triazatetracyclo[17.2.2.1 ^{3 6} .0 ^{12 16} ]tetracosa-1 (21 ),19,22-trien-24- yl]carbonyl}-3,3-azetidinedicarboxylate

diethyl 1 -{[(65,14 ?,165,245)-14-[(3-chlorobenzoyl)amino]-2,8,1 1-trioxo- 9-[3-(trifluoromethyl)phenyl]-7, 18-dioxa-3,9, 12-

Ex.2

triazatetracyclo[17.2.2.1 ^{3 6} .0 ¹² ' ¹⁶ ]tetracosa-1 (21 ),19,22-trien-24- yl]carbonyl}-3,3-azetidinedicarboxylate

diethyl 1 -{[(6 S, 165,24S)-2,8, 1 1 -trioxo-9-phenyl-7, 18-dioxa-3,9, 12-

Ex.3 triazatetracyclo[17.2.2.1 ^{3 6} .0 ¹² ' ¹⁶ ]tetracosa-1 (21 ),19,22-trien-24- yl]carbonyl}-3,3-azetidinedicarboxylate

diethyl 1 -{[(65,165,245)-9-[4-chloro-3-(trifluoromethyl)phenyl]-2,8,1 1-

Ex.4 trioxo-7,18-dioxa-3,9,12-triazatetracyclo[17.2.2.1 ^{3 6} .0 ^{12 16} ]tetracosa-

1 (21 ),19,22-trien-24-yl]carbonyl}-3,3-azetidinedicarboxylate diethyl 1 -{[(6 S, 165,245)-9-(4-fluorophenyl)-2,8, 1 1 -trioxo-7, 18-dioxa-

Ex.5 3,9,12-triazatetracyclo[17.2.2.1 ^{3 6} .0 ¹² ' ¹⁶ ]tetracosa-1 (21 ), 19,22-trien-24- yl]carbonyl}-3,3-azetidinedicarboxylate

diethyl 1 -{[(65,165,245)-9-(5-hydroxy-2-naphthyl)-2,8,1 1 -trioxo-7, 18-

Ex.6 dioxa-3,9,12-triazatetracyclo[17.2.2.1 ^{3 6} .0 ^{12 16} ]tetracosa-1 (21 ), 19,22- trien-24-yl]carbonyl}-3,3-azetidinedicarboxylate

diethyl 1 -{[(65,14 ?,165,245)-9-[4-chloro-3-(trifluoromethyl)phenyl]-14- hyd roxy-2 , 8 , 1 1 -tri oxo-7 , 18-d ioxa-3 , 9 , 12-

Ex.7

triazatetracyclo[17.2.2.1 ^{3 6} .0 ¹² ' ¹⁶ ]tetracosa-1 (21 ),19,22-trien-24- yl]carbonyl}-3,3-azetidinedicarboxylate

diethyl 1 -{[(65,14 ?,165,245)-9-[4-chloro-3-(trifluoromethyl)phenyl]-14- methoxy-2,8, 1 1 -trioxo-7, 18-dioxa-3,9, 12-

Ex.8

triazatetracyclo[17.2.2.1 ^{3 6} .0 ^{12 16} ]tetracosa-1 (21 ),19,22-trien-24- yl]carbonyl}-3,3-azetidinedicarboxylate Scheme 2 loc DMS

4 R ⁱ x = H 6 R ¹ " = Cbz

b)

5 R ^IX = TBDMS 7 R ¹ " = H

8 R ^m = Alloc a) TBDMSCI, imidazole, DMF

b) H ₂ , Pd-C, MeOH

c) Allyl chloroformate, CH ₂ CI ₂ , aq. NaHC0 ₃ soln

9 X = CH ₂ 11 X = CH ₂

10 X = O 12 X = 0

Scheme 2, continued

Allyl chloroacetate

,NH,

Ar' NL Ο,ΑΙΜ nBu ₄ NI, DMF (THF) Ar

21a -21h 22a - 22h

21a, 22a Ar :

21 d, 22d Ar:

C ₆ H ₅ CH ₂ OH,

Scheme 3 BooHN

ADDP, PPh , CHC

THF, CH OH

H COCI ₂ °Y ^CI NEt ₃ , ,N^C0 ₂ Allyl ^N^CO^IIyl DMAP,

Et,N, CH,CI CH2CI2

22a - 22c 26a - 26c

Bo

27a - 27c R" = Allyl, R ^IM = Pd(PPh ₃ ) ₄ , 29a - 29c

1 ,3-dimethylbarbituric acid,

28a - 28c R" = H, R ¹ " = H CH ₂ CI ₂ , EtOAc Scheme 3, continued

29a - 29c R ^lv = Boc V :

TFA, CH ₂ CI ₂ LiOH, H ₂ 0, (HCI-dioxane for 29c) THF, CH ₃ OH

30a - 30c R' ^v = H 32a - 32c R V ^v = . H

Scheme 4

2

3

43d - 43e

43d - 43e R" = Allyl, R ¹ " = Alloc Pd(PPh ₃ ) ₄ , 45d - 45e R ^v = CH

1 ,3-dimethylbarbituric acid, LiOH, H ₂ 0,

THF, CH3OH 44d - 44e R ¹¹ = H, R ¹ " = H CH ₂ CI ₂ , EtOAc 46d - 46e R v . Scheme 4, continued

Scheme 4, continued

52a n = 1 , R ^VI = H

52b n = 0, R ^VI =H

c _\ " X. ' ^tetra butylammonium iodide, i-Pr ₂ NEt, CH ₃ CN b) , Et ₃ N, W-methylpyrrolidone

1) Reaction conditions, for example, as described for the preparation of Example 100 in WO2004/087720 A1 (for a) and for the preparation of Example 50 in WO2009/069100 A1 (for b).

Scheme 5

54e, 54h R" = Allyl, R

55e, 55h R" = H, R ^m =

a) Pd(PPh ₃ ) ₄ , 1 ,3-dimethylbarbituric acid, CH ₂ CI ₂ , EtOAc b) LiOH, H ₂ 0, THF, EtOH Scheme 6

60a, 60e-60g R" = Allyl, R ¹ " Ex.3, Ex.4, Ex.5 R ^vl = Et/Et 61a, 61e-61g R" = H, R ^m = 62a, 62e, 62f R ^vl = H/H

a) Pd(PPh ₃ ) ₄ , 1,3-dimethylbarbituric acid, CH ₂ CI ₂ , EtOAc

b) LiOH, H ₂ 0, THF, EtOH

c) H ₂ , Pd(OH) ₂ -C, EtOH Scheme 7

67

a) Pd(PPh ₃ ) ₄ , 1,3-dimet ylbarbituricacid, CH ₂ CI ₂ , EtOAc

b) TBAF, THF

c) LiOH, H ₂ 0, THF, EtOH Scheme 7, continued

76 R" = Allyl, R ^m = Alloc

Ex.8 R ^VI = Et/Et

g)

77 R" = H, R ^m = H → 78 R ^VI = HI H

Scheme 8

a

85 X = CH ₂ 86 X = 0

85 X = CH ₂ , R" = Allyl, R ^MI = Alloc 89 X = CH ₂ , R ^VI = Et 86 X = 0, R" = Allyl, R ^MI = Alloc b) 90 X = O, R ^VI = Et

87 X = CH ₂ , R" = H, R'" = H c)

88 X = O, R" = H, R'" = H b) 91 X = CH ₂ , R ^vl = H

92 X = O, R ^VI = H b) Pd(PP ₃ ) ₄ , 1 ,3-dimethylbarbituric acid, CH ₂ CI ₂ , EtOAc

c) LiOH, H ₂ 0, THF Biological methods Preparation of the compounds

Compounds were weighed on a microbalance (Mettler MX5) and dissolved in 100% DMSO to a final concentration of 10 mM unless otherwise stated. Stock solutions were kept at +4°C, and protected from light.

Expression and purification of human R14A mutant Pin1

E.coli BL21 pLysE(DE3) cells (New England BioLabs, Ipswich, USA) were transformed with pET14b vector (Merck Millipore, Darmstadt, Germany) encoding for full length Pin1 (amino acids 1-163) with a stabilizing mutation in position 14 (arginine to alanine replacement; Y. Zhang et a/., ACS Chem Biol. 2007, 2(5), 320-328), a N- terminal 6xHis tag and a thrombin cleavage site, and were inoculated into 50 mL of lysogeny broth medium (LB medium; per liter: 10 g tryptone, 5 g yeast extract, 10 g NaCI), containing 100 μg/mL ampicillin and 34 μg/mL choramphenicol. The overnight culture was diluted 100-fold in LB medium containing 100 μg/mL ampicillin and 34 μg/mL choramphenicol. The diluted culture was shaken at 150 rpm and 37°C to an optical density at 595 nm (OD595) of 0.8. Subsequently, 0.25 mM isopropyl-3-D- thiogalactopyranosid (IPTG) was added and the culture was shaken for 4 hours at 150 rpm and 37°C. The cell culture was centrifuged at 5000 rcf for 20 minutes. The pellets were resuspended in 10x buffer A (25 mM Tris-HCI (pH 8.0), 0.5 M NaCI, 10 mM imidazole, 10 mM 2-mercaptoethanol and 1 % Tween 20 ( ν/\ή). The suspension was passed through a high-pressure microfluidizer. The homogenate was centrifuged down in a Sorvall centrifuge using an SS-34 rotor at 9000 rcf and 4°C for 30 minutes. The clear supernatant was kept for further purification.

The clarified supernatant was loaded onto a 5 mL Ni-NTA column at 2.5 mL/minute. The column was washed with 50 mL buffer A. A step gradient was applied at 2.5 mL/minute from 100% buffer A to 100 % buffer B (25 mM Tris-HCI (pH 8.0), 0.5 M NaCI, 10 mM 2-mercaptoethanol and 1 % Tween 20 { ν/\ή, 500 mM imidazole). Fractions of 5 mL were collected and analyzed by SDS-PAGE (4-12%). The fractions containing 6xHis Pin1 were collected and pooled. Pooled His-tagged Pin1 was digested with thrombin (1 U thrombin/mg protein; Sigma-Aldrich) during dialysis for 16 hours at 4°C (molecular weight cut-off: 3 kDa; dialysis buffer: 50 mM Tris-HCI (pH 8.0), 0.15 M NaCI, 5 mM MgCI ₂ , 2.5 mM CaCI ₂ , 1 mM DTT, 10% glycerol { ν/ή).

The overnight solution was passed through a Ni-NTA column (5 mL). The flow through was collected and concentrated to a volume of 2 mL for final size exclusion chromatography. The concentrate was loaded on a Superdex 75 column (90 ml.) with 2 mL/minute and eluted with buffer S (10 mM Hepes (pH 7.5), 0.1 M NaCI, 1 mM DTT). The fractions containing monomeric Pin1 were collected and pooled. Pooled Pin1 was concentrated to about 20 mg/mL for further studies (see below).

Pin1 Fluorescence polarization binding assay

Compounds were tested for competitive binding, using fluorescein-labeled Pintide (fluorescein-5(6)-carboxamidocaproyl-Trp-Phe-Tyr-Ser(P(0)(OH )2)-Pro-Phe-Leu-Glu; P.-J. Lu et a/., Science 1999, 283, 1325; WO 2004/005315 A2) and human mutant R14A Pin1 protein (see above). Serial dilutions of compounds were prepared in 100% DMSO and further diluted in assay buffer (25 mM MOPS, pH 7.5) to reach 0.5% final DMSO concentration ( ι/Λ). Compound dilutions were transfered to black polystyrene non-binding surface NBS 384-well microtiterplate (Corning, Cat. No. 3575). 300 nM of Pin1 was pre-incubated with compounds for 15 minutes, at room temperature, with gentle shaking. Finally, 25 nM of fluorescein-labeled Pintide was added and reaction incubated 15 minutes at room temperature with gentle shaking at 300 rpm, in the dark. Fluorescence polarization was subsequently measured on a Victor2 reader (Perkin Elmer) at room temperature, with an excitation wavelength at 485 nm and emission at 535 nm. The dose-response data were fitted to the 4- parameter Hill equation providing the I C50 value using Graphpad (Prism 5). Results are depicted in Table 02 below. I C50 values are given as mean (where applicable), further indicating the standard deviation (SD) and number of experiments (n).

Cell viability assay (THP-1)

Compounds were tested in a cell viability assay on THP-1 cells (obtained from ATCC). The cell line was grown as recommended by the supplier. THP-1 cells were then seeded on 96-well plate with RPMI 1640 medium (Sigma Aldrich) containing 10% fetal bovine serum ( 1 /1/) (Gibco), 0.05 mM 2-mercaptoethanol and 100 U^g penicillin and streptomycin (Thermo Fisher Scientific), at a density of 300Ό00 cells/mL in 75 uL. Serial dilutions of compounds were added to the cells (25 uL/well, 0.5% DMSO final, v/V) and were incubated at 37°C and 5% C0 ₂ for 72 hours. After incubation, 20 μΙ_ of CellTiter-Blue reagent (Promega) was added to each well. The plate was further incubated at 37°C and 5% CO2 for 1.5 hours. The fluorescence signal of each sample was measured using a Flexstation fluorescence reader with excitation at 560 nm and emission at 590 nm. The experiments were performed in duplicate. Results are depicted in Table 03 below. Table 02: IC50 values from the Pin1 fluorescence polarization binding assay ^*)

1 ) Diacid 56e obtained from hydrolysis of Ex.1

2) Diacid 56h obtained from hydrolysis of Ex.2

3) Diacid 62a obtained from hydrolysis of Ex.3

4) Diacid 62e obtained from hydrolysis of Ex.4

5) Diacid 62f obtained from hydrolysis of Ex.5

6) Diacid 63b obtained from hydrolysis of Ex.6

7) Diacid 70 obtained from hydrolysis of Ex.7

8) Diacid 78 obtained from hydrolysis of Ex.8

n.a. = not applicable

^* ) Ex.1 - Ex.8 are inactive on Pin1 and comprise at least one ester moiety in substituent E, as described herein above. Respective acids obtained from hydrolysis of Ex.1 - Ex.8 show activity on Pin1. Table 03: IC50 values from the cell viability assay on THP-1 cells ^*)

*) IC50 values given as mean, further indicating the

standard deviation (SD) and number of experiments (n)

Proliferation assay (MHHES1 , A673, and BT20)

Ex.7 was tested in proliferation assays on MHHES1 cells (obtained from DSMZ cell line collection), A673 cells (obtained from ECACC cell line collection), and BT20 cells (obtained from ATTC cell line collections) applying the methodology described by E. Prinz et al. (Supplementary Information S18-S21. In: J. Med. Chem. 2011 , 54, 4247- 4263). Briefly, the cell lines were grown as recommended by the suppliers. Cells were then treated for 72 hours with Ex.7 at a maximum concentration of 25 μΜ and in serial 10-fold dilutions. Subsequently, cell proliferation was determined by means of sulforhodamine B staining, and GI50 values were calculated.

Results are depicted in Table 04 below.

Table 04: GI50 values of Ex.7

Cell line Gl ₅₀ [μΜ]

MHHES1 6.1

A673 7.5

BT20 13.0