The present invention relates to compounds active as modulators (inhibitors or activators) of the proteasome activity in mammals, including man, to their process for their preparation, and to their uses for the treatment of pathologies involving the proteasome.

The ubiquitin-proteasome system is the major pathway of proteolysis in eukaryotic cells (Ciechanover, A. EMBO J, 1998, 17, 7151-7160). The eukaryotic proteasome 26S (2.4 MDa) is a multicatalytic protease consisting of a 2OS proteolytic core particle and a 19S regulatory subunit at either or both ends (Groll, M.; Huber, R.

Int. J. Biochem. Cell Biol. 2003, 35, 606). The multifunctional complex is composed of at least 44 polypeptides and has unique properties. Among them, we can point out the 6 active sites (two of chyrnotryptic-, two of tryptic- and two of caspase-like activities) which are segregated in a secluded compartment which favours a processive degradation of proteins. Proteasome is also a N-terminal threonine hydrolase.

Proteasome 26S recognizes polyubiquinated protein and is ATP-dependent. Mammalian cells contain another regulatory complex that associates with the 2OS proteasome: the I IS regulator or PA28 which promotes the production of antigenic peptides. The 20S proteasome degrades oxidized proteins (Davies K. J. A. Biochimie, 2001 , 301-310) and also an increasing amount of non ubiquitinylated proteins such as the proto-oncogenic c-Fos protein (Bossis G., Frerrar P., Acquaviva C, Jariel-Encontre L, Piechaczyk M. MoI. Cell. Biol. 2003, 23, 7425-7436).

Proteasomes are found in both the nucleus and the cytoplasm of eukaryotic cells, hi the cytoplasm, proteasomes localize near centrosomes, on the outer surface of the endoplasmic reticulum and in cytoskeletal networks. hi addition to removing of damaged and unneeded proteins, proteasome degrades key regulatory proteins, which are crucial for many intracellular processes, including cell progression, apoptosis, NF-κB activation and antigen presentation (Coux, O. ; Tanaka, K. ; Goldberg, A. L. Annu. Rev. Biochem. 1996, 65, 801-847; Ciechanover, A. EMBO J., 1998, 17, 7151-7160). Many proteasome substrates are known mediators of pathways that are dysregulated with neoplasia (Adams J. Cancer- Cell 2003, 5, 417- 421 ; Adams J. Nature Reviews/Cancer 2004, 4, 349-359). Proteasome affects cell- cycle progression by regulating the cyclins, and increasing or decreasing the apoptotic activity through effects on caspases, Bcl2 activity and nuclear factor NF-κB.

Remarkably, an empirical finding is that malignant cells are more susceptible to certain proteasome inhibitors than normal cells: reversal or bypass of some of the effects of the mutations in cell-cycle and apoptotic checkpoints that have led to tumorigenesis; higher dependency of malignant cells to proteasome system to remove aberrant proteins, dependence of some tumors to maintain drug or radiation resistance

(Adams J. Nature Reviews/Cancer 2004, 4, 349-359 ; Boccadoro M., Morgan G., Canevagh J. ) Cancer Cell Intern. 2005, 5:18; doi:10. .1186/1475-2867-5-18).

Among natural and synthetic proteasome inhibitors (their structures are precisely described below), only two compounds are in clinical development : Velcade® (bortezomib, or PS341) in cancer and PS-519 in inflammation. In addition to direct apoptotic effects, proteasome inhibitors are reported to enhance sensitivity to standard chemotherapy, radiationjherapy or immunotherapy, and to overcome drug resistance. NF-κB is activated by radiotherapy and chemotherapy in malignant tissues and proteasome inhibition blocks NF-κB activation by preventing proteasomal degradation of I KB (Cusak. Jr et al. . Cancer Res. , 2001, 61, 3535-3540).

PS-341 or bortezomib or Velcade® PS-519

Bortezomib is the first proteasome inhibitor to be approved for the treatment of multiple myeloma based on several types of data : direct inhibition of cancer cells, interference with the adhesion of multiple myeloma cells to bone marrow stroma cells and with production 11-6 in the bone marrow, anti-angiogenic properties (Adams J.

Cancer Cell 2003, 5, 417-421 ).

Bortezomib is administered as cyclical therapy (twice-weekly treatment for 2 weeks every 3 weeks). Proteasome activity is maximally inhibited over 1 h after dosing (Orlowski RZ et al. J. CHn. Oncol. 2002, 20, 4420-4427). Adverse events have been reported in 30% patients enrolled in clinical trials (thrombocytopaenia, fatigue, peripheral neuropathy and neutropenia). Trials are in progress to investigate the use of bortezomib alone or in numerous combinations in order to evaluate its therapeutic value in various cancers (solid and liquid tumors). Results on non-Hodgin's lyphoma, colorectal, lung, breast and prostate cancers appear to be encouraging.

A rather limited number of proteasome inhibitors have been reported (Kisselev A. F., Goldberg A. L. Chemistry & Biology, 2001, 8, 739-758; Reboud-Ravaux M (2002) " Proteasome inhibitors " in Protein Degradation in Health and Diseases, M. Reboud- Ravaux (Ed.), Progress in Molecular and Subcellular Biology, Springer-Verlag, Berlin, Heidelberg, New York; Papapostolou D., Reboud-Ravaux M. J, 5Oc. Biol, 2004, 198,

263-278). Most are short peptides linked at the C-terminus to a reactive group (figure 1) which binds to the catalytic O ^γ -Thrl of the 6 catalytic sites of the proteasome with formation of a reversible (peptide aldehydes), poorly reversible (peptide boronates), or irreversible covalent adduct (peptide vinyl sulfones, peptide epoxyketones)(figure 2A and 2B).

The natural product lactacystin is a non peptidic molecule which cannot penetrate the cells (Figure 2B). At neutral pH, it is rapidly hydrolyzed to give β-lactone which easily enters cells. The β-lactone reacts with O ^Y -Thrl to give a stable covalent acyl- enzyme (ty ₂ = 20 h). Lactacystin does not react specifically with proteasome since cathepsin A, a lysosomal carboxypeptidase and cytosolic tripeptidyl peptidase II are also inhibited. The natural epoxyketones (epoxomicin and eponemicin) have the unique particularity to react with O ^γ and Ot-NH ₂ of Thrl. This probably explains that these compounds are the most selective proteasome inhibitors but they irreversibly inhibit proteasome. Several polyol compounds such as (-)epigallocatechin-3-gallate give stable acyl-enzymes upon reaction with proteasomes.

Non covalent inhibitors have been investigated less extensively, and in principle, should lower side-effects. Only three classes of such inhibitors are known. Ritonavir (Schmidtke, G.; Holzhϋtter, H.-G.; Bogyo, M.; Kairies, N.; Groll, M.; De Giuli, R.; Emch, S.; Groettrup, M. J Biol. Chem. 1999, 274, 35734-35740) and benzylstatine derivatives (Furet, P.; Imbach, P.; Noorani, M.; Koeppler J.; Laumen, K.; Lang, M.;

Guagnano, P. F.; Roesel, J.; Zimmermann, J.; Garcia- Echeverria, C. J. Med. Chem. 2004, 47 (20), 4810 - 4813.; Furet, P.; Garcia- Echeverria, C; Imbach, P.; Lang, M.; Zimmermann, J. (Novartis) PCT Int. AppL, WO 2001089282; 2001.) were shown to inhibit proteasome non covalently (figure 3). A cyclic peptide TMC-95A which is a metabolite of Apiospora montagnei is a potent reversible inhibitor with no inhibition of m-calpain, cathepsin-L and trypsin (Onuki, T.; Sugita, N., Kono, O.; Kogushi, Y.; Murakami, T.; Nishio, M., (Tanabe Seiyaku Co., Ltd) JP 11029595; 1999.; Koguchi, Y.; Kohno, J.; Nishio, M.; Takahashi, K.; Okuda, T.; Ohnuki, T.; Komatsubara, S., J

Antibiot (Tokyo) 2000, 53, (2), 105-9. ; Kohno, J.; Koguchi, Y.; Nishio, M.; Nakao, K.; Kuroda, M.; Shimizu, R.; Ohnuki, T.; Komatsubara, S., J Org Chem 2000, 65, (4), 990- 5). Some macrocyclic derivatives of TMC-95 were prepared and were shown to be non covalent inhibitors of proteasome (Kaiser, M.; Groll, M.; Renner, C; Huber, R.; Moroder, L., Angew. Chem. Int. Ed. 2002, 41, (5), 780-783 ; Kaiser, M.; Siciliano, C;

Assfalg-Machleidt, L; Groll, M.; Milbradt, A. G.; Moroder, L., Org. Lett. 2003, 5, (19), 3.435-3437 ; Kaiser,M., Groll,M., Siciliano, C, Assfalg-Machleidt L, Weyher E. , Kohno J. , Milbradt A. G. , Renner G, Huber R. , Moroder L. ChemBioChem. 2004, 5, 1256-1266 ; Lin, S.; Yang, Z. Q.; Kwok, B. H.; Koldobskiy, M.; Crews, C. M.; Danishefsky, S. J., J. Am. Chem. Soc. 2004, 126, (20), 6347-6355). X-ray analysis of the complex formed between proteasome and TMC-95A proved a non-covalent binding to active sites (Groll, M.; Huber, R.; Kaiser, M.; Renner, C; Moroder, L.; Kohno, J. Crystals of proteasome-inhibitor complex. PCT Int. Appl, WO 2002081501; 2002; Groll, M.; Koguchi,Y.; Huber, R.; Kohno, J..J. MoI. Biol. 2001, 311, 543-548). Proteasome inhibitors are potential drugs by retarding or blocking the degradation of specific proteins in disorders associated with their excessive degradation. Among them, are found : inflammatory processes (Elliott et al. J Med. Chem. 2003, 81, 235- 245), various cancers (Adams J. Cancer Cell 2003, 5, 417-421 ; Adams J. Nature Reviews/Cancer 2004, 4, 349-359), immunological and auto-immune diseases (Schwartz et al. J. Immunol. 2000, 164, 6147-6157), muscle wasting (Lecker et al.

FASEBJ. 2004,18, 39-51), ischemia and cardiac pathologies (Wojcik and Napoli Stroke 2004, 35, 1506-1518), myopathies (Galbiati et al. J. Biol. Chem. 2000, 275, 37702- 37711), cystis fibrosis (Chen et al. Biochemistiy 2000, 39, 3797-3803).

Proteasome activators are susceptible to favor degradation of oxidized proteins and prevent the formation of protein aggregates as observed in Alzheimer's and

Parkinson's diseases (Cookson Ann. Neurol. 2004, 56, 315-316). Aggregated, cross- linked and oxidized proteins can inhibit 2OS proteasome (Davies Biochimie 2001, 83, 301-310). Consequently, an increase of proteasome activity can be beneficial in aging processes, noticeably in cutaneous aging. The main goal of the present invention is to provide new compounds acting as modulators of the proteasome activity (inhibitors or activators), and having the following advantages when compared to the prior art compounds mentioned above : a. they are mild, controllable and reversible inhibitors, with no creation of a covalent bond between the enzyme and the inhibitor. Due to the implication of

proteasome in a large variety of physiological processes, an irreversible permanent inhibition of proteasome would likely be detrimental. b. they are low-molecular-weight molecules and their synthetic routes are simple; c. they have a differential selectivity towards the three kinds of active sites.

The invention relates to the use of compounds of the following general formula (I) ;

wherein :

- at least one of the bonds a and b, and only one of the bonds c or d, are present, provided that :

. when the bonds a and b are present simultaneously, then R ₉ is H, and n ₅ = n ₆ = n ₇ = n ₈ = 0,

. when the bond a is present, but not the bond b, then n ₅ = n ₆ = 0, and n ₇ = n ₈ = 1, . when the bond b is present, but not the bond a, then n ₅ = n ₆ = 1 , and n ₇ = n ₈ = 0, . when the bond c is present, and d is absent, then Rg is H, . when the bond d is present, and c is absent, then Rg is an oxygen atom O,

- no is 0 or 1, and when no is 1, X = CH ₂ or X = NCH ₂ C ₆ H ₅ ,

- R ₁ is :

. OH, or a OR ₁₀ group in which Rj ₀ is a linear or branched alkyl group from 1 to 5 carbon atoms,

. or a group of formula NH-(CH ₂ )Hi-Rn in which : * u _\ = 0, or an integer from 1 to 5,

* R ₁₁ is a linear or branched alkyl group from 1 to 5 carbon atoms, an aryl group, possibly substituted, NH ₂ , or NEIR ₁₂ in which R ₁₂ is a protecting group of amine functions, such as the tertiobutyloxycarbonyl (Boc) group, or the CO-O-CH ₂ - C ₆ H ₅ (Z) group, - R ₂ is :

. H, or a linear or branched alkyl group from 1 to 5 carbon atoms, . or a group of formula (CH ₂ )H ₂ -(CO)H ₃ -NR ₁₃ R ₁₄ , in which :

* n ₂ is an integer from 1 to 5,

* n ₃ = O or 1, * R ₁₃ and R ₁₄ , independently from one another, are :

.. H,

.. or a protecting group of amine functions, such as Boc, or Z, .. or a group of formula Q=NH)NHR ₁₅ in which R ₁₅ is H or a protecting group of amine functions, such as Boc, or Z, mentioned above, . or a side chain from proteogenic aminoacids,

- R ₃ is H, or a linear or branched alkyl group from 1 to 5 carbon atoms, optionally substituted with an aryl group,

- R ₄ is H, or a protecting group of amine functions, such as Boc, or Z,

- R ₅ is H, or a protecting group of amine functions, such as Boc, or Z, - R ₆ is a OR ₁₆ group in which R ₁₆ is a linear or branched alkyl group from 1 to 5 carbon atoms,

- R ₇ and R ₈ , independently from one another, are H, or a halogen atom, such as Br, I, or Cl, as modulators of the proteasome activity, in the frame of the preparation of a medicament useful for the prevention or treatment of diseases wherein the proteasome is involved, or the preparation of cosmetic compositions, or of phytosanitary compositions.

By the expression "modulators of the proteasome activity", it must be understood that the compounds as defined above according to the present invention are: - either inhibitors of the proteasome activity, i.e. have the following inhibition properties against chymotrypsin-like, or/and trypsin-like, or/and post-acid activities of rabbit 2OS proteasome which can be measured using the appropriate fluorogenic substrate, as described below : initial rates determined in control experiments (without

test compound) were considered to be 100 % of the peptidasic activity, initial rates below 100 % were considered to be inhibitions,

- or activators of the proteasome activity, i.e. have the following activation properties against chymotrypsin-like, or/and trypsin-like, or/and post-acid activities of rabbit 2OS proteasome, which can be measured using the appropriate fluorogenic substrate as described below ; initial rates that were above 100 % in the presence of a test compound were considered to be activators.

The invention concerns more particularly the use as defined above of compounds of the following formula (II) :

in which R ₁ , R ₂ , R ₃ , and R ₄ , are such as defined above.

The invention relates more particularly to the use as defined above of compounds of formula (II) in which :

- R ₁ is a group OR ₁₀ in which Ri ₀ is a linear or branched alkyl group from 1 to 5 carbon atoms,

- R ₂ is a linear or branched alkyl group from 1 to 5 carbon atoms,

- R ₃ is a linear or branched alkyl group from 1 to 5 carbon atoms, optionally substituted with an aryl group,

- R ₄ is a protecting group of amine functions, such as Boc.

The invention also concerns more particularly the use as defined above of compounds of formula (II) in which :

- R ₁ is OCH ₃ ,

- R ₂ is CH ₃ , or CH ₂ -CH-(CH ₃ ) ₂ ,

- R ₃ is CH ₃ , or CH ₂ -C ₆ H ₅ ,

- R ₄ is Boc.

The invention relates more particularly to the use as defined above of compounds of formula (II) in which :

- R ₁ is OCH ₃ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₃ , and R ₄ is Boc (compound A374F1),

- or R ₁ is OCH ₃ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound A291),

- or R ₁ is OCH ₃ , R ₂ and R ₃ are CH ₃ , and R ₄ is Boc (compound A389Flρl2).

The invention also concerns the use as defined above of compounds of the following formula (III) :

in which R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ , are such as defined above.

The invention relates more particularly to the use as defined above of compounds of formula (III) in which:

- R ₁ is a group OR ₁₀ in which R ₁₀ is a linear or branched alkyl group from 1 to 5 carbon atoms, or a group of formula NH-(CH ₂ ) _nl -Rπ in which n _\ = O, and R ₁₁ is a linear or branched alkyl group from 1 to 5 carbon atoms,

- R ₂ is a linear or branched alkyl group from 1 to 5 carbon atoms,

- R ₃ is a linear or branched alkyl group from 1 to 5 carbon atoms, optionally substituted with an aryl group,

- R ₄ is a protecting group of amine functions, such as Boc,

- R ₅ is a protecting group of amine functions, such as Z,

- R ₆ is a OR ₁₆ group in which R ₁₆ is a linear or branched alkyl group from 1 to 5 carbon atoms.

The invention also relates more particularly to the use as defined above of compounds of formula (III) in which :

- Ri is OCH ₂ CH ₃ , OrNHCH ₃ ,

- R ₂ is CH ₃ , or CH ₂ -CH-(CH ₃ ) ₂ ,

- R ₃ is CH ₂ -C ₆ H ₅ ,

- R ₄ is un groupe Boc,

- R ₅ is un groupe Z,

- R ₆ is OCH ₃ .

The invention concerns more particularly the use as defined above of compounds of formula (III) in which :

- Ri is OCH ₂ CH ₃ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , R ₄ is Boc, R ₅ is Z, and R ₆ is OCH ₃ (compound SP221),

- or Ri is NHCH ₃ , R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , R ₄ is Boc, R ₅ is Z, and R ₆ is OCH ₃ (compound SP225F2),

- or Ri is NHCH ₃ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , R ₄ is Boc, R ₅ is Z, and R ₆ is OCH ₃ (compound SP226F1).

The invention also concerns the use as defined above of compounds of the following formula (IV):

in which c, d, n ₀ , X, R ₁ , R ₂ , R ₃ , R ₄ , R7, Rs, and R ₉ , are such as defined above.

The invention relates more particularly to the use as defined above of compounds of the following formula (W-I) :

corresponding to compounds of formula (IV) in which :

- the bond c is present, and R ₉ is H, - n ₀ = 0 or 1,

- X = CH ₂ Or NCH ₂ C ₆ H ₅ ,

- R ₁ is OH, or a group ORi ₀ in which R ₁₀ is a linear or branched alkyl group from 1 to 5 carbon atoms, or a group of formula NH-(CH ₂ ) _n i-Rπ in which m = 0, and Rn is a linear or branched alkyl group from 1 to 5 carbon atoms,

- R ₂ is H, a linear or branched alkyl group from 1 to 5 carbon atoms, or a group of formula (CH ₂ ) _n2 -(CO) _n3 -NR ₁₃ R ₁₄ , in which n ₂ = 1 to 5, n ₃ = 1, and Rj ₃ = R ₁₄ = H,

- R ₃ is a linear or branched alkyl group from 1 to 5 carbon atoms, optionally substituted with an aryl group,

- R ₄ is a protecting group of amine functions, such as Boc,

- R ₇ and Rg, independently from one another, are a halogen atom, such as Br, I. The invention concerns more particularly the use as defined above of compounds of formula (IV-I) in which :

- Ri is OH, OCH ₃ , OCH ₂ CH ₃ , Or NHCH ₃ ,

- R ₂ is H, CH ₃ , CH ₂ -CH-(CH ₃ ) ₂ ,or CH ₂ CONH ₂ ,

- R ₃ is CH ₃ , or CH ₂ -C ₆ H ₅ ,

- R ₄ is un groupe Boc,

- R ₇ is I,

- R ₈ is Br.

The invention also relates more particularly to the use as defined above of compounds of the following formula (IV-Ia) :

The invention concerns more particularly the use as defined above of compounds of formula (IV-Ia) in which :

- R ₁ is OCH ₃ , R ₂ ^' is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , R ₄ is Boc, R ₇ is I, and R ₈ is Br (compound A248), or

- R ₁ is OH, R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ Hs, R ₄ is Boc, R ₇ is I, and R ₈ is Br (compound A215), or

- R ₁ is OCH ₃ , R ₂ is CH ₂ CONH ₂ , R ₃ is CH ₂ -C ₆ H ₅ , R ₄ is Boc, R ₇ is I, and R ₈ is Br (compound SP274), or

- R ₁ is OCH ₃ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is GH ₃ , R ₄ is Boc, R ₇ is I, and R ₈ is Br (compound A363), or

- Ri is OCH ₂ CH ₃ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₃ , R ₄ is Boc, R ₇ is I, and R ₈ is Br (compound A340), or

- R ₁ is OCH ₂ CH ₃ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , R ₄ is Boc, R ₇ is I, and Rg is Br (compound Al 74), or

- R ₁ is OCH ₃ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , R ₄ is Boc, R ₇ is I, and R ₈ is Br (compound A268), or

- R ₁ is OCH ₃ , R ₂ is CH ₃ , R ₃ is CH ₃ , R ₄ is Boc, R ₇ is I, and R ₈ is Br (compound A385), or

- R ₁ is NHCH ₃ , R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , R ₄ is Boc, R ₇ is I, and R ₈ is Br (compound A254).

Preferred compounds of formula (IV-Ia) used in the frame of the present invention are compounds A215, SP274 and A254.

The invention relates more particularly to the use as defined above of compounds of the following formula (IV-Ib) :

The invention also concerns more particularly the use as defined above of compounds of formula (IV-Ib) in which :

- R ₁ is OCH ₃ , R ₂ is H, R ₃ is CH ₃ , R ₄ is Boc, R ₇ is I, R ₈ is Br, and X = CH ₂ (compound A493), or

- R ₁ is OCH ₃ , R ₂ is CH ₃ , R ₃ is CH ₃ , R ₄ is Boc, R ₇ is I, R ₈ is Br, and X = NCH ₂ C ₆ H ₅ .

The invention also relates to the use of compounds of formula (IV) as defined above wherein R ₇ and R ₈ are H.

The invention also concerns the use as defined above of compounds of the following formula (IV-2) :

corresponding to compounds of formula (IV) in which :

- the bond c is present, and R ₉ is H,

- n ₀ = 0,

- R ₁ is OH, or a group OR ₁₀ in which R _1O is a linear or branched alkyl group from 1 to 5 carbon atoms, or a group of formula NH-(CH ₂ ) _n i~Rπ in which ni = 0, or an integer from 1 to 5, and R ₁₁ is a linear or branched alkyl group from 1 to 5 carbon atoms, an aryl group, possibly substituted, NH ₂ , or NHR ₁₂ in which R ₁₂ is a protecting group of amine functions, such as Boc or Z,

- R ₂ is H, or a linear or branched alkyl group from 1 to 5 carbon atoms, or a group of formula (CH ₂ ) _n2 -(CO) _n3 -NR ₁₃ R ₁₄ , in which n ₂ is an integer from 1 to 5, n ₃ = 0 or 1, and R ₁₃ and Ri ₄ , independently from one another, are H, or a protecting group of amine functions, such as Boc, or Z, or a group of formula in which R ₁₅ is H or a protecting group of amine functions, such as Boc, or Z, mentioned above,

- R ₃ is H, or a linear or branched alkyl group from 1 to 5 carbon atoms, optionally substituted with an aryl group,

- R ₄ is a protecting group of amine functions, such as Boc, - R ₇ = R ₈ = H.

The invention concerns more particularly the use as defined above, of compounds of formula (IV-2) in which :

- n ₀ = 0,

- R ₁ is OH, OCH ₃ , NHCH ₂ C ₆ H ₅ , NHC ₆ H ₅ , NHC ₆ H ₄ OH, or NH(CH ₂ ) ₄ NHBoc,

- R ₂ is H, CH ₃ , CH ₂ -CH-(CHs) ₂ , CH ₂ CONH ₂ , (CH ₂ ) ₃ NHC(=NH)NH ₂ , or (CH ₂ ) ₃ NZC(=NH)NHZ, or (CH ₂ ) ₄ NHBoc,

- R ₃ is H ₅ or CH ₂ -C ₆ H ₅ ,

- R ₄ is Boc.

The invention relates more particularly to the use as defined above of compounds of formula (IV-2) in which : - R ₁ is NHCH ₂ C ₆ H ₅ , R ₂ is H, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound

PSVI lR),

- or R ₁ is OH, R ₂ is H, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound NR35),

- or R ₁ is NHC ₆ H ₅ , R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP303r2), - or R ₁ is NHCH ₂ C ₆ H ₅ , R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound

SP304R),

- or R ₁ is NHC ₆ H ₄ OH, R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP313P),

- or R ₁ is NH(CH ₂ ) ₄ NHBoc, R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP305R),

- or R ₁ is OCH ₃ , R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound NR36),

- or R ₁ is OCH ₃ , R ₂ is CH ₃ , R ₃ is H, and R ₄ is Boc (compound NR40),

- or R ₁ is NHC ₆ H ₅ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound A424P), - or R ₁ is NHCH ₂ C ₆ H ₅ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc

(compound A414P),

- or R ₁ is NHC ₆ H ₄ OH, R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound A418P),

- or R ₁ is NH(CH ₂ ) ₄ NHBoc, R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP296P),

- or R ₁ is NHC ₆ H ₅ , R ₂ is CH ₂ CONH ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP314C2),

- or R ₁ is NHCH ₂ C ₆ H ₅ , R ₂ is CH ₂ CONH ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound A416), - or R ₁ is NHC ₆ H ₄ OH, R ₂ is CH ₂ CONH ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc

(compound SP318C),

- or R ₁ is NH(CH ₂ ) ₄ NHBoc, R ₂ is CH ₂ CONH ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP323C2),

- or R ₁ is NHC ₆ H ₅ , R ₂ is (CH ₂ ) ₃ NHC(=NH)NH ₂ , R ₃ is H, and R ₄ is Boc (compound SP325),

- or R ₁ is NHC ₆ H ₄ OH, R ₂ is (CH ₂ ) ₃ NHC(=NH)NH ₂ , R ₃ is H, and R ₄ is Boc (compound SP324), - or R ₁ is NHC ₆ H ₅ , R ₂ is (CH ₂ ) ₃ NZC(=NH)NHZ, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc

(compound SP31 OC),

- or R ₁ is NHCH ₂ C ₆ H ₅ , R ₂ is (CH ₂ ) ₃ NZC(=NH)NHZ, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP315C2),

- or R ₁ is NHC ₆ H ₄ OH, R ₂ is (CH ₂ ) ₃ NZC(=NH)NHZ, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP320P2),

- or R ₁ is NH(CH ₂ ) ₄ NHBoc, R ₂ is (CH ₂ ) ₃ NZC(=NH)NHZ, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP311C),

- or R ₁ is NHC ₆ H ₅ , R ₂ is (CH ₂ ) ₄ NHBoc, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP306P), - or R ₁ is NHCH ₂ C ₆ H ₅ , R ₂ is (CH ₂ ) ₄ NHBoc, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc

(compound SP307P),

- or R ₁ is NHC ₆ H ₄ OH, R ₂ is (CH ₂ ) ₄ NHBoc, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP319P),

- or R ₁ is NH(CH ₂ ) ₄ NHBoc, R ₂ is (CH ₂ ) ₄ NHBoc, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP308P).

- or R ₁ is OH, R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound NR66).

- or R ₁ is OH, R ₂ is CH ₃ , R ₃ is H, and R ₄ is Boc (compound NR68).

Preferred compounds of formula (IV-2) used in the frame of the present invention are the compounds SP313P, NR40, SP325, and SP324. The invention also concerns the use as defined of compounds of the following formula (IV-3) :

corresponding to compounds of formula (IV) in which :

- the bond d is present, and Rg is an oxygen atom O,

- n ₀ = 0,

- R ₁ is OH, or a group OR ₁₀ in which R ₁₀ is a linear or branched alkyl group from 1 to 5 carbon atoms, or a group of formula NH-(CH2) _nl -Rn in which H ₁ = 0, or an integer from 1 to 5, and R ₁₁ is an aryl group, possibly substituted,

- R ₂ is H, or a linear or branched alkyl group from 1 to 5 carbon atoms, or a group of formula (CH ₂ ) _n2 -(CO) _n3 -NRi ₃ R ₁₄ , in which n ₂ is an integer from 1 to 5, n ₃ = 0 or 1, and R ₁₃ and R ₁₄ , independently from one another, are H, or a protecting group of amine functions, such as Boc, or Z, or a group of formula C(^NH)NHR ₁₅ in which Rj ₅ is H or a protecting group of amine functions, such as Boc, or Z, mentioned above,

- R ₃ is H, or a linear or branched alkyl group from 1 to 5 carbon atoms, optionally substituted with an aryl group,

- R ₄ is a protecting group of amine functions, such as Boc, - R ₇ = R ₈ = H.

The invention relates more particularly to the use as defined above of compounds of formula (IV-3) in which :

- R ₁ is OH, OCH ₃ , NHCH ₂ C ₆ H ₅ , or NHC ₆ H ₅ ,

- R ₂ is H, CH ₃ , CH ₂ -CH-(CH ₃ ) ₂ , (CH ₂ ) ₃ NZC(=NH)NHZ, or (CH ₂ ) ₄ NHBoc,

- R ₃ is H, or CH ₂ -C ₆ H ₅ ,

- R ₄ is Boc.

The invention also relates more particularly to the use as defined above of compounds of formula (IV-3) in which :

- R ₁ is NHC ₆ H ₅ , R ₂ is (CH ₂ ) ₃ NZC(=NH)NHZ, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound CVl 1), - R ₁ is NHC ₆ H ₅ , R ₂ is (CH ₂ ) ₄ NHBoc, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound

CV12),

- R ₁ is NHC ₆ H ₅ , R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound CV13),

- Ri is NHC ₆ H ₅ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound JV602), - Ri is NHCH ₂ C ₆ H ₅ , R ₂ is H, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound NRl 5),

- Ri is OCH ₃ , R ₂ is H, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound NR38),

- Ri is NHCH ₂ C ₆ H ₅ , R ₂ is (CH ₂ ) ₃ NZC(=NH)NHZ, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound NRl 6),

- R ₁ is OCH ₃ , R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc, - Ri is OH, R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc,

- Ri is OH, R ₂ is CH ₃ , R ₃ is H, and R ₄ is Boc,

- R ₁ is OCH ₃ , R ₂ is CH ₃ , R ₃ is H, and R ₄ is Boc.

Preferred compounds of formula (IV-3) used in the frame of the present invention are the compounds CV12, CV13, NR15, and NR38. The invention relates more particularly to the use of a compound as defined above, as modulators of the proteasome activity for the preparation of:

- a drug for prevention or treatment of pathologies involving proteasome, said pathologies being chosen from the group constituted by: cancers involving haematological or solid tumors, immunological diseases, auto-immune diseases, AIDS, inflammatory diseases, cardiac pathologies and consequences of ischemic processes in myocardial, cerebral or pulmonary regions, allograft rejection, myopathies, muscle wasting, cerebrovascular accidents, traumatisms, burns, pathologies associated with aging like Alzheimer's disease and Parkinson's disease, and the appearance of aging signs, or - a drug for increasing the radiosensitization of a tumor, the sensitivity to chemotherapy and/or immunotherapy, or promoting the circumvention of resistances, or

- a cosmetic composition for the implementation of a method of cosmetic prevention or treatment of the appearance of cutaneous aging and/or photoaging, or

o

- phytosanitary compositions for the implementation of processes for modulating the defence response of plants, in particular phytosanitary compositions for the stimulation of plants defence response against phytopathogenic agents.

Advantageously pharmaceutical' compositions or drags used in the frame of the present invention comprise compounds as defined above mainly acting as inhibitors of the proteasome activity.

Preferred compounds contained in the pharmaceutical compositions as defined above are those of formula IV-IA, or TV-2, or IV-3, or IV-IB such as compounds A215, SP274, A254, or SP313P, NR40, SP325, SP324, or CV12, CV13, NR15, NR38, or A493.

Advantageously cosmetic compositions used in the frame of the present invention comprise compounds as defined above mainly acting as activators of the proteasome activity.

Preferred compounds contained in the cosmetic compositions as defined above are those of formula II, or III, or IV-IA, or IV-2, or IV-3, or TV-IB such as compounds

A374F1, or SP221, or A363, or NR36, SP305R, SP314C2, or NRl 5, NR38, NR16, or A493.

Advantageously phytosanitary compositions used in the frame of the present invention comprise compounds as defined above mainly acting as inhibitors of the proteasome activity.

Preferred compounds contained in the phytosanitary compositions as defined above are those of formula formula IV-IA, or IV-2, or IV-3, or IV-IB such ^' as compounds A215, SP274, A254, or SP313P, NR40, SP325, SP324, or CV12, CV13, NR15, NR38, or A493. The invention also concerns a pharmaceutical composition, characterized in that it comprises a compound as defined above, in association with a pharmaceutically acceptable vehicle.

The invention relates more particularly to the pharmaceutical composition as defined above, characterized in that it contains a compound as defined above, at an appropriate amount for a daily administration of about twice a week for 4 weeks at about 1.5 mg/m ² .

The invention also relates more particularly to the pharmaceutical composition as defined above, characterized in that it is in a form suitable for intravenous or per os administration.

The invention also concerns a cosmetic composition characterized in that it comprises a compound as defined above, in association with a pharmacologically acceptable vehicle.

The invention relates more particularly to the cosmetic composition as defined above, characterized in that it is in a form suitable for dermatological administration, in particular as a cream, pomade or gel.

The invention concerns more particularly the cosmetic composition as defined above, characterized in that it contains a compound as defined above, at an appropriate amount for a daily administration of about 1 mg/m to 10 mg/m of skin.

The invention also concerns a phytosanitary composition, characterized in that it comprises a compound as defined above, if necessary in association with an acceptable vehicle in phytosanitary field.

The invention relates more particularly to the phytosanitary composition as defined above, characterized in that it comprises a compound as defined above, at an appropriate amount for an administration by spraying of about 1 g/m ² to 10 g/m ² .

The invention also concerns the compounds of formula (I), and more particularly of formula (II), (III), and (IV) as defined above.

The invention also relates to compounds of the following formula (III) :

in which R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ , are such as defined above, the compound of formula (III) in which R ₁ = NB(CH ₂ ) ₂ CH ₃ , R ₂ = CH ₂ CONH ₂ , R ₃ = CH ₃ , R ₄ = Z, R ₅ = Boc, R ₆ = OtBu being excluded.

The invention relates more particularly to compounds of formula (III) as defined above in which :

- R ₁ is a group OR ₁₀ in which R ₁₀ is a linear or branched alkyl group from 1 to 5 carbon atoms, or a group of formula NH-(CHa) _n I-Rn in which m = 0, and R ₁₁ is a linear or branched alkyl group from 1 to 5 carbon atoms,

- R ₂ is a linear or branched alkyl group from 1 to 5 carbon atoms, - R ₃ is a linear or branched alkyl group from 1 to 5 carbon atoms, optionally substituted with an aryl group,

- R ₄ is a protecting group of amine functions, such as Boc,

- R ₅ is a protecting group of amine functions, such as Z,

- R ₆ is a OR ₁₆ group in which R ₁₆ is a linear or branched alkyl group from 1 to 5 carbon atoms.

The invention also relates more particularly to compounds of formula (III) as defined above in which :

- R ₁ is OCH ₂ CH ₃ , OrNHCH ₃ ,

- R ₂ is CH ₃ , or CH ₂ -CH-(CH ₃ ) ₂ , - R ₃ Is CH ₂ -C ₆ H ₅ ,

- R ₄ is Boc,

- R ₅ is Z, - R ₆ is OCH ₃ .

The invention concerns more particularly the compounds of formula (III) as defined above in which :

- R ₁ is OCH ₂ CH ₃ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , R ₄ is Boc, R ₅ is Z, and R ₆ is OCH ₃ (compound SP221),

- or R ₁ is NHCH ₃ , R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , R ₄ is Boc, R ₅ is Z, and R ₆ is OCH ₃ (compound SP225F2), - or R ₁ is NHCH ₃ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , R ₄ is Boc, R ₅ is Z, and

R ₆ is OCH ₃ (compound SP226F1).

The invention also relates to compounds as defined above, of the following formula (IV) :

in which c, d, no, X, R ₁ , R ₂ , R ₃ , R ₄ , R ₇ , R ₈ , and Rg, are such as defined above, the compounds of formula (IV) in which n ₀ , = O, R ₉ = H, R ₁ = OCH ₃ , R ₄ is Boc, R ₇ is I, R ₈ is Br, and

- R ₂ is CH ₃ , and R ₃ is CH ₂ -C ₆ H ₅ (compound A248), or

- R ₂ is CH ₂ CONH ₂ , and R ₃ is CH ₂ -C ₆ H ₅ (compound SP274), or

- R ₂ is CH ₂ -CH-(CH ₃ );., and R ₃ is CH ₃ (compound A363), or

- R ₂ is CH ₂ -CH-(CHs) ₂ , and R ₃ is CH ₂ -C ₆ H ₅ (compound A268), being exluded.

The invention relates more particularly to compounds as defined above, of the following formula (IV-I) :

corresponding to compounds of formula (IV) in which : - the bond c is present, and R ₉ is H,

- n ₀ = 0 or 1,

- X = CH ₂ Or NCH ₂ C ₆ H ₅ ,

- R ₁ is OH, or a group OR ₁₀ in which R ₁₀ is a linear or branched alkyl group from 1 to 5 carbon atoms, or a group of formula NH-(CH ₂ )Hi-R ₁ 1 in which m = 0, and Rn is a linear or branched alkyl group from 1 to 5 carbon atoms,

- R ₂ is H, a linear or branched alkyl group from 1 to 5 carbon atoms, or a group of formula (CH ₂ ) _n2 -(CO) _n3 -NR ₁₃ R ₁₄ , in which n ₂ = 1 to 5, n ₃ = 1, and R ₁₃ = Ri ₄ = H,

- R ₃ is a linear or branched alkyl group from 1 to 5 carbon atoms, optionally substituted with an aryl group,

- R ₄ is a protecting group of amine functions, such as Boc,

- R ₇ and R ₈ , independently from one another, are a halogen atom, such as Br, I. The invention concerns more particularly compounds of formula (IV-I) as defined above in which :

- R ₁ is OH, OCH ₃ , OCH ₂ CH ₃ , OrNHCH ₃ ,

- R ₂ is H, CH ₃ , CH ₂ -CH-(CH ₃ ) ₂ ,or CH ₂ CONH ₂ ,

- R ₃ is CH ₃ , or CH ₂ -C ₆ H ₅ ,

- R ₄ is Boc, - R ₇ is I,

- R ₈ is Br.

The invention relates more particularly to compounds as defined above, of the following formula (IV-Ia):

The invention also relates more particularly to compounds of formula (IV-Ia) as defined above in which :

- I _-0 = O, R ₁ is OH, R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , R ₄ is Boc, R ₇ is I, and R ₈ is Br (compound A215),

- or n ₀ = 0, R ₁ is OCH ₂ CH ₃ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₃ , R ₄ is Boc, R ₇ is I, and R ₈ is Br (compound A340),

- or n ₀ = 0, R ₁ is OCH ₂ CH ₃ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , R ₄ is Boc, R ₇ is I, and R ₈ is Br (compound Al 74),

- or n ₀ = 0, Ri is OCH ₃ , R ₂ is CH ₃ , R ₃ is CH ₃ , R ₄ is Boc, R ₇ is I, and R ₈ is Br (compound A385),

- or n ₀ = 0, R ₁ is NHCH ₃ , R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , R ₄ is Boc, R ₇ is I, and R ₈ is Br (compound A254).

The invention relates more particularly to compounds as defined above of the following formula (W-Ib):

The invention concerns more particularly compounds of formula (IV-Ib) as defined a bove in which

- R ₁ is OCH ₃ , R ₂ is H, R ₃ is CH ₃ , R ₄ is Boc, R ₇ is I, R ₈ is Br, and X = CH ₂ (compound A493), or

- R ₁ is OCH ₃ , R ₂ is CH ₃ , R ₃ is CH ₃ , R ₄ is Boc, R ₇ is I, R ₈ is Br, and X = NCH ₂ C ₆ H ₅ .

The invention also relates to compounds of formula (IV) as defined above wherein R ₇ and R ₈ are H.

The invention also concerns compounds as defined above of the following formula (IV-2) :

corresponding to compounds of formula (IV) in which :

- the bond c is present, and Rg is H, - Ho = O,

- R ₁ is OH, or a group OR _1O in which R ₁₀ is a linear or branched alkyl group from 1 to 5 carbon atoms, or a group of formula NH-(CH ₂ ) _nl -Rπ in which n\ = O ₃ or an integer from 1 to 5, and R ₁₁ is a linear or branched alkyl group from 1 to 5 carbon atoms, an aryl group, possibly substituted, NH ₂ , or NHR ₁₂ in which R ₁₂ is a protecting group of amine functions, such as Boc or Z,

- R ₂ is H, or a linear or branched alkyl group from 1 to 5 carbon atoms, or a group of formula (CH ₂ ) _n2 -(CO) _n3 -NR ₁₃ R ₁₄ , in which n ₂ is an integer from 1 to 5, n ₃ = 0 or 1, and R ₁₃ and R ₁₄ , independently from one another, are H, or a protecting group of amine functions, such as Boc, or Z, or a group of formula C(^NH)NHR ₁₅ in which R ₁₅ is H or a protecting group of amine functions, such as Boc, or Z, mentioned above,

- R ₃ is H, or a linear or branched alkyl group from 1 to 5 carbon atoms, optionally substituted with an aryl group,

- R ₄ is a protecting group of amine functions, such as Boc,

- R ₇ = R ₈ = H.

The invention relates more particularly to compounds of formula (IV-2) as defined above in which :

- R ₁ is OH, OCH ₃ , NHCH ₂ C ₆ H ₅ , NHC ₆ H ₅ , NHC ₆ H ₄ OH ₅ Or NH(CH ₂ ) ₄ NHBoc,

- R ₂ is H, CH ₃ , CH ₂ -CH-(CH ₃ ) ₂ , CH ₂ CONH ₂ , (CH ₂ ) ₃ NHC(=NH)NH ₂ , or (CH ₂ ) ₃ NZC(=NH)NHZ, or (CH ₂ ) ₄ NHBoc,

- R ₃ is H, or CH ₂ -C ₆ H ₅ ,

- R ₄ is Boc. The invention concerns more particularly compounds of formula (IV-2) as defined above in which :

- Ri is NHCH ₂ C ₆ H ₅ , R ₂ is H, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound PSVIlR),

- or Ri is OH, R ₂ is H, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound NR35), - or R ₁ is NHC ₆ H ₅ , R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound

SP303r2),

- or R ₁ is NHCH ₂ C ₆ H5, R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP304R),

- or R ₁ is NHC ₆ H ₄ OH, R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP313P),

- or R ₁ is NH(CH ₂ ) ₄ NHBoc, R ₂ is CH ₃ , R ₃ is CH ₂ -C ₅ H ₅ , and R ₄ is Boc (compound SP305R),

- or R ₁ is OCH ₃ , R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound NR36),

- or R ₁ is OCH ₃ , R ₂ is CH ₃ , R ₃ is H, and R ₄ is Boc (compound NR40), - or R ₁ is NHC ₆ H ₅ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc

(compound A424P),

- or Ri is NHCH ₂ C ₆ H ₅ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound A414P),

- or Ri is NHC ₆ H ₄ OH, R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound A418P),

- or R ₁ is NH(CH ₂ ) ₄ NHBoc, R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP296P),

- or R ₁ is NHC ₆ H ₅ , R ₂ is CH ₂ CONH ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP314C2), - or Ri is NHCH ₂ C ₆ H ₅ , R ₂ is CH ₂ CONH ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc

(compound A416),

- or R ₁ is NHC ₆ H ₄ OH, R ₂ is CH ₂ CONH ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP318C),

- or R ₁ is NH(CH ₂ ) ₄ NHBoc, R ₂ is CH ₂ CONH ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP323C2),

- or R ₁ is NHC ₆ H ₅ , R ₂ is (CH ₂ ) ₃ NHC(=NH)NH ₂ , R ₃ is H, and R ₄ is Boc (compound SP325), - or R ₁ is NHC ₆ H ₄ OH, R ₂ is (CH ₂ ) ₃ NHC(=NH)NH ₂ , R ₃ is H, and R ₄ is Boc

(compound SP324),

- or R ₁ is NHC ₆ H ₅ , R ₂ is (CH ₂ ) ₃ NZC(=NH)NHZ, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP31 OC),

- or R ₁ is NHCH ₂ C ₆ H ₅ , R ₂ is (CH ₂ ) ₃ NZC(=NH)NHZ, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP315C2),

- or Ri is NHC ₆ H ₄ OH, R ₂ is (CH ₂ ) ₃ NZC(=NH)NHZ, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP320P2),

- or R ₁ is NH(CH ₂ ) ₄ NHBoc, R ₂ is (CH ₂ ) ₃ NZC(=NH)NHZ, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP311C), - or R ₁ is NHC ₆ H ₅ , R ₂ is (CH ₂ ) ₄ NHBoc, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc

(compound SP306P),

- or R ₁ is NHCH ₂ C ₆ H ₅ , R ₂ is (CH ₂ ) ₄ NHBoc, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP307P),

- or R ₁ is NHC ₆ H ₄ OH, R ₂ is (CH ₂ ) ₄ NHBoc, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP319P),

- or R ₁ is NH(CH ₂ ) ₄ NHBoc, R ₂ is (CH ₂ ) ₄ NHBoc, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound SP308P).

- or R ₁ is OH, R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound NR66).

- or R ₁ is OH, R ₂ is CH ₃ , R ₃ is H, and R ₄ is Boc (compound NR68). The invention relates more particularly to compounds SP313P, NR40, SP325, and

SP324, as preferred compounds of formula (IV -2).

The invention also concerns compounds as defined above, of the following formula (IV-3) :

corresponding to compounds of formula (IV) in which :

- the bond d is present, and Rg is an oxygen atom O,

- n ₀ = 0,

- R ₁ is OH, or a group OR ₁ O in which R ₁ O is a linear or branched alkyl group from 1 to 5 carbon atoms, or a group of formula NH-(CH ₂ ) _nl -Rπ in which Ά _\ - 0, or an integer from 1 to 5, and R ₁₁ is an aryl group, possibly substituted,

- R ₂ is H, or a linear or branched alkyl group from 1 to 5 carbon atoms, or a group of formula (CH ₂ ) _H2 -(CO) ₁₁₃ -NR ₁₃ R ₁₄ , in which n ₂ is an integer from 1 to 5, n ₃ = 0 or 1, and R ₁₃ and R ₁₄ , independently from one another, are H, or a protecting group of amine functions, such as Boc, or Z, or a group of formula C^NH)NHR ₁₅ in which R ₁₅ is H or a protecting group of amine functions, such as Boc, or Z, mentioned above,

- R ₃ is H, or a linear or branched alkyl group from 1 to 5 carbon atoms, optionally substituted with an aryl group,

- R ₄ is a protecting group of amine functions, such as Boc, - R ₇ = R ₈ = H.

The invention relates more particularly to compounds of formula (IV-3) as defined above in which :

- R ₁ is OH, OCH ₃ , NHCH ₂ C ₆ H ₅ , OrNHC ₆ H ₅ ,

- R ₂ is H, CH ₃ , CH ₂ -CH-(CHs) ₂ , (CH ₂ ) ₃ NZC(=NH)NHZ, or (CH ₂ ) ₄ NHBoc,

- R ₃ is H, or CH ₂ -C ₆ H ₅ ,

- R ₄ is Boc.

The invention also relates more particularly to compounds of formula (IV-3) as defined above in which :

- R ₁ is NHC ₆ H ₅ , R ₂ is (CH ₂ ) ₃ NZC(=NH)NHZ, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound CVIl), - R ₁ is NHC ₆ H ₅ , R ₂ is (CH ₂ ) ₄ NHBoc, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound

CV12),

- R ₁ is NHC ₆ H ₅ , R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound CV13),

- R ₁ is NHC ₆ H ₅ , R ₂ is CH ₂ -CH-(CH ₃ ) ₂ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound JV602), - R ₁ is NHCH ₂ C ₆ H ₅ , R ₂ is H, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound NRl 5),

- R ₁ is OCH ₃ , R ₂ is H, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound NR38),

- R ₁ is NHCH ₂ C ₆ H ₅ , R ₂ is (CH ₂ ) ₃ NZC(==NH)NHZ, R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc (compound NRl 6),

- R ₁ is OCH ₃ , R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc, - R ₁ is OH, R ₂ is CH ₃ , R ₃ is CH ₂ -C ₆ H ₅ , and R ₄ is Boc,

- Ri is OH, R ₂ is CH ₃ , R ₃ is H, and R ₄ is Boc,

- R ₁ is OCH ₃ , R ₂ is CH ₃ , R ₃ is H, and R ₄ is Boc.

The invention relates more particularly to compounds CV12, CV13, NR15, and NR38, as preferred compounds of formula (IV-3). Advantageously compounds of formula (II), (III), (IV-I), (IV-2), and (IV-3) as defined above are obtained according to the following retrosynthetic scheme :

)

)

Synthons (V), (VI) and (VII) were assembled according to standard peptide synthesis (Bodansky, M.; Bodansky, A., The practice of peptide synthesis. Springer Verlag, 1995) or according to techniques of biaryl synthesis (Hassan, J.; Sevignon, M.; Gozzi, C; Schulz, E.; Lemaire, M., Chem. Rev. 2002, 102, (5), 1359-1469). The following reaction pathways were used:

(vπi) ( V ) biaryl synthesis

( III )

R ₇ = I R ₈ = Br

R ₉ = H c present

(Vffl ) ( V ) peptidic synthesis (IV)

(IV-I) biaryl synthesis _m ~~

The invention will be further illustrated with the detailed description wich follows of the synthesis and the biological properties of compounds of the invention.

I) Preparation of starting material (synthons V and VIII)

A) Preparation of 7-bromotryptophane derivatives

HCl ₅ (7-bromo)Trp-OCH ₃ :

prepared according to Berthelot, A.; Piguel, S.; Le Dour, G.; Vidal, J., Synthesis of macrocyclic peptide analogues of proteasome inhibitor TMC-95A. J. Org. Chem. 2003, 68, (25), 9835-9838.

HCl, (7-bromo)Trp-OEt:

(7-bromo)Trp(Boc)-OtBu (55 mg, 0.13 mmol, prepared according to Berthelot, A.; Piguel, S.; Le Dour, G.; Vidal, J., Synthesis of macrocyclic peptide analogues of proteasome inhibitor TMC-95A. J. Org. Chem. 2003, 68, (25), 9835-9838) was dissolved in a 3 M solution of anhydrous HCl in EtOH (0.5 mL). Evaporation of the solvent to dryness gave the crude HCl, (7-bromo)Trp-OEt (45 mg, 100%) as a white solid which was used without further purification. ¹ H NMR (200 MHz, D ₂ O) δ 1.05 (t, J = 7.1 Hz, 3H, CH ₃ ), 3;36 (d, J - 6.4 Hz, 2H, CH ₂ ), 4.21 (q, J = 7.1 Hz, 2H, CH ₂ (Et)), 4.31 (t, J = 6.1 Hz, IH, CH), 6.99 (t, J = 7.7 Hz, IH, H5), 7.26 (s, IH, H2), 7.36 (d, J = 7.7 Hz, IH, H6) and 7.49 (d, J = 7.7 Hz, IH, H4). ¹³ C NMR (75 MHz, CD ₃ OD) δ 14.3 (CH ₃ ), 27.6 (CH ₂ (Et)), 54.7 (CHa), 63.7 (CH ₂ ), 105.9 (C(7)), 109.1 (C(3)), 118.5 (CH), 121.6 (CH), 125.5 (CH), 126.9 (CH), 129.9 (C), 136.6 (C) and 170.3 (CO).

Z-(7-bromo)Trp-OH:

To a solution of (7-bromo)Trp(Boc)-OtBu (347 mg, 0.79 mmol, prepared according to Berthelot, A.; Piguel, S.; Le Dour, G.; Vidal, J., Synthesis of macrocyclic peptide analogues of proteasome inhibitor TMC-95A. J. Org. Chem. 2003, 68, (25), 9835-9838.), in DMF (2 mL) cooled at 0 °C was added ZOSu (217 mg, 0.87 mmol). The resulting mixture was stirred 1 h at O ⁰ C and 1 h 30 at room temperature before concentration of the solvents. The residue was then diluted in CH ₂ Cl ₂ and washed with water. The organic layer was then dried over Na ₂ SO ₄ and the solvent was removed in vacuo. Purification by column chromatography (10% AcOEt/heptane) gave Z-(7-bromo)Trρ(Boc)-OtBu as a light yellow oil (435 mg, 96 %). Rf 0.44 (20% AcOEt/heptane). ¹ H NMR (300 MHz, CD ₃ Cl ₃ ) δ 1.39 (s, 9H, tBu), 1.65 (s, 9H, 3CH ₃ (BoC)), 3.2 (d, J = 5.5 Hz, 2H, CH ₂ ), 4.61-4.64 (m, IH, CH), 5.13 (AB, J = 12.2 Hz, 2H, CH ₂ (Z)), 5.36 ( broad d, J = 7.8 Hz, IH, NH(Z)), 7.06 (t, J = 7.8 Hz, IH, H5), 7.32-7.37 (m, 6H, 5 aromatic H (Z) and H2) and 7.49-7.55 (m, 2H, H4 and H6). Then, Z-(7- bromo)Trp(Boc)-OtBu (775 mg, 1.35 mmol) was dissolved in a 3 M solution of anhydrous HCl in AcOEt (3 mL). Evaporation of the solvent to dryness gave the crude Z-(7-bromo)Trp-

OH (564 mg, 100%) which was used without further purification. ¹ H NMR (200 MHz, CD ₃ OD) δ 3.14 (dd, J = 14.4 Hz, J = 8 Hz, IH) and 3;32 (dd, J = 14.4 Hz, J = 5.6 Hz, IH) CH ₂ , 4.54 (dd, J = 8 Hz, J = 5.6 Hz, IH, CH), 5.05 (d, J = 2.6 Hz, 2H, CH ₂ (Z)), 6.94 (t, J = 7.8 Hz, IH, H5), 7.17 (s, IH, H2), 7.27-7.39 (m, 6H, 5 aromatic H(Z) and H6) and 7.53 (d, J = 7.8 Hz, IH, H4). ¹³ C NMR (50 MHz ₅ CD ₃ OD) δ 29.2 (CH ₂ ), 56.6 (CHa), 57.9 (CH ₂ Z), 105.9 (C), 112.9 (C), 119.2 (CH), 121.5 (CH), 125.4 (CH), 126.1 (CH), 129.1 (CH(Z)), 129.3 (CH(Z)), 129.8 (CH(Z)), 130.8 (C(Z)), 136.8 (C), 138.5 (C), 158.7 (C(Z)) and 175.8 (C).

Z-(7-bromo)Trp-OEt:

Z-7-bromo-Trp(Boc)-OtBu (2.69 g, 4.69 g, obtained as described for Z-(7-bromo)Trp-OH) was dissolved in a HCl/AcOEt/EtOH mixture and allowed to react for 21 h. Evaporation of the solvent to dryness gave the crude Z-(7-bromo)Trp-OEt which was purified by flash chromatography on silica gel (3% MeOH/CH ₂ Cl ₂ ) (1.44 g, 70%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 1.21 (t, J = 7.1 Hz, 3H, CH ₃ ), 3.31 (m, 2H, CH ₂ Trp), 4.11 (m, 2H, CH ₂ OEt), 4.71 (m, IH, CHa), 5.09 and 5.16 (AB system, J = 12.2 Hz, 2H), 5.33 (d, J = 8.0 Hz, IH, NHZ), 6.98 (t, J = 7.8 Hz, IH), 7.05 (d, J = 2.0 Hz, IH), 7.34 (m, 6H), 7.49 (d, J = 7.8 Hz ₃ IH), 8.20 (s, IH, indolic NH). ¹³ C NMR (75 MHz, CDCl ₃ ) δ 14.5 (CH ₃ ), 28.6 (CH ₂ Trp), 54.9 (CHa), 62.0 (CH ₂ ), 67.3 (CH ₂ ), 105.2 (C), 111.8 (C), 118.4 (CH), 121.2 (CH), 123.8 (CH), 124.9 (CH), 128.5 (CH), 128.6 (CH), 128.9 (CH), 129.2 (C), 135.2 (C), 136.7 (C), 156.1 (CO Z), 172.1 (CO ester). Anal. Calcd. for C ₂₁ H ₂₁ BrN ₂ O ₄ : C, 56.64; H, 4.75; N, 6.29. Found: C, 57.29; H, 5.03; N, 5.93.

Z-(7-bromo)Trp-NHMe:

To a solution of Z-(7-bromo)Trp-OH (784 mg, 1.88 mmol) in THF (10 mL) cooled at 0 °C was added dropwise NEt ₃ (315 μL, 2.26 mmol) and ethyl chloroformiate (215 μL, 2.26mmol). The resulting solution was stirred 20 mn before methylamine 2M solution in THF (3 mL, 6 mmol) was added. The mixture was stirred a further 2 h 30 and evaporated to dryness. The residue was suspended in water before being collected by filtration. Z-(7-

bromo)Trp-NHMe was obtained as a white solid (683 mg, 85%). mp(dec) = 200°C. ¹ H NMR (200 MHz, CD ₃ OD) δ 2.61 (d, J = 4.5 Hz, 3H, CH ₃ ), 2.91 (dd, J = 15.1 Hz ₃ J = 4 Hz, IH) and 3;10 (dd, J = 15.1 Hz, J = 10.1 Hz, IH) CH ₂ , 4.14-4.31 (m, IH, CH), 4.96 (s, 2H, CH ₂ (Z)), 6.96 (t, J = 7.7 Hz, IH, H5), 7.23-7.41 (m, 6H, 5 aromatic H(Z) and H2), 7.46 (d, J = 7.7 Hz, IH ₅ H6), 7.67 (d, J - 7.7 Hz, IH, H4), 8.0 (broad s, IH, NH(Me)) and 11.09 (broad s, IH, NH). ¹³ C NMR (50 MHz, DMSO-d ⁶ ) δ 26.0 (CH ₃ ), 28.4 (CH ₂ ), 56.5 (CHa), 65.6 (CH ₂ Z), 104.5 (C), 112.2 (C), 118.5 (CH), 120.1 (CH), 123.8 (CH), 125.6 (CH), 127.8 (CH(Z)), 128.0 (CH(Z)), 128.6 (CH(Z)), 129.4 (C(Z)), 134.7 (C), 137.4 (C), 156.2 (C(Z)) and 172.4 (C). HRMS (FAB) calcd for C ₂₀ H ₂₀ BrN ₃ O ₃ [M+H ⁺ ] 430.0766, found 430.0766.

HBr, (7-bromo)Trp-NHMe:

Z-(7-bromo)Trp-NHMe (376 mg, 0.87 mmol) was dissolved in an HBr 45% w/v solution in acetic acid (1 rnL) and was stirred at room temperature for 8 h and evaporated to dryness. The residue, solubilized in water, was lyophilized and gave HBr, (7-bromo)Trp-NHMe (330 mg, 100%) as a brown solid which was used without purification. ¹ H NMR (200 MHz, DMSO- d ⁶ ) δ 2.64 (d, 2H, J = 5 Hz, CH ₃ ), 3.12-3.21 (m, 2H, CH ₂ ), 3.92 (m, IH, CH), 7.0 (t, J = 7.7 Hz, IH, H5), 7.29 (d, J = 2.4 Hz, IH, H2), 7.35 (d, J = 7.7 Hz, IH, H6), 7.67 (d, J = 7.7 Hz, IH, H4), 8.09 (broad s, 3H ₅ NH ₃ ⁺ ), 8.48 (broad d, J = 5 Hz ₅ IH, NH(Me)) and 11.28 (broad s, IH, NH).

B) Preparation of tryptophane derivatives

Z-Trp-NHPh: To a solution of Z-Trp-OH (2 g, 5.9 mmol) in THF (88 mL) at 5 ⁰ C were added aniline (540 μL, 5.9 mmol) and DCC (1.6 g, 7.7 mmol). The reaction was allowed to warm up to room temperature overnight. The solvent was evaporated off and the crude was triturated with ethyl acetate (50 mL). After filtration, the organic phase was successively washed with aqueous 5% KHSO ₄ , aqueous 10% KHCO ₃ , brine and was dried over Na ₂ SO ₄ .

The solvent was removed in vacuo. The crude amide was purified by precipitation in methanol/pentane to give a white amorphous solid (1.7 g, 69%). ¹ H NMR (300 MHz, CDCl ₃ )

δ 3.25 (dd, J = 14.4 Hz, J = 8 Hz, IH, CH ₂ ), 3.46 (dd, J = 14.4 Hz, J = 5.3 Hz, IH, CH ₂ ), 4.67 (m, IH, CHa), 5.12 (m, 2H, CH ₂ (Z)), 5.63 (broad s, IH, NHZ), 7.06-7.72 (m, 16H, 15 aromatic H, NH amide), 8.09 (s, IH, NH indole). ¹³ C NMR (75 MHz, CDCl ₃ ) δ 28.6 (CH ₂ Trp), 56.1 (CHa), 67.2 (CH ₂ (Z)), 111.4, 118.8, 119.9, 120.1, 122.4, 123.4, 124.5, 127.1, 128.1, 128.3, 128.6, 128.9, 136, 136.2, 137.1 (20 aromatic C), 156 (CO carbamate), 169.7 (CO amide). Anal. Calcd. for C ₂₅ H ₂₃ N ₃ O ₃ : C, 72.62; H, 5.61; N, 10.16. Found: C, 72.75; H, 5.58; N, 9.95.

Z-TrP-NHCH ₂ Ph: Same procedure as above with Z-Trp-OH (2 g, 5.9 mmol), benzylamine (645 μL, 5.9 mmol), DCC (1.58 g, 7.65 mmol) and THF (88 mL). A white solid was obtained after precipitation in methanol/pentane (777 mg, 31%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 3.18 (dd, J = 14.3 Hz, J = 8.1 Hz, IH, CH ₂ ), 3.39 (dd, J = 14.3 Hz, J = 4.9 Hz, IH, CH ₂ ), 4.3 (m (AB), 2H, CH ₂ (Bn)), 4.53 (m, IH, CHa), 5.11 (s, 2H, CH ₂ (Z)), 5.52 (m, IH, NH), 5.90 (m, IH, NH), 6.91-7.70 (m, 15 aromatic H), 8.01 (s, IH, NH indole). ¹³ C NMR (75 MHz, CDCl ₃ ) δ 28.7 (CH ₂ Trp), 43.4 (CH ₂ ), 56.1 (CHa), 67 (CH ₂ (Z)), 111.2, 118.7, 119.7, 122.2, 123.2, 123.4, 127.2, 127.3, 127.6, 128, 128.2, 128.5, 136.1, 136.2, 137.5, 137.6 (20 aromatic C), 156 (CO carbamate), 171.2 (CO amide). Anal. Calcd. for C ₂₆ H ₂₅ N ₃ O ₃ , 0.75 H ₂ O: C, 70.81; H, 6.06; N, 9.53. Found: C, 70.84; H, 6.80; N, 9.21.

Z-Trp-NH(4-OH)Ph: Same procedure as above with Z-Trp-OH (0.5 g, 1.48 mmol), 4- aminophenol (162 mg, 1.48 mmol), DCC (396 mg, 1.92 mmol) and THF (20 mL). A white solid was isolated after purification by flash chromatography on silica gel (0-3% MeOH/CH ₂ Cl ₂ ) (493 mg, 77%). ¹ H NMR (300 MHz, MeOD) δ 3.25 (dd, J = 14.4 Hz, J=8

Hz, IH, CH ₂ ), 3.46 (dd, J=14.4 Hz, J= 5.3 Hz, IH, CH ₂ ), 4.53 (m, IH, CHa), 5.06 (m, 2H,

CH ₂ (Z)), 6.7 (d, J = 8.9 Hz, 2 aromatic H), 7 (t, J = 7 Hz, 1 aromatic H), 7.07-7.35 (m, 10 aromatic H), 7.6 (d, J = 7.8 Hz, 1 aromatic H). ¹³ C NMR (75 MHz, MeOD) δ 32.3 (CH ₂ Tip), 60.4 (CHa), 70.2 (CH ₂ (Z)), 113.3, 114.8, 118.6, 122, 122.4, 125, 126.5, 127.2, 131.3, 131.5, 133.5, 140.5, (20 aromatic C), 158.1 (CO carbamate), 175.2 (CO amide). Anal. Calcd. for C ₂₅ H ₂₃ N ₃ O ₄ , 0.25 H ₂ O: C, 69.19; H, 5.46; N, 9.68. Found: C, 69.34; H, 5.32; N, 9.61.

Z-Trp-NH(CH ₂ ) ₄ NHBoc: To a solution of Z-Trp-OH (1 g, 3 mmol) in DMF (16 mL) at 0 °C were added HOBt (611 mg, 4.52 mmol), EDC (636 mg, 3.32 mmol) and NEt ₃ (0.5 mL). The reaction was stirred for 30 min followed by drop wise addition of a solution of amine NH ₂ (CH ₂ ) ₄ NHBoc (prepared according to: Krapcho, A. P.; Kuell, C. S., Mono-protected diamines. N-tert-butoxycarbonyl-α,ω-alkanediamines from α,ω-alkanediamines., Synthetic Communications 1990, 20, (16), 2559-2564), (630 mg, 3.34 mmol) in DMF (4 mL) and NEt ₃ (0.5 mL). The reaction mixture was allowed to warm up to room temperature overnight and then concentrated. The resulting residue was diluted with CH ₂ Cl ₂ and successively washed with water, aqueous NaHCO ₃ . (1 M), aqueous KHSO ₄ (0.5 M) and brine. The organic phase was dried over Na ₂ SO ₄ and the solvent was removed in vacuo. The crude was triturated with CH ₂ Cl ₂ and a white solid was collected by filtration (773 mg). The mother liquor was concentrated down and triturated once more with CH ₂ Cl ₂ /pentane. A second batch was isolated as a white solid (482 mg, 82% overall yield). ¹ H NMR (300 MHz, CDCl ₃ ) δ 1.18 (m, 4H, 2 CH ₂ ), 1.48 (s, 9H, (CH ₃ ) ₃ ), 2.93-3.36 (m, 6H, CH ₂ Trp, 2 CH ₂ N), 4.5 (m, IH, CHa), 4.65 (m, IH, NHBoc), 5.12 (m, 2H, CH ₂ (Z)), 5.68 (m, 2 NH), 7.01 (s, 1 aromatic H), 7.11 (m, 1 aromatic H), 7.19 (t, J = 7 Hz, 1 aromatic H), 7.32 (m, 6 aromatic H), 7.7 (m, 1 aromatic H), 9 ( broad s, IH, NH indole). ¹³ C NMR (75 MHz, CDCl ₃ ) δ 26.5, 27.6, 29 (2 CH ₂ , CH ₂ Trp), 28.5 ((CHa) ₃ ), 39.1, 40.2 (CH ₂ NHCO, CH ₂ NHBoc), 55.8 (CHa), 67 (CH ₂ (Z)), 79.5 (C(CH ₃ ) ₃ ), 110.2, 111.4, 118.8, 119.7, 122.1, 123.5, 127.2, 128.1, 128.2, 128.5, 136.2, 136.3 (14 aromatic C), 156.4 (CO carbamate), 171.3 (CO amide). Anal. Calcd. for C ₂₈ H ₃₆ N ₄ O ₅ , 0.25 H ₂ O: C, 65.54; H, 7.16; N ₅ 10.91. Found: C, 65.78; H, 7.17; N, 10.63.

Trp-NHPh: A Schlenk flask charged with Z-Trp-NHPh (1.7 g, 4.11 mmol) and 10% Pd/C (437.4 mg) was flushed under H ₂ before adding MeOH/DMF (41 rnL, 1/1). The reaction mixture was stirred under atmosphere of H ₂ overnight followed by filtration through a pad of celite. The solvent was removed in vacuo and the crude was purified by flash column chromatography on silica gel (2-8% MeOH/CH ₂ Cl ₂ ). The amine was isolated as a yellow solid (841 mg, 73%). mp 114-116 ⁰ C. IH NMR (300 MHz, CDCl ₃ ) δ 1.89 (broad s, 2H, NH ₂ ), 3.06 (dd, J - 14.5 Hz, J = 8.8 Hz, IH, CH ₂ ), 3.55 (dd, J = 14.5 Hz, J = 3.9 Hz, IH, CH2), 3.89 (dd, J = 8.8 Hz, J = 3.8 Hz, IH, CHa), 7.06-7.41 (m, 7 aromatic H) ₃ 7.61 (d, J = 7.9 Hz, 2 aromatic H), 7.72 (d, J = 7.8 Hz, 1 aromatic H), 8,24 (broad s, IH, NH amide), 9.48 (s, IH, NH indole). 13C NMR (75 MHz, CDCl ₃ ) δ 30.5 (CH2 Tip), 56 (CHa), 111.4, 118.8, 119.5, 119.7, 122.3, 123.2, 124.1, 127.5, 129, 129.1, 136.5, 137.8 (14 aromatic C), 173.2 (CO amide).

Trp-NHCH ₂ Ph: Same procedure as above with Z-Trp-NHCH ₂ Ph (616 mg, 1.44 mmol), 10% Pd/C (153 mg) in MeOH/DMF (14.5 mL, 1/1). After purification by flash chromatography on silica gel (3-12% MeOH/CH ₂ Cl ₂ ), Trp-NHCH ₂ Ph was isolated as a pale yellow oil (351 mg, 83%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 2.98 (dd, J = 14.4 Hz, J = 8.7 Hz, IH, CH ₂ ), 3.44 (dd, J = 14.4 Hz, J = 4.1 Hz, IH, CH ₂ ), 3.80 (dd, J = 8.7 Hz, J - 4.2 Hz, IH, CHa), 4.46 (m, 2H, CH ₂ (Bn)), 7.06 (d, J = 2.1 Hz, 1 aromatic H), 7.13-7.36 (m, 7 aromatic H), 7.41 (d, J = 8 Hz, 1 aromatic H), 7.61 (m, IH, NH amide), 7.72 (d, J = 7.8 Hz, 1 aromatic H), 8.3 (broad s, IH, NH indole). ¹³ C NMR (75 MHz, MeOD) δ 30.5 (CH ₂ Tip), 42.7 (CH ₂ Ph), 55.4 (CHa), 109.3, 111, 118.1, 118.5, 121.2, 123.5, 126.8, 127, 127.1, 127.4, 128, 128.1, 136.8, 138 (14 aromatic C), 174.6 (CO amide).

Trp-NH(4-OH)Ph: Same procedure as above with Z-Trp-NHPhOH (208 nig, 0.48 mmol), 10% Pd/C (52 mg) in MeOH (5 mL). A light brown solid was obtained after precipitation in CH ₂ Cl ₂ MeOH (85 mg, 59%). ¹ H NMR (300 MHz, MeOD) δ 3.09 (dd, J = 14.1 Hz, J = 6.9 Hz, IH, CH ₂ ), 3.25 (dd, J = 14.1 Hz ₅ J = 6.2 Hz, IH ₅ CH ₂ ), 3.73 (t, J = 6.6 Hz, IH, CHa) ₅ 6.71 (d, J = 8.9 Hz ₅ 2 aromatic H), 7 (t, J = 8 Hz, 1 aromatic H), 7.1 (t, J = 8 Hz, 1 aromatic H), 7.12 (s ₅ 1 aromatic H), 7.21 (d, J = 8.9 Hz, 2 aromatic H), 7.35 (d, J = 8 Hz, 1 aromatic H), 7.64 (d, J = 8 Hz, 1 aromatic H). ¹³ C NMR (75 MHz, MeOD) δ 34.4 (CH ₂ Tip), 59.6 (CHa) ₅ 113.3, 114.9, 118.7, 122, 122.4, 125, 126.2, 127.4, 131.3, 133.5, 140.5, 158.1 (14 aromatic C) ₅ 177 (CO amide). Anal. Calcd. for C _n H ₁₇ N ₃ O ₂ , 0.25 H ₂ O: C, 67.86; H, 6.19; N ₅ 13.96. Found: C, 67.96; H, 5.97; N, 14.13.

Trp-NH(CH ₂ ) ₄ NHBoc: Same procedure as above with Z-Trp-NH(CH ₂ ) ₄ NHBoc (1.16 g, 2.28 mmol), 10% Pd/C (242 mg) in MeOH/DMF (10 mL, 1/1). After purification by flash chromatography on silica gel (4-25% MeOH/CH ₂ Cl ₂ ), Trp-NH(CH ₂ ) ₄ NHBoc was isolated as a white foam (701 mg, 82%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 1.39 (m, 4H ₅ 2 CH ₂ ), 1.46 (s, 9H ₅ (CHs) ₃ ), 3-3.35 (m, 6H, CH ₂ Trp, 2 CH ₂ N), 3.7 (m, IH, CHa), 4.71 (m, IH ₅ NHBoc), 7.05 (d, J = 2 Hz ₅ I aromatic H), 7.1 (t, J = 7.7 Hz, 1 aromatic H), 7.2 (m, 2H, 1 aromatic H, NH amide), 7.38 (d, J = 7.9 Hz, 1 aromatic H), 7.65 (d, J = 7.9 Hz, 1 aromatic H), 8.94 (broad s, IH, NH indole). ¹³ C NMR (75 MHz, CDCl ₃ ) δ 26.9, 27.6, 30.8 (2 CH ₂ , CH ₂ Trp), 28.5 ((CH ₃ ) ₃ ), 38.7, 40.3 (CH ₂ NHCO, CH ₂ NHBoc), 55.6 (CHa), 79.3 (C(CH ₃ ) ₃ ), 111.3, 111.4, 118.9, 119.5, 122, 123.3, 127.6, 136.4 (8 aromatic C), 156.2 (CO carbamate), 174.8 (CO amide).

C) Preparation of oxotryptophane derivatives:

L-Z-Trp[O]-OH: To a solution of L-H-Trp[O] -OH (2.55 g, 11.57 mmol, prepared according to : Labroo, R. B.; Cohen, L. A., Preparative separation of the diastereoisomers of dioxindolyl-L-alanine and assignment of stereochemistry at C-3. J. Org. Chem. 1990, 55, (16), 4901-4904) in DMF (11.5 mL) was added ZOSu (2.89 g, 11.6 mmol) and then NEt ₃ (1.63 mL, 11.7 mmol). The solution was stirred for 4 h at room temperature and then was concentrated in vacuo. The resulting residue was triturated in 5% aqueous KHSO4 (20 mL) and the resulting mixture was extracted with CH ₂ Cl ₂ (S X 25 mL). After drying of the organic phases on MgSO4 and concentration in vacuo, L-Z-Trp[O] -OH was obtained as a beige solid (2.28 g, 55 %). IH NMR(300 MHz, DMSO-d6)(50/50 mixture of two diastereomers) δ 1.91- 2.27 (m, 2H, CH2 Trp), 3.42 (m, IH, CH oxindole), 4.38 and 4.52 (two m, IH, CHa dia 1 or dia 2), 5,05 (s, 2H, CH2(Z)), 6.83 and 6.95 (two t, J = 10Hz and J = 7.8 Hz, IH, aromatic H of dia 1 or dia 2), 7.17 and 7.26 (two t, J = 5.2 Hz and J = 8.4 Hz, IH, aromatic H of dia 1 or dia 2), 7.32-7.44 (m, 7H, aromatic H), 7.75 and 7.88 (two d, J - 8 Hz and J = 8.7 Hz, IH, NH(Z) dia 1 or dia 2), 10.41 and 10.43 (two s, IH, NH oxindole dia 1 or dia 2), 12.52 (broad s, IH, acidic H). 13C NMR (75 MHz, DMSO-d6) (mixture of two diastereomers) δ 32.3 (CH2), 41.5 and 42.1 (CH), 51.3 and 51.4 (CH), 65.4 and 65.5 (CH2), 109.2 and 109.4 (CH), 121.2 and 121.32 (CH), 123.9 and 124.34 (CH), 127.6, 127.7 127.8, 128.3 (4 CH), 128.8 and 129.3 (C), 136.9 (C), 142.4 and 142.6 (C), 156.1 and 156.2 (C), 173.3 and 173.7 (C), 178.4 and 178.7 (C). Anal. Calcd. for C ₁₉ H ₁₈ N ₂ O ₅ , 1 H ₂ O : C, 61.28 ; H, 5.41; N, 7.52. Found: C, 61.65; H, 4.91; N, 7.50.

L-Z-Trp[O]-NHPh: To a solution of crude L-Z-Trp[O] -OH (2.22 g, 6.26 mmol) in DME (12.5 mL) was added at O ⁰ C N-hydroxysuccinimide (0.757 g, 6.58 mmol) and dicyclohexylcarbodiimide (1.36 g, 6.58 mmol). After 15 min at 0 °C, the mixture was stirred at room temperature overnight. The white solid was filtered and washed by DME (3 X 5 mL).

The filtrate was concentrated in vacuo and the residue was dissolved in CH ₂ Cl ₂ (50 mL). The resulting solution was washed by water (3 X 10 mL), dried over Na ₂ Sθ4 and concentrated in vacuo to afford L-Z-Trp-OSu as a light yellow solid (2.56 g, 90 %) which was used without

further purification. To a solution of this crude product in DME (9 mL) was added freshly distilled aniline (0.62 mL, 6.80 mmol). After stirring at room temperature overnight, the solvent was evaporated in vacuo and the solid residue was suspended in 5 % aqueous KHSO ₄ (15 mL). After filtration, washing of the solid with water (2 X 10 mL), suspension of the solid in boiling 95 % EtOH (20 mL) Z-Trp[O] -NHPh was obtained as a very fine white powder (1.29 g, 48 % from L-Z-Trp[O] -OH). ¹ H NMR (300 MHz, DMSO-d ₆ ), (one diastereomer, which slowly underwent isomerization to a mixture of two diastereomers): δ 2.17 (m, 2H, CH ₂ Trp), 3.50 (m, IH, CH oxindole), 4.67 (m, IH, CHa), 5.08 (broad s, 2H, CH ₂ (Z)), 6.86 (d, J = 7.5 Hz, IH, NH Z), 6.98 (t, J = 7.6 Hz, IH), 7.09 (t, J = 7.2 Hz, IH), 7.20 (t, J = 7.6 Hz, IH), 7.35-7.73 (m, 15H, 14 aromatic H and 1 NH(Z)) , 10.1 (s, IH, NH) , 10.48 (s, IH, NH). 13C NMR (75 MHz, DMSO-d ₆ ) (mixture of two diastereomers) δ 32.8 (CH ₂ Trp), 41.7 and 42.0 (CHy), 53.1 (CHa), 65.4 and 65.5 (CH ₂ Z), 109.2 and 109.4 (CH), 119.4 and 119.5 (CH), 121.3 (CH), 123.4 (CH), 124.0 and 124.7 (CH), 127.6, 127.7, 127.8, 128.3. and 128.6 (CH), 128.8 and 129.3 (C), 136.8 and 136.9 (C), 138.6 and 138.8 (C), 142.4 and 142.6 (C) ₅ 155.8 and 156.2 (C), 170.3 and 170.6 (C), 178.5 and 178.6 (C). Anal. Calcd. for C ₂₅ H ₂₃ N ₃ O ₄ , 0.5 H ₂ O : C, 68.48; H, 5.52; N, 9.58. Found: C, 68.47; H, 5.20; N, 9.40.

L-Z-Trp [O]-NHCH ₂ Ph: Same procedure as above starting from Z-Trp-OSu (1.54 g, 3.29 mmol) and benzylamine (0.43 mL, 3.95 mmol) afforded L-Z-Trp[O]-NHCH ₂ Ph after recristallization in EtOH (0.775 g, 53%) as a white solid. ¹ H NMR (300 MHz, DMSO-d ₆ ), (60/40 mixture of two diastereomers in equilibrium): δ 1.88 and 2.11 (two m, 2H, CH ₂ Trp), 3.45 (m, IH, CH oxindole), 4.28 and 4.30 (two s, 2H, NCH ₂ Ph), 4.50 (m, IH, CHa), 5.06 and 5.08 (two s, 2H, CH ₂ (Z)), 6.85 (t, J = 7.6 Hz, IH, aromatic H), 6.97 (t, J = 7.2 Hz, IH, aromatic H), 7.30 (m, 12H, 12 aromatic H), 7.65 and 7.87 (two d, J = 8.9 Hz, IH, NHZ), 8.53 (m, IH, NH Bn), 10.46 and 10.48 (two s, IH, NH oxindole). 13C NMR (75 MHz, DMSO- d ₆ ) (mixture of two diastereomers) δ 33.0 and 33.2 (CH ₂ β Trp), 42.1 (CHy), 42.2 (CH ₂ Bn), 52.5 and 52.6 (CHa), 65.5 and 65.6 (CH ₂ Z), 109.2 and 109.4 (CH), 121.2 and 121.3 (CH), 124.0, 124.6, 126.6, 126.7, 126.9, 127.0, 127.1, 127.3, 127.4, 127.6, 127.7, 127.8, 128.1, 128.2 (aromatic CH) ₅ 128.3 and 129.0 (C), 136.9 (C), 139.2 and 139.4 (C), 142.4 and 142.5 (C), 155.8 and 156.2 (C), 171.3 and 171.5 (C), 178.6 and 178.7 (C). Anal. Calcd. for C ₂₆ H ₂₅ N ₃ O ₄ , 0.5 H ₂ O : C, 69.01; H, 5.79; N, 9.29. Found: C, 69.00; H, 5.76; N, 9.14.

HCl, L-Trp [O]-OMe : was prepared according to : Von Nussbaum, F.; Danishefsky, S. J. A rapid total synthesis of spirotryprostatin B: proof of its relative and absolute stereochemistry. Angew. Chem. Int .Ed. 2000, 39(12), 2175-2178.

D) Preparation of dipep tides:

Dipeptides were prepared using conventional peptide synthesis and were obtained according to Berthelot, A.; Piguel, S.; Le Dour, G.; Vidal, J., Synthesis of macrocyclic peptide analogues of proteasome inhibitor TMC-95 A. J. Org. Chem. 2003, 68, (25), 9835-9838.

N-Boc-Tyr(Bn)-AIa-OMe. described in the above article

N-Boc-Tyr(Bn)-Leu-OMe: described in the above article

N-Boc-Tyr(Bn)-Asn-OMe: described in the above article

N-Boc~Tyr(Me)~AIa-OMe:. This compound was synthesized as described above from N- Boc-Tyr(Me)-OSu (1 g, 2.55 mmol), HCl, AIa-OMe (320.5 mg, 2.3 mmol) and NEt ₃ (323 μL, 2.3 mmol). The dipeptide was obtained as a white solid (900 mg, 93%) and was used in the next step without further purification. [α] _D ²⁰ -9.02 (c 1, MeOH). ¹ H NMR (300 MHz, CDCl ₃ ) δ 1.34 (d, J = 7.1 Hz, 3H, CH ₃ (AIa)), 1.46 (s, 9H, 3 CH ₃ (Boc)), 3.02 (m, 2H, CH ₂ (Tyr)), 3.73 (s, 3H, OCH ₃ ), 3.8 (s, 3H, OCH ₃ ), 4.31 (m, IH, CH (Tyr)), 4.52 (m, IH, CH (AIa)), 4.97 (broad s, IH, NH (Boc)), 6.41 (d, J = 7.1 Hz, IH, NH), 6.86 (d, J = 8.6 Hz, 2H, H3) ; 7.14 (d, J = 8.7 Hz, 2H, H2). HRMS (ESI) calcd for C ₁₉ H ₂₈ N ₂ O ₆ Na [(M+Na) ⁺ ] 403.1845 , found 403.1847. These data are in agreement with those of Boger, D. L.; Zhou, J. N-Desmethyl Derivatives of Deoxybouvardin and RA-VII: Synthesis and Evaluation. J. Am. Chem. Soc. 1995, 117(28), 7364-78.

N-Boc-Tyr(Me)-Leu-OMe. This compound was synthesized as described above from N- Boc-Tyr(Me)-OSu (1 g, 2.55 mmol), Leu-OMe.HCl (418 mg, 2.3 mmol) and NEt ₃ (323 μL, 2.3 mmol). The dipeptide was obtained as a white solid (952 mg, 98%) and was used in the next step without further purification. ¹ H NMR (300 MHz, CDCl ₃ ) δ 0.81 (m, 6H, 2 CH ₃ (Leu)), 1.31 (s, 9H, 3 CH ₃ (Boc)), 1.49 (m, 3H, CH and CH ₂ (Leu)), 2.9 (m, 2H, CH ₂ (Tyr)) ; 3.6 (s, 3H, OCH ₃ ), 3.66 (s, 3H, OCH ₃ ), 4.27 (m, IH, CH (Tyr)), 4.48 (m, IH, CHa (Leu)), 5.29 (m, IH, NH (Boc)), 6.71 (d, J = 8.4 Hz, 2H, H3), 6.75 (broad s, IH, NH) ₅ 7.03 (d, J = 8.4 Hz, 2H, H2). ¹³ C NMR (50 MHz, CDCl ₃ ) δ 21.8 (CH ₃ (Leu)), 22.7 (CH ₃ (Leu)), 24.6 (CH (Leu)), 28.2 (3 CH ₃ (Boc)), 37.4 (CH ₂ (Tyr)), 41.2 (CH ₂ (Leu)), 50.7 (CHa (Leu)), 52 (OCH ₃ ), 55 (OCH ₃ ), 55.6 (CH (Tyr)), 79.7 (C (Boc)), 113.8 (C3), 128.7 (Cl), 130.3 (C2), 155.4 (C4), 158.4 (CO (Boc)), 171.4 (CO amide), 172.9 (CO ester). HRMS (ESI) calcd for C ₂₂ H ₃₄ N ₂ O ₆ Na [(M+Na) ⁺ ] 445.2315, found 445.2319.

N-Boc-Tyr(Me)-Asn-OMe: This compound was synthesized as described above from N- Boc-Tyr(Me)-OSu (1 g, 2.55 mmol), HCl, Asn-OMe (417 nig, 2.3 mmol) and NEt ₃ (323 μL, 2.3 mmol). The dipeptide was obtained as a white solid (653 mg, 67%) which was used in the next step without further purification. ¹ H NMR (300 MHz, CDCl ₃ ) δ 1.27 (s, 9H, 3 CH ₃ (Boc)), 2.72-2.98 (m, 4H, 2 CH ₂ ), 3.62 (s, 3H, OCH ₃ ), 3.65 (s, 3H, OCH ₃ ), 4.4 (m, IH, CH (Tyr)), 4.7 (m, IH, CH, (Asn)), 5.46 (broad s, IH, NH (Boc)), 6.27 (broad s, IH, NH ₂ ), 6.5 (broad s, IH, NH ₂ ), 6.72 (d, J = 8.5 Hz, H3), 7.03 (d, J = 7.8 Hz, H2), 7.8 (broad s, IH, NH). ¹³ C NMR (300 MHz, CDCl ₃ ) δ 28.2 (3 CH ₃ (Boc)), 37 (CH ₂ (Asn)), 37.6 (CH ₂ (Tyr)), 49 (CH (Asn)), 57.6 (OCH ₃ ), 55.1 (OCH ₃ ), 55.5 (CH (Tyr)), 79.8 (C (Boc)), 113.8 (C3), 128.6 (Cl), 130.4 (C2), 155.5 (C4), 158.4 (CO (Boc)), 171.6 (CO), 172 (CO) ₅ 172.7 (CO). HRMS (ESI) calcd for C ₂₀ H ₂₉ N ₃ O ₇ Na [(M+Na) ⁺ ] 446.1903, found 446.1896.

442,5

N~Boc-Tyr(Bn)-Gly-OMe: Same procedure as for N-Boc-Tyr(Bn)- AIa-OMe with N-Boc- Tyr(Bn)-OSu (1 g, 2.13 mmol), Gly-OMe.HCl (268 mg, 2.14 mmol) and NEt ₃ (0.3 niL, 2.16 mmol) in DMF (4 mL). The dipeptide was obtained as a white solid (946 mg, 100%) which was used in the next step without further purification. Recristallization of dipeptide from hot iso-propanol afforded an analytical sample, mp 118-120 °C (litt 118-120 ⁰ C Flouret, G. R.; Arnold, W. H.; Cole, J. W.; Morgan, R. L.; White, W. F.; Hedlund, M. T.; Rippel, R. H. J. Med. Chem. 1973, 16(4), 369-73.). ¹ H NMR (300 MHz, CDCl ₃ ) δ 1.42 (s, 9H, (CH ₃ ) ₃ ), 3.05 (m, 2H, CH ₂ Tyr), 3.75 (s, 3H, CO ₂ Me), 3.95 (dd, J = 18.1 Hz, J = 5 Hz, IH, CH ₂ GIy), 4.05 (dd, J = 18.2 Hz, J = 5.4 Hz, IH, CH ₂ GIy), 4.38 (m, IH, CHa), 5.05 (broad s, 3H, NHBoc, CH ₂ (Bn)), 6.47 (m, IH, NH amide), 6.93 (d, J = 8.5 Hz, 2 aromatic H), 7.15 (d, J = 8.5 Hz, 2 aromatic H), 7.32-7.45 (m, 5 aromatic H). ¹³ C NMR (75 MHz, CDCl ₃ ) δ 28.3 ((CH ₃ ) ₃ ), 37.5 (CH ₂ Tyr), 41.2 (CH ₂ GIy), 52.3 (OCH ₃ ), 55.7 (CHa), 70 (CH ₂ (Bn)), 80.2 (C(CH ₃ ) ₃ ), 114.9, 127.5, 128, 128.3, 128.6, 128.9, 130.4, 137.1 (11 aromatic C), 155.5, 157.8 (Car-O, CO carbamate), 170, 171.9 (CO amide, CO ester).

N-Boc-Tyr(Bn)-Arg(Z) ₂ -OH: Same procedure as above with N-Boc-Tyr(Bn)-Osu (837 mg, 1.78 mmol), H-Arg(Z) ₂ -OH (791 mg, 1.78 mmol) in DMF (10 mL). The reaction mixture was stirred for 4 days followed by usual work-up. The dipeptide was isolated after precipitation in CH ₂ Cl ₂ /pentane (972 mg, 68%). ¹ H NMR (300 MHz, CDCI ₃ ) δ 1.37 (s, 9H, (CH ₃ ) ₃ ), 1.6 (m, 2H, CH ₂ Arg), 1.79 (m, 2H, CH ₂ Arg), 2.85-3 (m, 2H, CH ₂ Tyr), 3.92 (m, 2H, CH ₂ NZ), 4.29 (m, IH, CHa), 4.48 (m, IH, CHa), 4.99-5.23 (m, 8H, 3 CH ₂ , 2 NH) ₅ 6.85 (d, J = 8.4 Hz, 2 aromatic H), 7.02 (d, J = 8.4 Hz, 2 aromatic H), 7.1 (m, 1 NH), 7.38 (m, 15 aromatic H), 9.43 (m, 1 NH). ¹³ C NMR (50 MHz, CDCI ₃ ) δ 25.1, 25.8 (2 CH ₂ Arg), 28.6 ((CH ₃ ) ₃ ), 37.7 (CH ₂ Tyr), 44.5 (CH ₂ NHZ), 53, 56 (CHa Tyr, Arg), 67.5, 69.4, 70.3 (2 CH ₂ (Z), CH ₂ (Bn)), 80.6 (C(CH ₃ ) ₃ ), 115.3, 127.8, 128.4, 128.7, 128.9, 129.2, 130.7, 135, 137, 137.4 (23 aromatic C), 156, 156.1, 158.1, 161, 163.9 (Car-O, 3 CO carbamate, CO imine), 172.4, 174.8 (CO amide, CO acid).

N-Boc-Tyr(Bn)-Lys(Boc)-OH: Same procedure as above with N-Boc-Tyr(Bn)-OSu (1 g, 2.13 mmol), H-Lys(Boc)-OH (550 mg, 2.23 mmol) and a few drops Of NEt ₃ in DMF (3.5 mL). The dipeptide was isolated after precipitation in CH ₂ Cl ₂ /pentane (946 mg, 74%). mp 168-170 ⁰ C. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.2-1.35 (m, 4H, 2 CH ₂ Lys), 1.29 (s, 9H, (CH ₃ ) ₃ ), 1.35 (s, 9H, (CRi) ₃ ), 1.6 (m, 2H, CH ₂ Lys), 2.55-3.05 (m, 4H, CH ₂ Tyr, CH ₂ Lys), 3.63 (m, IH, CHa), 3.94 (m, IH, CHa), 5.05 (s, 2H, CH ₂ (Bn)), 6.70 (m, IH, NHBoc), 6.88 (d, J = 8.2 Hz ₅ 2 aromatic H) ₅ 7.14 (d, J - 8.2 Hz ₅ 2 aromatic H) ₅ 7.31-7.52 (m, 5 aromatic H). ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 22.6, 26.8 (2 CH ₂ Lys), 28.6, 28.7 (2 (CH ₃ ) ₃ ), 30 (CH ₂ Lys), 32.6 (CH ₂ NHBoc), 36.8 (CH ₂ Tyr), 54.4, 56.9 (CHa Tyr, Lys), 69.5 (CH ₂ (Bn)), 77.8, 78.6 (2 C(CHa) ₃ ), 114.7, 128, 128.2, 128.8, 130.6, 131.1, 137.6 (11 aromatic C), 155.7, 156, 157.2 (Car-O, 2 CO carbamate), 170.7, 174.7 (CO amide, CO acid).

N-Boc-Tyr(Bn)-Gly-OH:A solution Of N-BoC-TyT(Bn)-GIy-OMe (500 sag, 1.13 mmol) in THF (1.4 mL) was treated with aqueous LiOH (1 M, 1.4 niL, 1.4 mmol) at O ⁰ C for lh30. The reaction mixture was quenched with aqueous 4 N HCl. The aqueous phase was extracted with CH ₂ Cl ₂ . The combined organic phases were dried over Na ₂ SO ₄ and concentrated in vacuo to give crude carboxylic acid (435 mg, 89%) which was taken in the next step without further purification, mp 157-159 °C. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.29 (s, 9H ₅ (CH ₃ ) ₃ ), 2.65 (dd, J = 13.7 Hz, J = 10.5 Hz, IH, CH ₂ Tyr), 2.93 (dd, J = 13.7 Hz, J = 3.5 Hz, IH, CH ₂ Tyr), 3.77 (m, 2H, CH ₂ GIy), 4.13 (m, IH, CHa Tyr), 5.05 (m, 2H, CH ₂ (Bn)), 6.86 (d, J = 8.9 Hz, IH, NHBoc), 6.93 (d, J = 8.5 Hz, 2 aromatic H), 7.18 (d, J = 8.5 Hz, 2 aromatic H), 7.25-7.45 (m, 5 aromatic H), 8.19 (m, IH, NH amide). ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 26.2 ((CH ₃ ) ₃ ), 34.7 (CH ₂ Tyr), 38.8 (CH ₂ GIy), 53.9 (CHa Tyr), 67.2 (CH ₂ (Bn)), 76 (C(CH ₃ ) ₃ ), 112.4, 125.7, 125.8, 126.5, 128.3, 128.5, 135.4 (11 aromatic C), 153.3, 154.9 (Car-0, CO carbamate), 169.3, 170.2 (CO acid, CO amide).

4 44422,,5

N-Boc-Tyr(Bn)- AIa-OH: Same procedure as above with N-Boc-Tyr(Bn)-Ala-OMe (1.55 g, 3.39 mmol), aqueous LiOH (1 M, 4 mL, 4 mmol) in THF (4 mL). A white solid was obtained (1.25 g, 83%) which was used in the next step without further purification. ¹ H NMR (200 MHz, CDCI ₃ ) δ 1.41 (s large, 12H, CH ₃ , (CH ₃ ) ₃ ), 3.01 (m, 2H, CH ₂ ), 4.38 (m, IH, CHa), 4.53 (m, IH, CHa), 5.04 (s, 2H, CH ₂ (Bn)), 5.16 (broad s, IH, NHBoc), 6.68 (m, IH, NH amide), 6.91 (d, J = 8.6 Hz, 2 aromatic H), 7.12 (d, J = 8.6 Hz, 2 aromatic H), 7.33-7.44 (m, 5 aromatic H). ¹³ C NMR (75 MHz, CDCI ₃ ) δ 18 (CH ₃ ), 28.2 ((CHa) ₃ ), 37.5 (CH ₂ Tyr), 48.2 (CHa Ala), 55.6 (CHa Tyr), 70 (CH ₂ (Bn)), 80.5 (C(CH ₃ ) ₃ ), 115, 127.4, 128, 128.5, 128.6, 130.4, 137 (11 aromatic C), 155.8, 157.8 (Car-O, CO carbamate), 171.6, 175.5 (CO acid, CO amide).

N-Boc-Tyr(Bn)-Leu-OH: Same procedure as above with N-Boc-Tyr(Bn)-Leu-OMe (1.034 g, 2.073 mmol), aqueous LiOH (1 M, 2.2 mL, 2.2 mmol) in THF (15 mL). A white solid was obtained (1.035 g, 100%) which was used in the next step without further purification. ¹ H NMR (300 MHz, CDCI ₃ ) δ 0.90 (d, J = 5 Hz, IH, Me ₂ Leu), 1.40 (s, 9H, (CHs) ₃ ), 1-60 (m, 3H, CH ₂ -CH Leu), 3.00 (m, 2H, CH ₂ ), 4.29 (m, IH, CHa), 4.44 (m, IH, CHa), 4.88 (s, 2H, CH ₂ (Bn)), 5.29 (broad s, IH, NHBoc), 6.70 (m, IH, NH amide), 6.91 (d, J = 5 Hz ₅ 2 aromatic H), 7.12 (d, J = 5 Hz, 2 aromatic H), 7.33-7.44 (m, 5 aromatic H). ³ C NMR (75 MHz, CDCl ₃ ) δ 21.8, 22.9, 24.7 (CH-Me ₂ Leu), 28.2 ((CHa) ₃ ), 37.0 CH ₂ Leu), 41.1 (CH ₂ Tyr), 51.6 (CHa Leu), 55.7 (CHa Tyr), 70 (CH ₂ (Bn)), 80.4 (C(CH ₃ ) ₃ ), 114.9, 127.5, 127.9, 128.5, 128.9, 130.5, 137.0 (11 aromatic C), 155.8, 157.8 (Car-O, CO carbamate), 172.0, 176.8 (CO acid, CO amide).

N-Boc-Tyr(Bn)-Asn-OH: Same procedure as above with N-Boc-Tyr(Bn)-Asn-OMe (1.035 g, 2.072 mmol), aqueous LiOH (1 M, 2.2 mL, 2.2 mmol) in THF (15 mL). A beige solid was obtained (1.03 g, 100%) which was used in the next step without further purification. ¹ H NMR (300 MHz, CD ₃ OD) δ 1.35 (s, 9H, (CH ₃ ) ₃ ), 2.73-3.13 (m, 4H, CH ₂ ), 4.29 (m, IH, CHa), 4.72 (m, IH, CHa), 5.06 (s, 2H, CH ₂ (Bn)), 6.92 (d, J = 5 Hz, 2 aromatic H), 7.17 (d, J = 5 Hz, 2 aromatic H), 7.33-7.44 (m, 5 aromatic H). ¹³ C NMR (75 MHz, CD ₃ OD) δ 28.7 ((CH ₃ ) ₃ ), 37.7, 38.3 (CH ₂ Tyr and Asn), 50.4 (CHa Asn), 57.5 (CHa Tyr), 71 (CH ₂ (Bn)), 80.7 (C(CH ₃ ) ₃ ), 116.0, 128.5, 128.8, 129.5, 130.9, 131.5, 131.8, 138.8 (11 aromatic C), 157.6,, 159.2 (Car-O, CO carbamate), 174.2, 174.4, 175.0 (CO acid, CO amide).

II) Preparation of halogen ated tripeptides: compounds IV-Ia

Bromo, iodo tripeptides were prepared according to Berthelot, A.; Piguel, S.; Le Dour, G.; Vidal, J., Synthesis of macrocyclic peptide analogues of proteasome inhibitor TMC-95A. J. Org. Chem. 2003, 68, (25), 9835-9838.

N-Boc-3-iodo-Tyr(Bn)-AIa-7-bromo-Trp-OMe: compound A248 described in the above article.

N-Boc-3-iodo-Tyr(Bn)-Leu-7-bromo-Trp-OMe: compound A268 described in the above article.

N-Boc-3-iodo-Tyr(Bn)-Leu-7-bromo-Trp-OEt : compound Al 74. The peptide coupling was performed in CH ₂ Cl ₂ (0.5 mL) using 7-bromo-Trp-OEt (15 mg, 0.048 mmol), crude N- Boc-3-iodo-Tyr(Bn)-Leu-OH (31 mg, 0.05 mmol), EDC (11 mg, 0.053 mmol), HOBt (8 mg, 0.053 mmol) and NEt ₃ (15 μL, 0.1 mmol). The residue was purified by preparative TLC on silica gel (2% MeOH/CH ₂ Cl ₂ ) and subjected to crystallization with Et ₂ O/pentane to afford tripeptide N-Boc-3-iodo-Tyr(Bn)-Leu-7-bromo-Trp-OEt (29 mg, 67%) as a white amorphous solid. ¹ H NMR (300 MHz, CDCl ₃ , COSY) δ 0.81 (d, J = 6.2 Hz, 3H, CH ₃ Leu), 0.87 (d, J = 6.2 Hz, 3H, CH ₃ Leu), 1.2 (t, J = 7.1 Hz, 3H, CH ₃ (Et)), 1.41 (s, 9H, (CH ₃ ) ₃ ), 1.52-1.61 (m, 3H, CH ₃ CH ₂ Leu), 2.91 (m, 2H, CH ₂ Tyr), 3.26 (m, 2H, CH ₂ Tip), 4.09 (m, 2H, OCH ₂ (Et)), 4.28 (m, IH, CHa Tyr), 4.47 (m, IH, CHa Leu), 4.85 (m, IH, CHa Tip), 4.90 (d, IH, J = 7.9 Hz, NHBoc), 5.04 (s, 2H, CH ₂ Bn), 6.51 (m, IH, NH), 6.71 (d, J = 8.4, IH, H5 Tyr), 6.75 (d, J = 8.1 Hz, IH, NH), 6.95 (t, J = 7.7 Hz, IH, H5 Tip), 7.04 (m, 2H, aromatic H), 7.25-7.46 (m, 7H, aromatic H), 7.59 (d, J = 2 Hz, IH, H2 Tyr), 8.57 (broad s, IH, NH ind). ¹³ C NMR (75 MHz, CDCl ₃ ) 14.1 (CH ₃ Et), 22.2 (CH ₃ Leu), 22.9 (CH ₃ Leu), 24.7 (CH Leu), 27.7 (CH ₂ Tip), 28.3 ((CHs) ₃ ), 36.1 (CH ₂ Tyr), 40.9 (CH ₂ Leu), 51.7 (CHa Leu), 52.8 (CHa Tip), 55.8 (CHa Tyr), 61.7 (CH ₂ Et), 70.9 (CH ₂ Bn), 80.8 (C(CH ₃ ) ₃ ), 87 (C3 Tyr), 104.9 (C7 Tip), 111 (C), 112.7 (CH), 117.9 (CH), 120.7 (CH), 124.1 (CH), 124.5 (CH), 127 (CH), 128 (CH), 128.6 (CH), 128.8 (C), 130.3 (CH), 130.9 (C), 134.8 (C), 136.5 (C), 140.2 (CH), 155.8 (C4 Tyr), 156.4 (CO Boc), 177.1, 171.2, 171.5 (2 CO amide, CO ester). Anal. Calcd. for C ₄₀ H ₄₈ N ₄ O ₇ BrI: C, 53.17; H, 5.36; N, 6.20. Found: C, 53.06; H, 5.34; N, 6.10.

N-Boc-3-iodo-Tyr(Me)-AIa-7-bromo-Trp-OMe : compound A385: The peptide coupling was performed in CH ₂ Cl ₂ (1.5 mL) using 7-bromo-Trp-OMe (100 mg, 0.3 mmol), crude N- Boc-3-iodo-Tyr(Me)- AIa-OH (148 mg, 0.3 mmol), EDC (63 mg, 0.33 mmol), HOBt (45 mg, 0.33 mmol) and NEt ₃ (93 μL, 0.66 mmol). The residue was subjected to flash chromatography on silica gel (2% MeOH/CH ₂ Cl ₂ ) to afford tripeptide N-Boc-3-iodo- Tyr(Me)-Ala-7-bromo-Trp-OMe (204 mg, 88%) as a white amorphous solid. Rf 0.3 . ¹ H NMR (300 MHz, CDCl ₃ ) δ 1.31 (d, J = 7 Hz, 3H, CH ₃ ), 1.4 (s, 9H, (CH ₃ ) ₃ ), 2.83 (m, CH2 Tyr), 3.3 (m, 2H, CH2 Tip), 3.68 (s, 3H, OCH ₃ ), 3.8 (s, 3H, OCH ₃ ), 4.35 (m, IH, CH Tyr), 4.62 (q, J = 7.2 Hz, IH, CH Ala), 4.92 (m, IH, CH Tip), 5.22 (broad d, J = 7.6 Hz ₅ IH,

NHBoc), 6.67 (d, J = 8.4 Hz, IH, H5 Tyr), 6.96 (t, J = 7.7 Hz, IH, H5 Tip), 7.03-7.08 (m, 2 aromatic H), 7.27 (d, J = 7.7 Hz, 1 aromatic H Tip), 7.46 (d, J = 7.7 Hz, 1 aromatic H Trp),7.52 (d, J = 1.3 Hz, IH, H2 Tyr), 8.83 (s, IH, NHind). ¹³ C NMR (75 MHz, CDCl ₃ ) δ 18.5 (CH ₃ Ala), 26.4 (CH ₂ Tip), 28.3 ((CH ₃ ) ₃ ), 35.4 (CH ₂ Tyr), 48.9 (CH Ala), 52.5 (OCH ₃ ), 52.9 (CH Tip), 55.5 (CH Tyr), 56.3 (OCH ₃ ), 80.6 (C(CHa) ₃ ), 86 (C3 Tyr), 104.9 (C7 Tip), 110.8 (C3 Tip), 110.9 (CH), 117.7 (CH), 120.7 (CH), 124.2 (CH), 124.4 (CH), 128.7 (C), 130.3 (CH), 130.6 (CH), 134.8 (C), 140.1 (CH(2) Tyr), 155.6 (C4 Tyr), 157.1 (CO Boc), 171.3 , 171.8, 171.9 (2 CO amide, CO ester). HRMS (ESI) calcd for C ₃₀ H ₃₆ N ₄ O ₇ ⁷⁹ BrINa [M+Na] ⁺ 793.0710, found 793.0709.

N-Boc-3-iodo-Tyr(Me)-Leu-7-bromo-Trp-OMe : compound A363 described in the above article.

N-Boc-3-iodo-Tyr(Bn)-Asn-7-bromo-Trp-OMe : compound SP274 described in the above article

N-Boc-3-iodo-Tyr(Bn)-Ala-7-bromo-Trp-OH : compound A215. A solution of N-3- iodo-Tyr(Bn)-Ala-7-bromo-Trp-Ome (85.5 mg, 0.100 mmol) in THF (0.4 mL) was treated at 0 ⁰ C by 1 M aqueous NaOH (0.12 mL, 0.12 mmol). After 5 hours at room temperature, 1 M aqueous HCl was added (0.36 mL, 0.36 mmol). The resulting mixture was diluted by water and extracted by CH ₂ Cl ₂ (3 X 10 mL). After drying of the organic phase over Na ₂ SO ₄ and evaporation of the solvent, the residue was subjected to flash chromatography on silica gel (2% MeOH/CH ₂ Cl ₂ ) to afford remaining N-Boc-3-iodo-Tyr(Me)-Ala-7-bromo-Trp-OCH ₃ (10.13 mg, 12%) and N-Boc-3-iodo-Tyr(Me)-Ala-7-bromo-Trp-OH (58.3 mg, 70%, corrected yield 82%) as a white amorphous solid. ¹ H NMR (200 MHz, CDCl ₃ ) δ 0.9 (d, J = 6.4 Hz, 3H, CH ₃ ), 1.42 (s, 9H, (CHa) ₃ ), 2.87 (m, CH ₂ Tyr), 3.32 (m, 2H, CH ₂ Tip), 4.40 (m, IH, CH Tyr), 4.52 (m, IH CH Ala), 4.85 (m, IH, CH Tip), 5.06 (s, 2H, CH ₂ O), 5.16 (broad d, J = 7.6 Hz, IH, NHBoc), 6.71 (d, J - 8 Hz, IH, H5 Tyr), 6.91 - 7.58 (m, 14 H, aromatic H and NH), 8.68 (s, IH, NHind). HRMS (ESI) calcd for C ₃₅ H ₃₈ ⁷⁹ BrIN ₄ O ₇ Na [(M+Na) ⁺ ] 855.0866, found 855.0896.

N-Boc-3-iodo-Tyr(Me)-Leu-7-bromo-Trp-OEt : compound A340. The peptide coupling was performed in CH ₂ Cl ₂ (3.7 mL) using 7-bromo-Trp-OEt (294 mg, 0.75 mmol), crude N- Boc-3-iodo-Tyr(Me)-Leu-OH (400 mg, 0.75 mmol), EDC (172 mg, 0.9 mmol), HOBt (121 mg, 0.9 mmol) and NEt ₃ (230 μL, 1.65 mmol). The residue was subjected to flash chromatography on silica gel (5% MeOH/CH ₂ Cl ₂ ) to afford tripeptide N-Boc-3-iodo- Tyr(Me)-Leu-7-bromo-Trp-OEt (334 mg, 54%) as a white amorphous solid. Rf 0.6 (5% MeOHZCH ₂ Cl ₂ ), ¹ H NMR (300 MHz, CDCl ₃ ) δ 0.89 (d, J = 6.0 Hz, 3H, CH ₃ Leu), 0.90 (d, J = 6.0 Hz, 3H, CH ₃ Leu), 1.24 (t, J = 7.1 Hz, 3H, CH ₃ (Et)), 1.44 (s, 9H, (CH ₃ ) ₃ ), 1.61 (m, 3H, CH, CH ₂ Leu), 2.96 (m, 2H, CH ₂ Tyr), 3.31 (m, 2H, CH ₂ Tip), 3.86 (s, 3H, OCH ₃ ), 4.14 (m, 2H, CH ₂ (Et)), 4.30 (m, IH, CHa Tyr), 4.41 (m, IH, CHa Leu), 4.85 (m, 2H, CHa Trp and NHBoc), 6.40 (m, IH, NH), 6.60 (m, IH), 6.73 (d, J = 8.4 Hz, IH), 7.00 (t, J = 7.7 Hz, IH,

H5 Tip), 7.11 (m, 2H, aromatic H), 7.34 (d, J = 7.7 Hz, IH), 7.49 (d, J = 7.7 Hz, IH), 7.61 (d, J = 1.9 Hz, IH), 8.62 (broad s, IH, NH ind). ¹³ C NMR (75 MHz, CDCl ₃ ) δ 14.1 (CH ₃ Et), 22.2 (CH ₃ Leu), 22.9 (CH ₃ Leu), 25.6 (CH Leu), 27.7 (CH ₂ Trp), 28.3 ((CH ₃ ) ₃ ), 36.4 (CH ₂ Tyr), 41.4 (CH ₂ Leu), 51.7 (CHa Leu), 52.8 (CHa Trp), 55.5 (CHa Tyr), 56.3 (OCH ₃ ), 61.7 (OCH ₂ Et), 80.4 (C(CH ₃ ) ₃ ), 86.0 (C), 104.9 (C7 Trp), 110,8 (CH),110.9 (C), 117.8 (CH), 120.5 (CH), 124.3 (CH), 128.7 (C), 130.3 (CH), 130.8 (C), 134.7 (C), 140.1 (CH), 155.6 (C4 Tyr), 157.0 (CO Boc), 171.5, 171.6, 171.9 (2 CO amide, CO ester). Anal. Calcd. for C ₃₅ H ₄₄ N ₄ O ₇ BrI: C, 49.77; H, 5.47; N, 6.41. Found: C, 49.35; H ₅ 5.36; N, 6.77.

N-Boc-3-iodo-Tyr(Bn)-Aϊa-7-bromo-Trp-NHMe : compound A254. The peptide coupling was performed in CH ₂ Cl ₂ (4.4 mL) using HBr,7-bromo-Trp-NHMe (330 mg, 0.874 mmol), crude N-Boc-3-iodo-Tyr(Bn)-Ala-OH (521 mg, 0.918 mmol), EDC (124 mg, 0.96 mmol), HOBt (130 mg, 0.96 mmol) and NEt ₃ (370 μL, 2.62 mmol). The residue was subjected to flash chromatography on silica gel (5% MeOH/CH ₂ Cl ₂ ) to afford tripeptide N- Boc-3-iodo-Tyr(Bn)-Ala-7-bromo-Trp-NHMe (322 mg, 43%) as a white amorphous solid. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.18 (d, J = 7 Hz, 3H), 1.29 (s, 9H), 2.57 (d, J = 4 Hz, 3H), 2.81 (m, 2H), 3.11 (m, 2H), 4.07 (m, IH), 4.28 (m, IH), 4.43 (m, IH), 5.14 (s, 2H), 6.96 (m, 2H), 7.17-8.21 (m, 15H), 11.04 (s, IH). ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 25.6 (CH ₃ Ala), 27.7 (CH ₂ Trp), 28.0 (CH ₃ Boc and CH ₃ N), 35.7 (CH ₂ Tyr), 47.9 (CHa Ala), 53.2(CHa Trp), 55.6(CHa Tyr), 69.9 (CH ₂ Bn),78.0 (C Boc), 86.3 (CI), 104.0 (CBr) ₅ 111.6 (Cγ Trp), 112.5, 117.9, 119.6, 123.3, 124.9, 127.0, 127.6, 128.3, 128.9 (C), 130.4, 132.6 (C), 134.2 (C), 136.7 (C), 139.3 (C), 139.4, 155.1 (CO Boc), 171.2 (CONH), 171.4 (CONH), 171.6 (CONH). HRMS (ESI) calcd for C ₃₆ H ₄₁ BrIN ₅ O ₆ Na [(M+Na) ⁺ ] 868.1183, found 869.1175.

Ill) Preparation of macrocyclic peptides: compounds II

Macrocyclic peptides were prepared according to Berthelot, A.; Piguel, S.; Le Dour, G.; Vidal, J., Synthesis of macrocyclic peptide analogues of proteasome inhibitor TMC-95A. J. Org. Chem. 2003, 68, (25), 9835-9838.

A374F1 : described in the above article

A291 described in the above article

A389Flpl2 described in the above article

IV) Preparation of biaryl compounds: compounds III

General procedure for the preparation of biaryls as illustrated by the synthesis of:

Biaryl SP225F2:

N-(tert-butoxycarbonyl)-3-[(4,4,5,5-tetramethyl-l,3,2-dio xaboroIane-2-yl)]-Tyr(Bn)- AIa-OMe: A flame-dried Schlenk tube charged with Boc-4iodoTyr(Bn)-Ala-OMe (1.47 g, 2.53 mmol), KOAc (946 mg, 9.64 mmol), bis(pinacolato)diboron (773 mg, 3.04 mmol) and Pd(dρpf)Cl ₂ .CH ₂ Cl ₂ (166 mg, 8 mol %) was flushed with argon. Degassed DMSO (17 niL) was added and the reaction mixture was stirred at 80 ⁰ C for 16 h. The mixture was diluted with cold water, extracted with CH ₂ Cl ₂ and the combined organic extracts were washed with brine, dried over Na ₂ SO ₄ and the solvent was concentrated in vacuo. The brown oil was purified by flash chromatography on silica gel (20-40% AcOEt/Heptane) to give an inseparable mixture of the aryl boronate N-(tert-butoxycarbonyl)-3-[(4,4,5,5-tetramethyl- l,3,2-dioxaborolane-2-yl)]-Tyr(Bn)-Ala-OMe (57.5% yield estimated by ¹ H NMN) and the dipeptide Boc-Tyr(Bn)- AIa-OMe (939 mg, ca 7.8:1 ratio). Aryl boronate : ¹ H NMR (300 MHz, CDCl ₃ ) δ 1.37 (d, J = 7.5 Hz, 3H, CH ₃ Ala), 1.38 (s, 12H ₅ 4 CH ₃ boronic ester), 1.44 (s, 9H, (CH ₃ ) ₃ ), 2.99 (dd, J = 14 Hz, J = 6.7 Hz, IH, CH ₂ Tyr), 3.1 (dd, J = 14 Hz, J = 6.2 Hz, IH, CH ₂ Tyr), 3.72 (s, 3H, OCH ₃ ), 4.33 (m, IH, CHa Tyr), 4.53 (m, IH, CHa Ala), 4.97 (broad s, IH, NHBoc), 5.11 (s, 2H, CH ₂ (Bn)), 6.44 (m, IH, NH amide), 6.89 (d, J = 8,5 Hz, IH ₅ H-2), 7.26 (dd, J = 8.7 Hz, J = 2.2 Hz ₅ IH, H-3), 7.28-7.46 (m, 3 aromatic H), 7.53 (d, J = 2.2 Hz, IH, H5), 7.62 (m, 2 aromatic H). ¹³ C NMR (50 MHz, CDCl ₃ ) δ 18.4 (CH ₃ Ala), 24.9 (4 CH ₃ boronic ester), 28.3 ((CH ₃ ) ₃ ), 37.3 (CH ₂ Tyr), 48.2 (CHa Ala), 52.4 (OCH ₃ ), 55.7 (CHa Tyr), 70 (CH ₂ (Bn)), 80.2 (C (Boc)), 83.5 (2 C(CH ₃ ) ₂ ), 112.3 (CH(2)), 126.7-128.9 (8 aromatic C), 133.4 (C3), 137.5 (CH(5)), 155.3 (C(4)), 162.4 (CO (Boc)), 170.9, 172.8 (CO amide, CO ester). HRMS (LSMS with Cs ⁺ ) calcd for C ₃₁ H ₄₄ N ₂ O ₈ B [(M+H) ⁺ ] 583.3191, found 583.3184. A flask adapted with a condenser and charged with the unseparable mixture

of aryl boronate and dipeptide Boc-Tyr(Bn)-Ala-OMe (909 mg, 1.42 mmol based on the ratio 7.8 : 1 in favor of the aryl boronate), Z-7-bromoTrp-NHMe (488 mg, 1.13 mmol), P(o-tolyl) ₃ (86.4 mg, 20 mol %) and Pd(OAc) ₂ (32.2 mg, 10 mol %) was flushed with argon. Degassed dioxane (11 mL) and 1.4 mL of an aqueous solution OfNa ₂ CO ₃ (2.8 mmol, 2 M) were added. The resulting mixture was stirred at 85 °C for 3-4 h. The reaction mixture was passed through a pad of celite and the solvent was concentrated in vacuo. The greenish residue was purified by flash chromatography on silica gel (60-80% AcOEt/Heptane) and the biaryl compound was isolated as an amorphous pale yellow solid (547.5 mg, 60%). Only one atropoisomer was obtained. R _f 0.3 (80% AcOEt/Heptane). ¹ H NMR (500 MHz, CDCl ₃ ) δ 1.37 (s, 9H, (CH ₃ ) ₃ ), 1.38 (d, J = 7.5 Hz, 3H, CH ₃ Ala), 2.6 (d, J = 4.7 Hz , 3H, NHMe), 2.93 (dd, J = 13.9 Hz, J = 7.2 Hz, IH, CH ₂ Tyr), 3.15 (m, 2H, CH ₂ Tyr, CH ₂ Trp), 3.4 (dd, J = 13.9 Hz, J = 4.2 Hz, IH, CH ₂ Trp), 3.55 (s, 3H, OCH ₃ ), 4.35 (m, 3H, CHa Tyr, CHa Ala, CHa Trp), 4.99 (m, 2H, CH ₂ (Z)), 5.12 (s, 2H, CH ₂ (Bn)), 5.18 (broad s, IH, NHBoc), 5.65 (broad s, IH ₅ NHZ), 5.74 (broad s, IH, NHMe), 6.65 (m, IH, NH amide), 7.01-7.35 (m, 16 aromatic H), 7.65 (broad s, 1 aromatic H), 9.01 (broad s, IH, NH indole). ¹³ C NMR (75 MHz, CDCl ₃ ) δ 18.1 (CH ₃ Ala), 26.1 (NHCH ₃ ), 28.2 ((CH ₃ ) ₃ ), 28.8 (CH ₂ Trp), 38.3 (CH ₂ Tyr), 48 (CHa Ala), 52.4 (OCH ₃ ), 55.1, 55.8 (CHa Tyr, CHa Trp), 67, 70.8 (CH ₂ (Bn), CH ₂ (Z)), 80.2 ((CH ₃ ) ₃ C), 109-137 (25 aromatic C), 154.8, 155.5, 156 (C-OBn, 2 CO carbamate), 171.2, 172.1, 173.1 (2 CO amide, CO ester). HRMS (LSIMS with Cs ⁺ ) calcd for C ₄₅ H ₅₁ N ₅ O ₉ [M ⁺ ] 805.3687, found 805.3688.

Biaryl SP221:

N-(tert-butoxycarbonyl)-3-[(4,4,5,5-tetramethyl-l,3,2-dio xaborolane-2-yl)]-Tyr(Bn)-Leu- OMe: Same procedure as described above with Boc-4-iodo-Tyr(Bn)-Leu-OMe (2 g, 3.2 mmol), KOAc (1.09 g, 11.1 mmol), bis(pinacolato)diboron (986 mg, 3.88 mmol) and Pd(dppf)Cl ₂ .CH ₂ Cl ₂ (212 mg, 8 mol %) in DMSO (20 mL). The reaction mixture was stirred at 8O ⁰ C for 18 h followed by work-up. After purification by flash chromatography on silica gel (20-40% AcOEt/Heptane), an inseparable mixture of the aryl boronate (62% yield estimated by ¹ H RMN) and the dipeptide Boc-Tyr(Bn)-Leu-OMe was isolated as a white foam (ca 4.2 : 1 ratio). Aryl boronate : ¹ H NMR (300 MHz, CDCI ₃ ) δ 0.9 (d, J = 5.6 Hz, 3H, CH ₃ Leu), 0.92 (d, J = 5.6 Hz, 3H, CH ₃ Leu), 1.38 (s, 12H, 4 CH ₃ boronic ester), 1.43 (s, 9H,

(CHa) ₃ ), 1-44-1.6 (m, 3H, CH ₂ , CH Leu), 3.02 (m, 2H, CH ₂ Tyr), 3.7 (s, 3H, OCH ₃ ), 4.3 (m, IH, CHa Tyr), 4.56 (m, IH, CHa Leu), 4.9 (broad s, IH, NHBoc), 5.11 (s, 2H, CH ₂ (Bn)), 6.29 (m, IH, NH amide), 6.88 (d, J = 8.5 Hz ₅ IH, H-2), 7.27-7.42 (m, 4 aromatic H), 7.54 (d, J - 2.3 Hz, IH, H-5), 7.62 (m, 2 aromatic H). ¹³ C NMR (50 MHz, CDCl ₃ ) δ 21.9, 22.7 (CH ₃ Leu), 24.6 (CH Leu), 24.9 (4 CH ₃ boronic ester), 28.2 ((CH ₃ ) ₃ ), 37 (CH ₂ Tyr), 41.5 (CH ₂ Leu), 50.7 (CHa Leu), 52.2 (OCH ₃ ), 55.8 (CHa Tyr), 70 (CH ₂ (Bn)), 80.1 ((CH ₃ ) ₃ C), 83.5 (2 C(CH ₃ ) ₂ ), 112.3 (C-2), 126.7-136.9 (8 aromatic C), 133.3 (C-3), 137.5 (C-5), 155.4 (C-OBn), 162.4 (CO carbamate), 171.1, 172.7 (CO amide, CO ester). HRMS (ESI) calcd for C ₃₄ H ₄₉ N ₂ O ₈ BNa [(M+Na) ⁺ ] 647.3480, found 647.3489. Same procedure as described above with a mixture of N-(tert-butoxycarbonyl)-3-[(4,4,5,5- tetramethyl-l,3,2-dioxaborolane-2-yl)]-Tyr(Bn)-Leu-OMe and dipeptide Boc-Tyr(Bn)-Leu- OMe (100 mg, 0.135 mmol), Z-7-bromoTrp-OEt (48.1 mg, 0.11 mmol), P(o-tolyl)3 (8.3 mg, 20 mol %), Pd(OAc) ₂ (3 mg, 10 mol %) and 140 μL of an aqueous solution Of Na ₂ CO ₃ (0.28 mmol, 2 M) in degassed dioxane (1.1 niL). The reaction was stirred at 85 ⁰ C for 2 h. After purification by flash chromatography on silica gel (20-30% AcOEt/Heptane), the biaryl was isolated as a pale yellow solid (87.8 mg, 94%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 0.9 (m, 6H, 2 CH ₃ Leu), 1.2 (t, J = 7 Hz, 3H, CH ₃ ester), 1.36 (s, 9H, (CH ₃ ) ₃ ), 1.4-1.62 (m, 3H, CH ₂ CH Leu), 2.95 (dd, J = 13.5 Hz, J = 6.9 Hz, IH, CH ₂ Tyr), 3.15 (dd, J = 13.8 Hz, J = 6.4 Hz, IH, CH ₂ Tyr), 3.37 (m, 2H, CH ₂ Tip), 3.55 (s, 3H, OCH ₃ ), 4.13 (m, 2H, CH ₂ ester), 4.49 (m, 2H, CHa Tyr), 4.6 (m, IH, CHa Leu), 4.74 (m, IH, CHa Tip), 4.99 (m, 2H, CH ₂ (Z)), 5.13 (m, 3H, CH ₂ (Bn), NHBoc), 5.4 (broad d, J = 8.1 Hz, IH, NHZ), 6.24 (broad d, J = 8.2 Hz, IH, NH amide), 6.99-7.36 (m, 16 aromatic H), 7.54 (d, J = 7.8 Hz, 1 aromatic H), 9.09 (broad s, IH, NH indole). 13C NMR (50 MHz, CDCl ₃ ) δ 12.9 (CH ₃ ester), 20.7, 21.6, 23.6 (2 CH ₃ , CH Leu), 26.9 (CH ₂ Tip), 27.1 ((CH ₃ ) ₃ ), 37.1, 40.4 (CH ₂ Tyr, CH ₂ Leu), 49.6, 51.1, 53.5, 54 (CHa Leu, OCH ₃ , CHa Tyr, CHa Tip), 60.3 (CH ₂ ester), 65.7, 69.8 (CH ₂ (Bn), CH ₂ (Z)), 79.2 ((CH ₃ ) ₃ C), 108.3-135.8 (25 aromatic C), 153.7, 154.3, 154.7 (C-OBn, 2 CO carbamate), 170, 170.9, 171.9 (2 CO amide, CO ester). Anal. Calcd. for C ₄₉ H ₅₈ N ₄ O ₁₀ : C, 68.2; H, 6.77; N, 6.49. Found: C, 68.14; H, 6.82; N, 6.03.

847,99

Biaryl SP226F1 : Same procedure as described above with a mixture of aryl boronate N- (tert-butoxycarbonyl)-3-[(4,4,5,5-tetramethyl-l,3,2-dioxabor olane-2-yl)]-Tyr(Bn)-Leu-OMe and dipeptide Boc-Tyr(Bn)-Leu-OMe (4.2 : 1 ratio) (361 mg, 0.463 mmol), Z-7-bromoTrp- NHMe (191 mg, 0.44 mmol), P(o-tolyl) ₃ (15.1 mg, 10 mol %), Pd(OAc) ₂ (6.8 mg, 5 mol %) and 0.5 mL of an aqueous solution Of Na ₂ CO ₃ (1 mmol, 2 M) in degassed dioxane (3 mL). After purification by flash chromatography on silica gel (40-60% AcOEt/Heptane), the biaryl was isolated as a pale yellow solid (269 mg, 72%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 0.91 (m, 6H, 2 CH ₃ Leu), 1.38 (s, 9H, (CH ₃ ) ₃ ), 1.46-1.64 (m, 3H, CH ₂ CH Leu), 2.61 (d, J = 4.7 Hz , 3H, NHMe), 2.96 (dd, J = 14 Hz, J = 6.4 Hz, IH, CH ₂ Tyr or Trp), 3.15 (dd, J = 14 Hz, J = 7.5 Hz, 2H, CH ₂ Tyr, Trp), 3.43 (dd, J = 14 Hz, J = 3.6 Hz, IH, CH ₂ Trp or Tyr), 3.51 (s, 3H, OCH ₃ ), 4.48 (m, 2H, CHa Tyr, CHa Trp), 4.6 (m, 3H, CHa Leu), 4.99 (s, 2H, CH ₂ (Z)), 5.13 (s, 3H, CH ₂ (Bn), NHBoc), 5.64 (m, 2H, NHZ, NHMe), 6.29 (broad d, J = 8.4 Hz, IH, NH amide), 7.01-7.35 (m, 16 aromatic H), 7.65 (m, 1 aromatic H), 9.07 (broad s, IH, NH indole). ¹³ C NMR (50 MHz, CDCl ₃ ) δ 22.1, 23.1 (CH ₃ Leu), 25.1 (CH Leu), 26.6 (NHCH ₃ ), 28.6 ((CHa) ₃ ), 29.3 (CH ₂ Trp), 38.6 (CH ₂ Tyr), 41.7 (CH ₂ Leu), 51.1 (CHa Leu), 52.6 (OCH ₃ ), 55.9, 56.2 (CHa Tyr, CHa Trp), 67.4, 71.2 (CH ₂ (Bn), CH ₂ (Z)), 80.2 ((CHs) ₃ C), 110.1- 137.8 (25 aromatic C), 155.2, 155.9, 156.1 (C-OBn, 2 CO carbamate), 171.8, 172.5, 173.5 (2 CO amide, CO ester). HRMS (ESI) calcd for C ₄₈ H ₅₇ N ₅ O ₉ Na [(M+Na) ⁺ ] 870.4054, found 870.4068.

V) Preparation of non halogenated tripeptides: compounds IV-2

General procedure for the preparation of tripeptides as illustrated by the synthesis of

N-Boc-Tyr(Bn)-Ala-Trp-NHPh : compound SP303r2. To a solution of Trp-NHPh (202 mg, 0.723 mmol) in CH ₂ Cl ₂ ZDMF (3 mL, 1/1) at 0 ⁰ C were successively added N-Boc- Tyr(Bn)-Ala-OH (320 mg, 0.723 mmol), EDC (153.6 mg, 0.8 mmol) and HOBt (108 mg, 0.8 mmol). The resulting mixture was allowed to warm up to room temperature overnight. The solvent was evaporated and the crude was triturated with water. After filtration, the solid was collected and purified by precipitation in CH ₂ Cl ₂ ZMeOH to give a white amorphous solid

(160.3 mg, 31%). ¹ H NMR (300 MHz, DMSOd ₆ ) δ 1.22 (d, J = 7 Hz, 3H, CH ₃ ), 1.28 (s, 9H, (CH ₃ ) ₃ ), 2.63 (m, IH, CH ₂ ), 2.89 (m, IH, CH ₂ ), 3.06 (dd, J = 14.5 Hz, 7.5 Hz, IH, CH ₂ ), 3.2 (dd, J = 14.5 Hz, 6.1 Hz, IH, CH ₂ ), 4.1 (m, IH, CHa), 4.34 (m, IH, CHa), 4.69 (m, IH ₃ CHa), 5.02 (s, 2H, CH ₂ (Bn)), 6.88 (d, J = 8.5 Hz, 2 aromatic H Tyr), 6.94 (m, 2 aromatic H Trp), 7.03 (t, J = 7.4 Hz, 2 aromatic H Trp), 7.16 (m, 3 aromatic H), 7.25-7.42 (m, 8H), 7.58 (m, 3H), 8.02 (d, J = 7.2 Hz, IH), 8.18 (d, J = 7.6 Hz, IH), 10 (s, IH, NH), 10.8 (s, IH, NH). ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 18.2 (CH ₃ ), 27.7 (CH ₂ Trp), 28.1 ((CH ₃ ) ₃ ), 36.3 (CH ₂ Tyr), 48.1, 54.2, 55.7 (CHa Ala, Tyr, Trp), 69 (CH ₂ (Bn)), 78 (C(CH ₃ ) ₃ ), 109.5, 111.2, 114.2, 118.2, 118.4, 119.4, 120.8, 123.3, 123.5, 127.3, 127.5, 127.7, 128.3, 128.6, 130.1, 130.3, 135.9, 137.2, 138.8 (25 aromatic C), 155.2, 156.8 (Car-O, CO carbamate), 170, 171.5, 172.1 (3 CO amide). Anal. Calcd. for C ₄₁ H ₄₅ N ₅ O ₆ , 0.5 H ₂ O: C, 69.08; H, 6.5; N, 9.82. Found: C, 68.82; H, 6.34; N, 9.79.

N-Boc-Tyr(Bn)-Leu-Trp-NHPh : compound A424P. Same procedure as above with Trp- NHPh (67.6 mg, 0.242 mmol), N-Boc-Tyr(Bn)-Leu-OH (117 mg, 0.242 mmol), EDC (49 mg, 0.25 mmol) and HOBt (35 mg, 0.25 mmol) in CH ₂ C1 ₂ /DMF (3.6 mL, 1/1). The crude residue was dissolved in ether and precipitated with heptane to give a beige solid (135.6 mg, 75 %). ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 0.83 (d, 3H, J = 6.4 Hz, CH ₃ ), 0.87 (d, 3H, J = 6.4 Hz, CH ₃ ), 1.22-1.44 (m, 2H, CH ₂ Leu), 1.30 (s, 9H, (CH ₃ ) ₃ ), 1.44 (m, IH, CH Leu), 2.46-2.89 (m, 2H, CH ₂ ), 3.13 (m, 2H, CH ₂ ), 4.12 (m, IH, CHa), 4.38 (m, IH, CHa), 4.70 (m, IH, CHa), 5.02 (s, 2H, CH ₂ (Bn)), 6.87-7.58 (m, 2OH, 19 aromatic H and NH Boc), 7.94 (d, J = 8.2 Hz, IH), 8.17 (d, J = 7.5 Hz, IH), 10.0 (s, IH, NH), 10.83 (s, IH, NH). ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 21.5, 23.1, 23.9 (CH-(CH ₃ ) ₂ ), 27.7 (CH ₂ Trp), 28.1 ((CH ₃ ) ₃ ), 36.2, 40.9 (CH ₂ Tyr, CH ₂ Leu), 50.9, 54.1, 55.7 (CHa Tyr, Leu, Trp), 69.0 (CH ₂ (Bn)), 78.0 (C(CH ₃ ) ₃ ), 109.6, 111.2, 114.2, 118.2, 118.4, 119.3, 120.8, 123.3, 123.4, 127.3, 127.6, 127.7, 128.3, 128,6, 130.1, 130.3, 135.9, 137.2, 138.8 (25 aromatic C) ₅ 155.2, 156.8 (Car-O, CO carbamate), 170, 171.6,, 171.9 (3 CO amide). Anal. Calcd. for C ₄₄ H ₅₁ N ₅ O ₆ : C, 70.85; H, 6.89; N, 9.39. Found: C ₅ 70.52; H, 7.10; N, 9.33.

N-Boc-Tyr(Bn)-Asn-Trp-NHPh : compound SP314C2. Same procedure as above with Trp-NHPh (102.9 mg, 0.368 mmol), N-Boc-Tyr(Bn)-Asn-OH (178.9 mg, 0.368 mmol), EDC (78.2 mg, 0.41 mmol) and HOBt (55 mg, 0.41 mmol) in CH ₂ Cl ₂ ZDMF (1.5 niL, 1/1). The crude residue was purified by flash column chromatography on silica gel (0-5% MeOH/CH ₂ Cl ₂ ) to give an off-white solid (82.8 mg, 30 %). ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.28 (S ₅ 9H, (CH ₃ ) ₃ ), 2.57-3.26 (m, 6H, CH ₂ Tyr, CH ₂ Trp, CH ₂ Asn), 4.11 (m, IH, CHa), 4.58 (m, 2H, 2 CHa), 5.03 (broad s, 2H, CH ₂ (Bn)), 6.86-7.65 (m, 22H), 8.2 (m, 2H), 9.82 (s, IH), 10.78 (s, IH) . ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 27.3 (CH ₂ Trp), 28.1 ((CH ₃ ) ₃ ), 36.4, 37 (CH ₂ Tyr, CH ₂ Asn), 49.5, 54.5, 55.7 (CHa Tyr, Asn, Trp), 69 (CH ₂ (Bn)), 78.1 (C(CH ₃ ) ₃ ), 109.7, 110, 111.2, 114.2, 118.2, 119.6, 120.8, 123.4, 123.6, 127.2, 127.5, 127.7, 128.3, 128.5, 130.1, 136, 137.2, 138.7 (25 aromatic C), 155.2, 156.8 (Car-O, CO carbamate), 170, 171, 171.7, 171.9 (4 CO amide). Anal. Calcd. for C ₄₂ H ₄₆ N ₆ O ₇ : C, 67.54; H, 6.21; N, 11.25. Found: C, 67.14; H, 6.27; N, 11.27.

N-Boc-Tyr(Bn)-Arg(Z) ₂ -Trp-NHPh : compound SP310C. Same procedure as above with Trp-NHPh (100 mg, 0.358 mmol), N-Boc-Tyr(Bn)-Arg(Z) ₂ -OH (286 mg, 0.359 mmol), EDC (75.8 mg, 0.395 mmol) and HOBt (53.4 mg, 0.395 mmol) in CH ₂ C1 ₂ /DMF (1.8 mL, 1/1). The crude residue was purified by flash column chromatography on silica gel (0-1% MeOHZCH ₂ Cl ₂ ) to give a yellow solid (157.4 mg, 41%). ¹ H NMR (300 MHz, DMSOd ₆ ) δ 1.25 (s, 9H, (CH ₃ ) ₃ ), 1.63 (m, 4H, 2 CH ₂ Arg), 2.63 (m, IH, CH ₂ Tyr), 2.86 (m, IH, CH ₂ Tyr), 3.04 (dd, J = 14.6 Hz, J = 7.5 Hz, IH, CH ₂ Trp), 3.18 (dd, J = 14.5 Hz, J = 6.2 Hz, IH, CH ₂ Trp), 3.85 (m, 2H, CH ₂ Arg), 4.11 (m, IH, CHa), 4.36 (m, IH, CHa), 4.7 (m, IH, CHa), 5.01 (s, 4H, CH ₂ (Bn), CH ₂ (Z)), 5.17 (m, 2H, CH ₂ (Z)), 6.85-7.54 (m, 30H), 7.94 (d, J = 8 Hz,

IH) ₃ 8.24 (d, J = 7.5 Hz, IH), 9.15 (broad s, 2H), 10 (s, IH), 10.8 (s, IH) .. ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 24.9, 29.7 (2 CH ₂ Arg), 27.8 (CH ₂ Trp), 28 ((CH ₃ ) ₃ ), 36.3 (CH ₂ Tyr), 44.3 (CH ₂ Arg), 52.2, 54.2, 55.8 (CHa Tyr, Arg ₅ Trp), 66.1, 68.1, 69.1 (CH ₂ (Bn), 2 CH ₂ (Z)), 78 (C(CH ₃ ) ₃ ), 109.5, 111.2, 114.2, 118.2, 118.4, H9.4, 120.8, 123.2, 123.5, 127.3, 127.5, 127.6, 127.7, 127.8, 128.1, 128.2, 128.3, 128.4, 128.6, 130.1, 130.2, 135.2, 136, 137, 137.2, 138.8 (37 aromatic C), 155, 155.2, 156.8, 159.6, 162.9 (Car-O, 3 CO carbamate, imine), 170, 171.3, 171.6 (3 CO amide). Anal. Calcd. for C ₆₀ H ₆₄ N ₈ O ₁₀ , 1 H ₂ O: C, 67.02; H, 6.18; N, 10.42. Found: C, 67.34; H, 6.05; N, 10.27.

N-Boc-Tyr(Bn)-Lys(Boc)-Trp-NHPh : compound SP306P. Same procedure as above with Trp-NHPh (99.6 mg, 0.356 mmol), N-Boc-Tyr(Bn)-Lys(Boc)-OH (214 mg, 0.356 mmol), EDC (75.3 mg, 0.392 mmol) and HOBt (53.7 mg, 0.397 mmol) in CH ₂ Cl ₂ ZDMF (1.5 mL, 1/1). The crude residue was triturated with MeOH/pentane to afford a white solid (193.5 mg, 63%). ¹ H NMR (300 MHz, DMSOd ₆ ) δ 1.21-1.62 (m, 6H, 3 CH ₂ Lys), 1.3 (s, 9H, (CHs) ₃ ), 1.36 (s, 9H ₅ (CH ₃ ) ₃ ), 2.64 (m, IH ₅ CH ₂ Tyr), 2.87 (m, 3H, CH ₂ Lys, CH ₂ Tyr), 3.05 (dd, J = 14.7 Hz, J = 7.7 Hz, IH ₅ CH ₂ Trp), 3.19 (dd, J = 14.7 Hz, J = 6.2 Hz, IH ₅ CH ₂ Trp), 4.11 (m, IH ₅ CHa) ₅ 4.31 (m, IH, CHa) ₅ 4.71 (m ₅ IH ₅ CHa) ₅ 5.02 (s, 2H ₅ CH ₂ (Bn)), 6.71 (m, IH), 6.87-7.42 (m, 17H), 7.58 (m, 3H), 7.9 (d, J = 7.3 Hz, IH) ₅ 8.18 (d, J = 7.2 Hz, IH) ₅ 10 (s, IH), 10.8 (s, IH) . ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 22.5, 29.2, 32 (3 CH ₂ Lys), 27.8 (CH ₂ Trp), 28.1, 28.2 (2 (CH ₃ ) ₃ ), 36.3 (CH ₂ Tyr), 39.8 (CH ₂ Lys), 52.4, 54.2, 55.8 (CHa Tyr, Lys, Trp), 69.1 (CH ₂ (Bn)), 77.3, 78.1 (2 C(CH ₃ ) ₃ ), 109.6, 111.2, 114.3, 118.2, 118.4, 119.4, 120.8, 123.3, 123.4, 127.3, 127.5, 127.7, 128.3, 128.6, 130.1, 130.2, 136, 137.2, 138.8 (25 aromatic C) ₅ 155.2, 155.5, 156.8 (Car-O, 2 CO carbamate), 170.1, 171.5, 171.6 (3 CO amide). Anal. Calcd. for C ₄₉ H ₆₀ N ₆ O ₈ , 1.5 H ₂ O: C, 66.27; H, 7.15; N, 9.46. Found: C, 66.42; H, 6.96; N, 9.34.

N-Boc-Tyr(Bn)-Ala-Trp-NHCH ₂ Ph : compound SP304R. Same procedure as above with Trp-NHCH ₂ Ph (194.5 nig, 0.663 mmol), N-Boc-Tyr(Bn)-Ala-OH (293.5 mg, 0.663 mmol), EDC (141.5 mg, 0.73 mmol) and HOBt (99 mg, 0.73 mmol) in CH ₂ C1 ₂ /DMF (2.8 mL, 1/1). The crude residue was triturated with Et ₂ θ/pentane to afford a white solid (215.5 mg, 45%). ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.22 (d, J = 6.5 Hz, 3H, CH ₃ ), 1.3 (s, 9H, (CH ₃ ) ₃ ), 2.61 (m, IH, CH ₂ ), 2.87 (m, IH, CH ₂ ), 3.02 (dd, J = 14.1 Hz, 7 Hz, IH, CH ₂ ), 3.17 (dd, J = 14.1 Hz, 6.1 Hz, IH, CH ₂ ), 4.24 (m, 4H, CH ₂ (Bn) Trp, 2 CHa), 4.58 (m, IH, CHa), 5.04 (s, 2H, CH ₂ (Bn)), 6.89-7.44 (m, 19 aromatic H), 7.61 (d, J = 7.7 Hz, IH) ₅ 8.04 (d, J = 7.1 Hz, IH), 8.1 (d, J = 7.6 Hz, IH), 8.42 (m, IH), 10.8 (s, INH). ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 18.2 (CH ₃ ), 27.8 (CH ₂ Trp), 28.1 ((CH ₃ ) ₃ ), 36.3 (CH ₂ Tyr), 42 (CH ₂ (Bn (Trp)), 48.1, 53.5, 55.7 (CHa Ala, Tyr, Tip), 69 (CH ₂ (Bn)), 78 (C(CH ₃ ) ₃ ), 109.7, 111.2, 114.2, 118.2, 118.4, 120.8, 123.6, 126.5, 126.9, 127.3, 127.5, 127.7, 128.1, 128.3, 130.2, 130.3, 136, 137.2, 139 (25 aromatic C), 155.2, 156.8 (Car-O, CO carbamate), 171, 171.4, 171.8 (3 CO amide). Anal. Calcd. for C ₄₂ H ₄₇ N ₅ O ₆ : C, 70.27; H, 6.6; N, 9.76. Found: C, 69.97; H, 6.73; N, 9.65.

N-Boc-Tyr(Bn)-Leu-Trp-NHCH ₂ Ph : compound A414P. Same procedure as above with Trp-NHCH ₂ Ph (136.8 mg, 0.467 mmol), N-Boc-Tyr(Bn)-Leu-OH (227 mg, 0.467 mmol), EDC (94 mg, 0.49 mmol) and HOBt (66 mg, 0.49 mmol) in CH ₂ Cl ₂ (5 mL). The reaction mixture was diluted by CH ₂ Cl ₂ , washed with 2M aqueous Na ₂ CO ₃ , then 5% aqueous KHSO ₄ and water. After drying of the organic phase over Na ₂ SO ₄ and concentration in vacuo, the product was afforded as a white solid (232.9 mg, 66%). ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 0.82 (d, 3H, J = 6.4 Hz, CH ₃ ), 0.86 (d, 3H, J = 6.4 Hz, CH ₃ ), 1.30 (s, 9H, (CH ₃ ) ₃ ), 1.45 (m,

2H, CH ₂ Leu),1.59 (m, IH, CH Leu), 2.59-2.88 (m, 2H, CH ₂ ), 3.07 (m, 2H, CH ₂ ), 4.08 (m, IH, CHa), 4.20(d, 2H, J - 6 Hz, NCH ₂ Ph) ₅ 4.36 (m, IH, CHa) ₅ 4.57 (m, IH, CHa), 5.02 (s, 2H, OCH ₂ (Bn)), 6.87-7.4 (m, 19H, 18 aromatic H and NH Boc), 7.58 (d, J = 7.7 Hz, 1H),7.95 (d, J = 8 Hz, IH), 8,11 (d, J = 7.9 Hz ₅ IH) ₅ 8.4 (t, IH, J = 6 Hz ₅ NH Bn), 10.84 (s, IH, NH). ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 21.5, 23.1 (CH ₃ Leu), 23.9 (CH ₂ Leu), 27.8 (CH ₂ Tip), 28.1 ((CHs) ₃ ), 41 (CH ₂ Tyr), 42 (CH ₂ (Bn (Tip)), 51, 53.5, 55.8 (CHa Leu, Tyr, Tip), 69 (CH ₂ (Bn)), 78 (C(CH ₃ ) ₃ ), 109.7, 111.2, 114.2, 118.2, 118.4, 120.8, 123.5, 126.5, 126.9, 127.3, 127.5, 127.7, 128.0, 128.3, 130.2, 130.3, 136, 137.2, 139 (25 aromatic C), 155.2, 156.8 (Car-O, CO carbamate), 171, 171.6, 171.7 (3 CO amide). Anal. Calcd. for C ₄₅ H ₅₃ N ₅ O ₆ : C, 71.12; H, 7.03; N, 9.22. Found: C, 70.85; H, 9.96; N, 9.07.

N-Boc-Tyr(Bn)-Arg(Z) ₂ -Trp-NHCH ₂ Ph : compound SP31SC2. Same procedure as above with Trp-NHCH ₂ Ph (139 mg, 0.474 mmol), N-Boc-Tyr(Bn)-Arg(Z) ₂ -OH (378 mg, 0.474 mmol), EDC (100.2 mg, 0.523 mmol) and HOBt (70.8 mg, 0.523 mmol) in CH ₂ C1 ₂ /DMF (3 mL, 1/1). The crude residue was triturated with MeOH/pentane and the resulting white solid was purified by flash column chromatography on silica gel (0-1% MeOH/CH ₂ Cl ₂ ) to give a white solid (249 mg, 49%). ¹ H NMR (300 MHz, DMSOd ₆ ) δ 1.25 (s, 9H, (CHa) ₃ ), 1-54 (m, 4H, 2 CH ₂ Arg), 2.62-3.1 (m, 4H, CH ₂ Tyr, CH ₂ Trp), 3.85 (m, 2H, CH ₂ Arg), 4.17 (m, 3H, CH ₂ Bn (Trp), CHa), 4.33 (m, IH, CHa), 4.58 (m, IH, CHa), 5.02 (s, 4H, CH ₂ (Bn), CH ₂ (Z)), 5.21 (m, 2H ₅ CH ₂ (Z)), 6.81-7.39 (m, 29H), 7.58 (d, J = 7.2 Hz ₅ IH) ₅ 7.97 (m, IH) ₅ 8.12 (m, IH), 8.37 (m, IH) ₅ 9.16 (broad s, 2H) ₅ 10.8 (s, IH) . ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 24.8, 29.6 (2 CH ₂ Arg), 27.8 (CH ₂ Trp), 28 ((CH ₃ ) ₃ ), 36.3 (CH ₂ Tyr), 42 (CH ₂ (Bn) Trp), 44.3 (CH ₂ Arg), 52.2, 53.7, 55.9 (CHa Tyr, Arg, Trp), 66.1, 68.1, 69 (CH ₂ (Bn) ₅ 2 CH ₂ (Z)), 78 (C(CHs) ₃ ), 109.6, 111.2, 114.2, 118.2, 118.4, 120.8, 123.6, 126.5, 126.9, 127.3, 127.5, 127.7, 127.8, 127.9, 128, 128.2, 128.3, 128.4, 128.5, 130.1, 135.2, 136, 137, 137.1, 138.9 (37 aromatic C), 155, 155.2, 156.8, 159.6, 162.9 (Car-O, 3 CO carbamate, imine), 171, 171.1, 171.6 (3 CO amide). Anal. Calcd. for C ₆₁ H ₆₆ N ₈ O ₁₀ , 0.5 H ₂ O: C, 67.82; H, 6.25; N, 10.37. Found: C ₅ 67.69; H, 6.13; N, 10.29.

N-Boc-Tyr(Bn)-Lys(Boc)-Trp-NHCH ₂ Ph : compound SP307P. Same procedure as above with Trp-NHCH ₂ Ph (120.7 mg, 0.411 mmol), N-Boc-Tyr(Bn)-Lys(Boc)-OH (246.9 mg, 0.411 mmol), EDC (87.1 mg, 0.45 mmol) and HOBt (62.1 mg, 0.46 mmol) in CH ₂ C1 ₂ /DMF (1.8 mL, 1/1). The crude residue was triturated with MeOH/pentane to afford a white solid (243 mg, 67%). ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.2-1.61 (m, 6H, 3 CH ₂ Lys), 1.29 (s, 9H, (CHs) ₃ ), 1-36 (s, 9H, (CH ₃ ) ₃ ), 2.64 (m, IH, CH ₂ Tyr), 2.86 (m, 3H, CH ₂ Lys, CH ₂ Tyr), 3 (dd, J = 14.1 Hz, J = 7 Hz, IH, CH ₂ Trp), 3.14 (dd, J = 14.1 Hz, J = 6 Hz, IH, CH ₂ Trp), 4.11 (m, IH, CHa), 4.26 (m, 3H, CH ₂ (Bn Trp), CHa), 4.59 (m, IH, CHa), 5.02 (broad. s, 2H, CH ₂ (Bn)), 6.72 (m, IH), 6.85-7.4 (m, 19H), 7.59 (d, J = 7.6 Hz, IH), 7.89 (d, J = 7.4 Hz, IH), 8.1 (d, J - 7.4 Hz, IH), 8.38 (m, IH), 10.8 (s, IH) . ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 22.4, 29.2, 32 (3 CH ₂ Lys), 27.8 (CH ₂ Trp), 28.1, 28.2 (2 (CH ₃ ) ₃ ), 36.3 (CH ₂ Tyr), 39.8 (CH ₂ Lys), 42 (CH ₂ (Bn) Trp), 52.4, 53.5, 55.8 (CHa Tyr, Lys, Trp), 69.1 (CH ₂ (Bn)), 77.3, 78 (2 C(CH ₃ ) ₃ ), 109.7, 111.2, 114.3, 118.2, 118.4, 120.8, 123.5, 126.5, 126.9, 127.3, 127.5, 127.7, 128, 128.3, 130.1, 130.2, 136, 137.2, 139 (25 aromatic C), 155.2, 155.5, 156.8 (Car-0, 2 CO carbamate), 171, 171.3, 171.6 (3 CO amide). Anal. Calcd. for C ₅₀ H ₆₂ N ₆ O ₈ , 2 H ₂ O: C, 65.91; H, 7.30; N, 9.22. Found: C, 65.59; H, 7.06; N, 9.09.

N-Boc-Tyr(Bn)-Asn-Trp-NHCH2Ph : compound A416. Same procedure as above with Trp-NHCH ₂ Ph (63.03 mg, 0.215 mmol), N-Boc-Tyr(Bn)-Asn-OH (105.0 mg, 0.216 mmol), EDC (44 mg, 0.226 mmol) and HOBt (31 mg, 0.226 mmol) in CH ₂ Cl ₂ ZDMF (1.5 mL/1.5 mL). After treatment, the crude residue was triturated with CH ₂ Cl ₂ /pentane to afford a white solid (78 mg, 48%). ¹ H NMR (300 MHz, DMSO-d ₆ ) δ ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.29 (s, 9H, (CH ₃ ) ₃ ), 2.30-3.26 (m, 6H, CH ₂ Tyr, CH ₂ Trp, CH ₂ Asn), 4.08 (m, IH, CHa),

4.24 (s, 2H, NCH ₂ Ph), 4.49 (m, IH, CHa) ₅ 4.56 (m, IH, CHa), 5.03 (broad s, 2H, OCH ₂ PIi), 6.88-7.56 (m, 17H), 8.17. (d, J = 8 Hz, IH, NH), 8.23 (d, J = 8 Hz, IH ₃ NH), 8.51 (t, J = 6 Hz, IH, NH), 10.84 (s, IH, NH) . ¹³ C NMR (75 MHz, CD ₃ OD) δ 28.5 (CH ₂ Tip), 28.6 ((CH ₃ ) ₃ ), 37.3, 38.1 (CH ₂ Tyr, CH ₂ Asn), 44.1 (NCH ₂ Ph),51.7, 55.9, 57.6 (CHa Tyr, Asn, Trp),71.0 (OCH ₂ Ph), 80.9 (C Boc), 110.9 (C), 112.4, 115.9, 116.0, 119.4, 119.9, 122.5, 124.8, 128.0, 128.5, 128.6, 128.8, 129.4, 129.5, 130.6 (C), 131.4 (CH), 138.0, 138.8, 139.5 (aromatic C), 157.9, 159.1 (aromatic C and CO Boc), 172.7,173.6, 174.4, 174.9 (CO amide).

N-Boc-Tyr(Bn)-Gly-Trp-NHCH ₂ Ph : compound PSVIlR. Same procedure as above with Trp-NHCH ₂ Ph (82.43 mg, 0.281 mmol), N-Boc-Tyr(Bn)-Gly-OH (109.26 mg, 0.255 mmol), EDC (53.87 mg, 0.281 mmol) and HOBt (37.97 mg, 0.281 mmol) in CH ₂ C1 ₂ /DMF (2 mL/0.8 mL). The crude residue was triturated with CH ₂ Cl ₂ /ρentane to afford a white solid

(113.76, 63%). mp 183 °C. ¹ H NMR (300 MHz, CDCI ₃ ) δ 1.38 (s, 9H, (CH ₃ ) ₃ ), 2.81 (m,

2H, CH ₂ Tyr), 3.26 (m, 2H), 3.72 (m, 2H, CH ₂ GIy), 4.30 (m, 3H, CHa and CH ₂ Bn), 4.80 (m, IH, CHα),5.0 (broad s, 3H, NHBoc and CH ₂ O), 6.72 (m, IH, NH GIy), 6.90-7.40 (m, 20 H, aromatic H and NH), 7.63 (d, IH, J = 7 Hz), 8.35 (s, IH, NH indole). ¹³ C NMR (75 MHz,

CDCl ₃ ) δ 28.3 (CH ₂ Trp and (CH ₃ ) ₃ ), 37.5 (CH ₂ Tyr), 43.1, 43.5 (CH ₂ (Bn), CH ₂ GIy), 54.0,

55.8 (CHa Tyr, Trp), 70.0 (CH ₂ (Bn)), 80.4 (C(CEk) ₃ ), 110.2, 111.3, 114.9, 118.7, 119.5,

122.0, 123.4, 127.2, 127.4, 127.5, 127.7, 128.0, 128.5, 128.6, 128.7, 130.3, 136.1, 137.0, 137.9 (20 aromatic C), 155.7, 157.7 (Car-O, CO carbamate), 168.9, 171.4, 172.5 (3 CO amide). Anal. Calcd. for C ₄₁ H ₄₅ N ₅ O ₆ , 1 H ₂ O: C, 68.22; H, 6.56; N, 9.71. Found: C, 68.50; H,

6.48; N, 9.98.

N-Boc-Tyr(Bn)~Ala-Trp-NH(4-OH)Ph : compound SP313P. Same procedure as above with Trp-NHPhOH (75 mg, 0.254 mmol), N-Boc-Tyr(Bn)-Ala-OH (106.4 mg, 0.24 mmol), EDC (53.8 mg, 0.28 mmol) and HOBt (37.9 mg, 0.28 mmol) in CH ₂ Cl ₂ ZDMF (1.5 mL, 1/1). The crude residue was triturated with Et ₂ O to afford a pale brown solid (72.8 mg, 42%). ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.21 (d, J = 6.7 Hz, 3H, CH ₃ ), 1.28 (s, 9H ₅ (CH ₃ ) ₃ ), 2.62-3.21 (m, 4H, CH ₂ Tyr, CH ₂ Trp), 4.1 (m, IH, CHa), 4.32 (m, IH, CHa), 4.64 (m, IH, CHa), 5.02 (s, 2H, CH ₂ (Bn)), 6.66 (d, J = 8.4 Hz, IH), 6.87-7.39 (m, 17H), 7.6 (d, J = 7 Hz ₅ IH) ₅ 8 (d, J - 6.9 Hz, IH), 8.1 (d, J = 7.1 Hz ₅ IH) ₅ 9.18 (s, 1 NE) ₅ 9.74 (s, 1 NH) ₅ 10.8 (s, IH, NH). ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 18.2 (CH ₃ ), 27.8 (CH ₂ Trp), 28 ((CH ₃ ) ₃ ), 36.2 (CH ₂ Tyr), 48.1, 53.9, 55.7 (CHa Ala, Tyr, Trp), 69 (CH ₂ (Bn)), 78 (C(CH ₃ ) ₃ ), 109.6, 111.I ₅ 114.2, 114.9, 118.I ₅ 118.4, 120.7, 121, 123.4, 127.2, 127.5, 127.6, 128.3, 130.1, 130.2, 130.3, 135.9, 137.1 (24 aromatic C), 153.3, 155.2, 156.7 (2 Car-O, CO carbamate), 169.2, 171.4, 171.9 (3 CO amide). Anal. Calcd. for C ₄₁ H ₄₅ N ₅ O ₇ , 2 H ₂ O: C, 65.14; H, 6.53; N, 9.26. Found: C, 65.08; H, 6.29; N, 9.92. HRMS (ESI) calcd for C ₄₁ H ₄₅ N ₅ O ₇ Na [(M+Na) ⁺ ] 742.3217, found 742.3223.

N-Boc-Tyr(Bn)-Leu-Trp-NH(4-OH)Ph : compound A418P. Same procedure as above with Trp-NH(4-OH)Ph (58.99 mg, 0.2 mmol), N-Boc-Tyr(Bn)-Leu-OH (97 mg, 0.2 mmol), EDC (41 mg, 0.21 mmol) and HOBt (29 mg, 0.21 mmol) in CH ₂ Cl ₂ ZDMF (3 mL IZl). The reaction mixture was diluted by CH ₂ Cl ₂ , washed with 2M aqueous Na ₂ CO ₃ , then 5% aqueous KHSO ₄ and water. After drying of the organic phase over Na ₂ SO ₄ and concentration in vacuo, the crude residue was triturated with Et ₂ O to afford a beige solid (118.36 mg, 78%). ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 0.83 (d, 3H, J = 6.4 Hz, CH ₃ ), 0.87 (d, 3H, J = 6.5 Hz, CH ₃ ), 1.30 (s, 9H, (CH ₃ ) ₃ ), 1.43 (m, 2H, CH ₂ Leu), 1.6 (m, IH, CH Leu), 2.64-2.89 (m, 2H, CH ₂ ), 3.10 (m, 2H, CH ₂ ), 4.12 (m, IH, CHa), 4.37 (m, IH, CHa), 4.65 (m, IH, CHa), 5.02 (s, 2H, OCH ₂ (Bn)), 6.65-7.43 (m, 18H ₅ 17 aromatic H and NH Boc), 7.59 (d, J = 7.7 Hz, IH), 7.94 (d, J =F 8.9 Hz, IH), 8.10 (d, J = 7.6 Hz, IH), 9.18 (s, IH, OH), 9.72 (s, IH ₅ NHAr), 10.81 (s, IH, NH). ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 22.0, 23.4 (CH ₃ Leu), 25.7 (CH Leu), 28.7 ((CH ₃ ) ₃ ), 29 (CH ₂ Trp), 38.1, 41.7 (CH ₂ Leu, Tyr), 53.5, 56.1, 57.2 (CHa Leu, Tyr, Trp), 71 (CH ₂ (OBn)), 80.7 (C(CH ₃ ) ₃ ), 110.8, 112.3, 115.9, 116.1, 119.5, 119.9, 122.5, 123.8, 124.6,

128.5, 128.8, 128.9, 129.5, 130.7, 131.0, 131.4, 138.0, 138.8 (24 aromatic C), 155.6, 157.7, 159.1 (2 Car-O, CO carbamate), 171.7, 174.3, 174.7 (3 CO amide). Anal. Calcd. for C ₄₄ H _5I N ₅ O ₇ , 0.5 H ₂ O: C, 68.55; H, 6.80; N, 9.08. Found: C ₅ 68.21; H, 6.68; N, 9.09.

N-Boc-Tyr(Bn)-Asn-Trρ-NH(4-OH)Ph : compound SP318C. Same procedure as above with Trp-NHPh-OH 149c (104.4 mg, 0.353 mmol), N-Boc-Tyr(Bn)-Asn-OH 147c (171.3 mg, 0.353 mmol), EDC (74.9 mg, 0.39 mmol) and HOBt (52.6 mg, 0.39 mmol) in CH ₂ Cl ₂ ZDMF (2.2 mL, 1/1). The crude residue was triturated with CH ₂ Cl ₂ /ρentane and the resulting solid was purified by flash column chromatography on silica gel (5-8% MeOH/CH ₂ Cl ₂ ) to give an off-white solid (118.9 mg, 44 %). ¹ H NMR (300 MHz, DMSO- d ₆ ) δ 1.28 (s, 9H ₃ (CHa) ₃ ), 2.57-3.24 (m, 6H, CH ₂ Tyr, CH ₂ Trp, CH ₂ Asn), 4.11 (m, IH, CHa), 4.55 (m, 2H, 2 CHa), 5.03 (s, 2H, CH ₂ (Bn)), 6.65-7.57 (m, 21H), 8.16 (m, 2H), 9.16 (s, IH), 9.56 (s, IH), 10.76 (s, IH) . ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 27.4 (CH ₂ Trp), 28.1 ((CHj) ₃ ), 36.4, 37 (CH ₂ Tyr, CH ₂ Asn), 49.5, 54.3, 55.7 (CHa Tyr, Asn, Trp), 69.1 (CH ₂ (Bn)), 78.1 (C(CHs) ₃ ), 109.9, 111.2, 114.2, 114.8, 118.2, 120.8, 121.3, 123.5, 127.2, 127.5, 127.7, 128.3, 130.1, 130.2, 130.3, 130.4, 136, 137.2 (24 aromatic C), 153.4, 155.2, 156.8 (2 Car-O, CO carbamate), 169.1, 170.8, 171.6, 171.9 (4 CO amide). Anal. Calcd. for C ₄₂ H ₄₆ N ₆ O ₈ , 1 H ₂ O: C, 64.60; H, 6.19; N, 10.76. Found: C, 64.84; H, 6.00; N, 11.68.

N-Boc-Tyr(Bn)-Arg(Z) _r Trp-NH(4~OH)Ph . compound SP320P2. Same procedure as above with Trp-NHPhOH (112 mg, 0.357 mmol), N-Boc-Tyr(Bn)-Arg(Z) ₂ -OH (282 mg, 0.355 mmol), EDC (74.8 mg, 0.389 mmol) and HOBt (53.3 mg, 0.394 mmol) in CH ₂ Cl ₂ ZDMF

(2.2 mL, 1/1). The crude residue (235.8 mg) was purified by flash column chromatography on silica gel (1-2% MeOH/CH ₂ Cl ₂ ) to give a yellowish solid (76.2 mg, 20%). ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.25 (s, 9H, (CH ₃ ) ₃ ), 1.62 (m, 4H, 2 CH ₂ Arg), 2.6-3.2 (m, 4H, CH ₂ Tyr, CH ₂ Trp), 3.83 (m, 2H, CH ₂ Arg), 4.11 (m, IH, CHa) ₅ 4.35 (m, IH, CHa), 4.65 (m, IH, CHa), 5.01 (s, 4H, CH ₂ (Bn), CH ₂ (Z)), 5.17 (m, 2H, CH ₂ (Z)), 6.63 (d, J = 8.8 Hz ₅ 2H), 6.85 (m, 2H), 6.93 (t, J = 7.4 Hz, IH), 7.02 (t, J = 7.4 Hz ₅ IH) ₅ 7.12 (m, 2H) ₅ 7.28-7.42 (m, 20H), 7.59 (d, J = 7.7 Hz ₅ IH), 7.95 (d, J = 8 Hz ₅ IH) ₅ 8.18 (d, J = 7.8 Hz, IH), 9.15 (broad s, 3H), 9.74 (s, IH), 10.8 (s, IH) . ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 24.9, 29.7 (2 CH ₂ Arg), 27.8 (CH ₂ TrP) ₅ 28 ((CH ₃ ) ₃ ), 36.3 (CH ₂ Tyr), 44.3 (CH ₂ Arg), 52.3, 54, 55.8 (CHa Tyr, Arg, Tip), 66.1, 68.1, 69 (CH ₂ (Bn), 2 CH ₂ (Z)), 78 (C(CH ₃ ) ₃ ), 109.6, 111.2, 114.2, 114.9, 118.1, 118.4, 120.8, 121.1, 123.5, 127.3, 127.5, 127.6, 127.7, 127.8, 128.2, 128.3, 128.4, 128.5, 130.1, 130.3, 135.2, 136, 137, 137 (36 aromatic C), 153.3, 155 155.2, 156.8, 159.6, 162.9 (2 Car-O, 3 CO carbamate, imine), 169.2, 171.1, 171.6 (3 CO amide). Anal. Calcd. for C ₆₀ H ₆₄ N ₈ O ₁₁ , 4.5 H ₂ O: C, 62.43; H, 6.37; N, 9.7. Found: C, 62.38; H, 5.82; N, 9.85.

N-Boc-Tyr(Bn)-Lys(Boc)-Trp-NH(4-OH)Ph : compound SP319P. Same procedure as above with Trp-NHPhOH (75 mg, 0.239 mmol), N-Boc-Tyr(Bn)-Lys(Boc)-OH (143 mg, 0.238 mmol), EDC (50.6 mg, 0.264 mmol) and HOBt (35.3 mg, 0.261 mmol) in CH ₂ Cl ₂ ZDMF (1.6 mL, 1/1). The crude residue was triturated with MeOH/pentane to afford a white solid (142.5 mg, 68%). ¹ H NMR (300 MHz, DMSOd ₆ ) δ 1.21-1.61 (m, 6H, 3 CH ₂ Lys), 1.29 (s, 9H, (CH ₃ ) ₃ ), 1.36 (s, 9H, (CH ₃ ) ₃ ), 2.64 (m, IH, CH ₂ Tyr), 2.86 (m, 3H, CH ₂ Lys, CH ₂ Tyr), 3.03 (dd, J = 14.7 Hz, J = 7.4 Hz, IH, CH ₂ Tip), 3.17 (dd, J = 14.7 Hz, J = 6 Hz, IH, CH ₂ Tip), 4.10 (m, IH, CHa), 4.28 (m, IH, CHa), 4.64 (m, IH, CHa), 5.02 (s, 2H, CH ₂ (Bn)), 6.65 (d, J = 8.7 Hz ₅ 2H), 6.72 (m, IH), 6.88 (d, J = 8.2 Hz, 2H), 6.94 (t, J = 7.5 Hz, IH), 7.03 (t, J = 7.5 Hz, IH) ₅ 7.13-7.42 (m, 12H), 7.6 (d, J = 7.8 Hz, IH) ₅ 7.9 (d, J = 8 Hz, IH), 8.12 (d, J = 7.3 Hz ₅ IH) ₅ 9.17 (s, IH) ₅ 9.73 (s, IH), 10.8 (s, IH) . ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 22.5, 29.2, 32 (3 CH ₂ Lys), 27.8 (CH ₂ Tip), 28.1, 28.2 (2 (CH ₃ ) ₃ ), 36.3 (CH ₂ Tyr), 39.7 (CH ₂ Lys), 52.4, 54, 55.8 (CHa Tyr, Lys, Tip), 69 (CH ₂ (Bn)), 77.3, 78.1 (2 C(CH ₃ ) ₃ ), 109.6, 111.2, 114.2, 115, 118.2, 118.4, 120.8, 121.1, 123.4, 127.3, 127.5, 127.7, 128.3, 130.1, 130.2, 130.4, 136, 137.2 (24 aromatic C), 153.3, 155.2, 155.5, 156.8 (2 Car-O, 2 CO carbamate),

169.3, 171.4, 171.6 (3 CO amide). Anal. Calcd. for C ₄₉ H ₆₀ N ₆ O ₉ , 3 H ₂ O: C, 63.20; H, 7.14; N, 9.02. Found: C, 63.02; H, 6.75; N, 9.21.

N-Boc-Tyr(Bn)-Ala-Trp-NH(CH ₂ ) ₄ NHBoc : compound SP305R. Same procedure as above with Trp-NH(CH ₂ ) ₄ NHBoc (204.6 mg, 0.546 mmol), N-Boc-Tyr(Bn)-Ala-OH (242 mg, 0.546 mmol), EDC (116 mg, 0.61 mmol) and HOBt (82.2 mg, 0.61 mmol) in CH ₂ C1 ₂ /DMF (2.2 mL, 1/1). The crude tripeptide (282 mg) can be recristallized from hot THF to give a white solid as an analytical sample (89.1 mg, 31.5%). ¹ H NMR (300 MHz, DMSO- d ₆ ) δ 1.18-1.4 (m, 7H, 2 CH ₂ putrescine, CH ₃ ), 1.28 (s, 9H ₅ (CH ₃ ) ₃ ), 1.37 (s, 9H, (CH ₃ ) ₃ ), 2.61 (m, IH, CH ₂ Tyr or Tip), 2.85-3.1 (m, 7H, CH ₂ Tyr, CH ₂ Trp, 2 CH ₂ putrescine), 4.08 (m, IH, CHa), 4.29 (m, IH, CHa), 4.45 (m, IH, CHa), 5.03 (s, 2H, CH ₂ (Bn)), 6.73 (m, IH), 6.87-7.43 (m, 14 aromatic H), 7.56 (d, J = 7.7 Hz, IH), 7.83 (m, IH), 7.99 (m, 2H), 10.8 (s, INH). ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 18.2 (CH ₃ ), 26.2, 26.8 (2 CH ₂ putrescine), 28.1 (CH ₂ Trp), 28.2 ((CH ₃ ) ₃ ), 36.3 (CH ₂ Tyr), 38.2, 39.2 (2 CH ₂ putrescine), 48.2, 53.4, 55.8 (CHa Ala, Tyr, Trp), 69 (CH ₂ (Bn)), 77.3, 78 (2 C(CH ₃ ) ₃ ), 109.8, 111.1, 114.2, 118.1, 118.4, 120.7, 123.4, 127.3, 127.5, 127.7, 128.3, 130.1, 130.3, 136, 137.2 (19 aromatic C), 155.2, 155.5, 156.8 (Car-O, 2 CO carbamate), 170.7, 171.4, 171.8 (3 CO amide). Anal. Calcd. for C ₄₄ H ₅₈ N ₆ O ₈ : C, 66.14; H, 7.32; N, 10.52. Found: C, 65.89; H, 7.34; N, 10.77.

N-Boc-Tyr(Bn)-Leu-Trp-NH(CH ₂ ) ₄ NHBoc : compound SP296P. Same procedure as above with Trp-NH(CH ₂ ) ₄ NHBoc (264.7 mg, 0.706 mmol), N-Boc-Tyr(Bn)-Leu-OH (342.6 mg, 0.707 mmol), EDC (150.3 mg, 0.784 mmol), HOBt (105.3 mg, 0.78 mmol) and NEt ₃

(0.39 mL, 2.8 mmol) in CH ₂ C1 ₂ /DMF (3 mL, 1/1). The crude residue was triturated with Et ₂ O/pentane to give a white solid (196.7 mg, 33%). ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 0.82 (d, J = 6.2 Hz, 3H, CH ₃ Leu), 0.86 (d, J = 6.2 Hz, 3H, CH ₃ Leu), 1.25-1.41 (m, 4H, 2 CH ₂ putrescine), 1.29 (s, 9H, (CH ₃ ) ₃ ), 1.36 (s, 9H, (CH ₃ ) ₃ ) ₅ 2.59-3.1 (m, 8H, CH ₂ Tyr, CH ₂ Trp, 2 CH ₂ putrescine), 4.09 (m, IH, CHa), 4.32 (m, IH, CHa), 4.44 (m, IH, CHa), 5.02 (s, 2H, CH ₂ (Bn)), 6.71 (m, IH), 6.87-7.4 (m, 14 aromatic H), 7.54 (d, J = 7.7 Hz, IH), 7.79 (m, IH), 7.94 (m, 2H), 10.8 (s, INH). ¹³ C NMR (75 MHz, DMSO-(J ₆ ) δ 20.3, 21.8, 22.7 (2 CH ₃ , CH Leu), 24.9, 25.5 (2 CH ₂ . putrescine), 26.7 (CH ₂ Trp), 26.9, 27 (2 (CH ₃ ) ₃ ), 34.9 (CH ₂ Tyr), 37, 39.2, 39.7 (2 CH ₂ putrescine, CH ₂ Leu), 49.8, 52.2, 54.6 (CHa Ala, Tyr, Trp), 67.8 (CH ₂ (Bn)), 76, 76.8 (2 C(CH ₃ ) ₃ ), 108.6, 109.9, 113, 116.9, 117.1, 119.5, 122.1, 126.1, 126.3, 126.5, 127.1, 128.9, 129.1, 134.7, 136 (19 aromatic C), 154, 154.3, 155.6 (Car-O, 2 CO carbamate), 169.5, 170.3, 170.4 (3 CO amide). Anal. Calcd. for C ₄₇ H ₆₄ N ₆ O ₈ , 0.5 H ₂ O: C, 66.40; H, 7.70; N, 9.88. Found: C, 66.34; H, 7.67; N, 10.07.

,99

N-Boc-Tyr(Bn)-Asn-Trp-NH(CH ₂ ) ₄ NHBoc : compound SP323C2. Same procedure as above with Trp-NH(CH ₂ ) ₄ NHBoc (108.7 mg, 0.29 mmol), N-Boc-Tyr(Bn)-Asn-OH (140.9 mg, 0.29 mmol), EDC (61.8 mg, 0.32 mmol) and HOBt (43.6 mg, 0.32 mmol) in CH ₂ Cl ₂ ZDMF (2 mL, 1/1). The crude residue was purified by flash column chromatography on silica gel (2-6% MeOH/CH ₂ Cl ₂ ) to give an white solid (69.3 mg, 28 %). ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.18-1.37 (m, 4H, 2 CH ₂ putrescine), 1.28 (s, 9H, (CH ₃ ) ₃ ), 1.37 (s, 9H, (CH ₃ ) ₃ ), 2.42-3.2 (m, 1OH, CH ₂ Tyr, CH ₂ Trp, CH ₂ Asn, 2 CH ₂ putrescine), 4.1 (m, IH, CHa), 4.36 (m, IH, CHa), 4.53 (m, IH, CHa), 5.03 (s, 2H, CH ₂ (Bn)), 6.71 (m, IH), 6.8- 7.53 (m, 17H), 7.84 (m, IH), 8.01 (d, J = 9 Hz, IH), 8.13 (d, J = 7.6 Hz, IH), 10.73 (s, IH) . ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 26.4, 27 (2 CH ₂ putrescine), 27.4 (CH ₂ Trp), 28.4 ((CH ₃ ) ₃ ), 36.6, 37.2 (CH ₂ Tyr, CH ₂ Asn), 38.4, 39.4 (2 CH ₂ putrescine), 49.8, 54, 55.9 (CHa Tyr, Asn, Trp), 69.2 (CH ₂ (Bn)), 77.5, 78.2 (C(CH ₃ ) ₃ ), 110.2, 111.3, 114.4, 118.3, 120.9, 123.6, 127.4, 127.7, 127.9, 128.5, 130.3, 130.4, 136.1, 137.4 (19 aromatic C), 155.4, 155.7, 157 (Car-O, 2 CO carbamate), 170.7, 170.9, 171.8, 172.1 (4 CO amide). Anal. Calcd. for C ₄₅ H ₅₉ N ₇ O ₉ : C, 64.19; H, 7.06; N, 11.64. Found: C, 63.97; H, 7.05; N, 11.76.

N-Boc-Tyr(Bn)-Arg(Z) ₂ -Trp-NH(CH ₂ ) ₄ NHBoc : compound SP311C. Same procedure as above with Trp-NH(CH ₂ ) ₄ NHBoc (105.7 mg, 0.282 mmol), N-Boc-Tyr(Bn)-Arg(Z) ₂ -OH (224.5 mg, 0.282 mmol), EDC (59.5 mg, 0.31 mmol) and HOBt (42.5 mg, 0.31 mmol) in CH ₂ Cl ₂ ZDMF (1.8 mL, 1/1). The crude residue was purified by flash column chromatography on silica gel (0-2% MeOH/CH ₂ Cl ₂ ) to give a white solid (191 mg, 58.7%). ¹ H NMR (300 MHz, DMSO-dβ) δ 1.11-1.35 (m, 4H, 2 CH ₂ putrescine), 1.26 (s, 9H, (CH ₃ ) ₃ ), 1.35 (s, 9H, (CHa) ₃ ), 1.6 (m, 4H, 2 CH ₂ Arg), 2.62 (m, IH, CH ₂ Tyr), 2.82-3.06 (m, 7H, CH ₂ Tyr, CH ₂ Trp, 2 CH ₂ putrescine), 3.84 (m, 2H, CH ₂ Arg), 4.09 (m, IH, CHa), 4.31 (m, IH ₅ CHa), 4.46 (m, IH, CHa), 5.02 (broad s, 4H, CH ₂ (Bn), CH ₂ (Z)), 5.23 (s, 2H ₅ CH ₂ (Z)), 6.69 (m ₅ IH) ₅ 6.85-7.55 (m, 26H), 7.8 (m, IH) ₅ 7.97 (m, 2H) ₅ 9.16 (broad s, 2H), 10.76 (s, IH) . ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 24.8, 29.6 (2 CH ₂ Arg), 26.1, 26.7 (2 CH ₂ putrescine), 27.6 (CH ₂ Trp), 28, 28.2 ((CH ₃ ) ₃ ), 36.3 (CH ₂ Tyr), 38.2, 39.4 (2 CH ₂ putrescine), 44.3 (CH ₂ Arg), 52.3, 53.5, 55.9 (CHa Tyr, Arg, Trp), 66.1, 68.2, 69 (CH ₂ (Bn), 2 CH ₂ (Z)), 77.3, 78 (C(CH ₃ ) ₃ ), 109.7, 111.1, 114.2, 118.1, 118.4, 120.8, 123.4, 127.3, 127.4, 127.5, 127.7, 127.8, 127.9, 128.2, 128.3, 128.4, 128.5, 130.1, 130.2, 135.2, 135.9, 136, 137, 137.1 (31 aromatic C), 155, 155.1, 155.3, 159.6, 162.9 (Car-O, 3 CO carbamate, CO imine), 170.7, 171, 171.6 (3 CO amide). Anal. Calcd. for C ₆₃ H ₇₇ N ₉ O ₁₂ , 1 H ₂ O: C, 64.65; H, 6.80; N, 10.77. Found: C, 64.81; H, 6.63; N, 10.54.

N-Boc-Tyr(Bn)-Lys(Boc)-Trp-NH(CH ₂ ) ₄ NHBoc : compound SP308P. Same procedure as above with Trp-NH(CH ₂ ) ₄ NHBoc (152.4 mg, 0.407 mmol), N-Boc-Tyr(Bn)-Lys(Boc)-OH

(247.3 mg, 0.407 mmol), EDC (85.8 mg, 0.45 mmol) and HOBt (60.7 mg, 0.45 mmol) in CH ₂ C1 ₂ /DMF (1.8 mL, 1/1). The crude residue was triturated with MeOH/pentane to afford a white solid (255.9 mg, 65%). ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.29-1.57 (m, 1OH, 3 CH ₂ Lys, 2 CH ₂ putrescine), 1.29 (s, 9H, (CH ₃ ) ₃ ), 1.36 (s, 9H, (CH ₃ ) ₃ ), 2.64 (m, IH, CH ₂ Tyr), 2.86-3.11 (m, 9H, CH ₂ Lys, CH ₂ Tyr, CH ₂ Trp, 2 CH ₂ putrescine), 4.10 (m, IH, CHa), 4.26 (m, IH, CHa), 4.46 (m, IH, CHa), 5.03 (s, 2H, CH ₂ (Bn)), 6.70 (broad s, IH), 6.87-7.4 (m, 15H), 7.56 (d, J = 7.6 Hz, IH), 7.81-7.97 (m, 3H), 10.8 (s, IH) . ¹³ C NMR (75 MHz, DMSO- d ₆ ) δ 22.4, 29.2, 32 (3 CH ₂ Lys), 26.2, 26.7 (2 CH ₂ putrescine), 27.8 (CH ₂ Trp), 28, 28.1, 28.2 (3 (CH ₃ ) ₃ ), 36.3 (CH ₂ Tyr), 38.2, 39.5, 39.8 (2 CH ₂ putrescine, CH ₂ Lys), 52.4, 53.4, 55.8 (CHa Tyr, Lys, Trp), 69 (CH ₂ (Bn)), 77.2, 78 (2 C(CH ₃ ) ₃ ), 109.8, 111.1, 114.2, 118.1, 118.4, 120.7, 123.4, 127.3, 127.5, 127.6, 128.3, 130.1, 135.9, 137.1 (19 aromatic C), 154.7, 155.5, 156.7 (Car-O, 2 CO carbamate), 170.7, 171.1, 171.5 (3 CO amide). Anal. Calcd. for C ₅₂ H ₇₃ N ₇ O ₁₀ : C, 65.32; H, 7.70; N, 10.25. Found: C, 65.30; H, 7.64; N, 10.04.

N-Boc-Tyr-Arg-Trp-NHCH ₂ Ph : compound SP325. A solution of N-Boc-Tyr(Bn)- Arg(Z ₂ )-Trp-NHCH ₂ Ph (86.15 mg, 0.0804 mmol) in MeOH/DMF (1.4 mL, 1/0.4) was hydrogenated at atmospheric pressure over 10% Pd on charcoal (9.14 mg) for 18 h. The mixture was filtered over a pad of celite and concentrated. The resulting residue was triturated in Et ₂ O and filtered to afford the product as a white solid (46.55 mg, 81 %). ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.09-1.63 (m, 4H, (CH ₂ ) ₂ Arg), 1.30 (s, 9 H, (CH ₃ ) ₃ ), 2.62-3.14 (m, 6H), 4.05 (m, IH, CHa), 4.23 (m, 3H, CH ₂ Ph and CHa), 4.56 (m, IH, CHa), 6.60 (m, 2H, aromatic CH), 6.96-7.58 (m, HH, aromatic H), 7.96 (m, IH, aromatic H). ¹³ C NMR (75 MHz, CD ₃ OD) δ 25.9 (CH ₂ γ Lys), 28.7 (CH ₃ Boc), 28.9, 29.9(CH ₂ Trp or CH ₂ β Lys), 38.1 (CH ₂ Tyr), 41.9 (CH ₂ N Lys), 44.2 (CH ₂ NPh), 54.3, 55.9, 57.9 (CH α Lys or Trp or Tyr), 80.9 (C Boc), 110.7 (Car), 112.4, 118.0, 119.4, 119.9, 122.5, 124.6 (CH ar), 126.0 (Car), 128.1, 128.4 (CH ar), 128.8 (Car), 129.4, 131.3 (CH ar), 138.0, 139.4 (Car), 157.9, 158.6, 161.5 (C=N or CarO or NCOO), 173.3, 173.8, 175.0 (CONH). HRMS (ESI) calcd for C ₃₈ H ₄₉ N ₈ O ₆ [(M+H) ⁺ ] 713.3775, found 713.3778.

N-Boc-Tyr-Arg-Trp-NH(4-OH)Ph : compound SP324. Same procedure as above using N-Boc-Tyr(Bn)-Arg(Z ₂ )-Trp-NH(4-OH)Ph (60.81 mg, 0.0567 mmol), 10% Pd on charcoal (6.43 mg) in MeOH/DMF (0.8 mL, 7/1) and affording N-Boc-Tyr-Arg-Trρ-NH(4-OH)Ph as a beige solid (34.89 mg, 86 %). ¹ H NMR (300 MHz ₅ DMSO-d ₆ ) δ 1.23-1.65 (m, 4H, (CH ₂ ) ₂ Arg), 1.31 (s, 9 H, (CH ₃ ) ₃ ), 2.73-3.17 (m, 6H), 4.09 (m, IH, CHa), 4.28 (m, IH, CHa), 4.63 (m, IH, CHa), 6.59 (m, 3H, aromatic CH), 6.92-7.31 (m, 7H, aromatic H), 7.57 (d, J = 7.6 Hz, IH, aromatic H). ¹³ C NMR (75 MHz, CD ₃ OD) δ 26.1 (CH ₂ γ Lys), 28.7 (CH ₃ Boc), 26.7, 28.9(CH ₂ Trp or CH ₂ β Lys), 34.8 (CH ₂ Tyr), 41.9 (CH ₂ N Lys), 53.7, 56.4, 57.6 (CH α Lys or Trp or Tyr), 80.8 (C Boc), 110.6 (Car), 112.3, 116.9 (CH ar), 117.2 (C ar), 119.4, 119.9, 122.4, 124.1, 124.6 (CH ar), 128.7 (Car), 131.3 (CH ar), 138.0 (Car), 157.8, 158.4 (C=N or CarO or NCOO), 172.2, 173.3, 174.7 (CONH). HRMS (ESI) calcd for C ₃₇ H ₄₇ N ₈ O ₇ [(M+H) ⁺ ] 715.3568, found 715.3572.

N-Boc-Tyr(Bn)-Gly-Trp-OH : compound NR35. Saponification of N-Boc-Tyr(Bn)-Gly- Trp-OMe (242 mg, 0.385 mmol) in THF (0.5 mL) by 1 M aqueous LiOH (0.5 mL, 0.5 mmol) afforded after acidic treatment and precipitation of the residue with CH ₂ Cl ₂ /pentane N-Boc- Tyr(Bn)-Gly-Trp-OH as a white solid. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.28 (s, 9H, (CH ₃ ) ₃ ), 2.63 and 3.01 (ABX system, 2H, CH ₂ β), 3.05 and 3.17 (ABX system, 2H, CH ₂ β), 3.73 (m, 2H, CH ₂ GIy), 4.10 (m, IH, CHa), 4.49 (m, IH, CHa), 5.04 (broad s, 2H, CH ₂ O), 6.89 (m, 3H), 6.98 (t, J = 7.1 Hz, IH), 7.06 (t, J = 6.8 Hz, IH), 7.15 (m, 3 H), 7.35 (m, 6 H), 7.42 (d, J = 6.5 Hz, IH), 8.11 (m, 2H), 10.86 (s, IH). ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 27.6 (CH ₂ Trp), 28.5 (CH ₃ ) ₃ ), 37.0 (CH ₂ Tyr), 42.2 (CH ₂ αGly), 53.4, 56.3 (CHa Tyr or Tip), 69.5 (CH ₂ (OBn)), 78.4 (C Boc), 110.0 (aromatic C), 111.7, 114.7, 118.5, 118.7, 121.3, 124.1

(aromatic CH), 127.6 (aromatic C), 127.9, 128.1, 128.8, 130.6 (aromatic CH) 130.7, 136.4, 137.6 (aromatic C), 155.7, 157.2 (C _ar -O, CO carbamate), 168.9, 172.4, 172.5 (2 CO amide and COOH). Anal. Calcd. for C ₃₄ H ₃₈ N ₄ O ₇ , 1 H ₂ O: C, 64.54; H, 6.37; N, 8.85. Found: C ₅ 64.48; H, 6.35; N, 8.70.

N-Boc-Tyr(Bn)-Ala-Trp-OCH ₃ : compound NR36. Same procedure as above with HCl, Trp-OCH ₃ (255 mg, 1 mmol), N-Boc-Tyr(Bn)-Ala-OH (444 mg, 1 mmol), DCC (259 mg, 1.26 mmol) and HOBt (172 mg, 1.27 mmol) in THF (15 mL). The crude residue was chromatographed over silica gel to afford a white solid (361 mg, 56%). ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.22 (d, J = 7 Hz, 3H, CH ₃ ), 1.29 (s, 9H, (CHa) ₃ ), 2.62 and 2.92 (ABX system, 2H, CH ₂ β), 3.08 and 3.13 (ABX system, 2H, CH ₂ β) 3.55 (s, 3H, OCH ₃ ), 4.10 (m, IH, CHa), 4.36 (m, IH, CHa), 4.49 (m, IH ₅ CHa), 5.03 (s, 2H, CH ₂ (OBn)), 6.85-7.50 (m, 15 H, 14 aromatic H + 1 NH), 7.97 (d, J = 7.3 Hz, IH), 8.34 (d, J = 7.0 Hz, IH), 10.88 (s, INH). ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 18.2 (CH ₃ ), 26.8 (CH ₂ Tip), 28.0 ((CH ₃ ) ₃ ), 36.3 (CH ₂ Tyr), 47.6, 51.6, 53.0, 55.6 (CHa Ala, Tyr, Trp or OCH ₃ ), 69 (CH ₂ (OBn)), 77.9 (C(CH ₃ ) ₃ ), 109.0 (aromatic C), 111.2, 114.2, 117.8, 118.3, 120.8, 123.6 (aromatic CH), 126.9(aromatic C), 127.4, 127.6, 128.2, 130.0 (aromatic CH), 130.2, 135.9, 137.1 (aromatic C), 155.1, 156.7 (Car-0 or CO carbamate), 171.2, 171.9, 172.1 (2 CO amide or CO ester). Anal. Calcd. for C ₃₆ H ₄₂ N ₄ O ₇ , 0.5 H ₂ O: C, 66.34; H, 6.65; N, 8.60. Found: C, 66.29; H, 6.64; N, 8.48.

N-Boc-Tyr-Ala-Trp-OCH ₃ : compound NR40. N-Boc-Tyr(Bn)-Ala-Trp-OMe (119 mg, 0.185 mmol) in solution in MeOH (4 mL) was hydrogenated overnight at atmospheric

pressure in the presence of 10% Pd on charcoal (30 mg). After fϊltation, evaporation of the solvant and trituration with ether, N-Boc-Tyr-Ala-Trp-OMe (77 mg, 76 %) was obtained as a solid. ¹ H NMR (300 MHz ₅ DMSO-d ₆ ) δ 1.19 (d, J = 7 Hz, 3H ₅ CH ₃ ), 1.29 (s, 9H, (CHs) ₃ ), 2.55 and 2.80 (ABX system, 2H, CH ₂ β), 3.13 and 3.16 (ABX system, 2H, CH ₂ β), 3.55 (s, 3H, OCH ₃ ), 4.05 (m, IH, CHa), 4.35 (m, IH, CHa), 4.50 (m, IH, CHa), 6.63 (d, J = 8.3 Hz, 2H), 6.83 (d, J = 8.6 Hz, IH), 7.03 (m, 4H), 7.16 (d, J = 2.1 Hz, IH), 7.34 (d, J - 7.9 Hz, IH), 7.47 (d, J = 7.8 Hz, IH), 7.92 (d, J = 7.4 Hz, IH, NH), 8.32 (d, J = 7.3 Hz, IH, NH), 9.14 (s, IH ₅ OH) ₅ 10.86 (s, INH). ¹³ C NMR (75 MHz, DMSOd ₆ ) δ 18.4 (CH ₃ ), 26.9 (CH ₂ Trp) ₅ 28.1 ((CH ₃ ) ₃ ), 36.4 (CH ₂ Tyr), 47.8, 51.7, 53.0, 55.9 (CHa Ala, Tyr, Trp or OCH ₃ ), 77.9 (C(CH ₃ ) ₃ ), 109.1 (aromatic C), 111.4, 114.8, 117.9, 118.4, 120.9, 123.7 (aromatic CH), 127.0, 128.2 (aromatic C), 130.0 (aromatic CH), 136.0 (aromatic C), 155.2, 156.6 (Car-0 or CO carbamate), 171.4, 172.O ₅ 172.2 (2 CO amide or CO ester). Anal. Calcd. for C ₂₉ H ₃₆ N ₄ O ₇ , 1 H ₂ O: C, 61.04; H ₅ 6.71; N ₅ 9.82. Found: C, 61.02; H, 6.62; N ₅ 9.66. HRMS (ESI) calcd for C ₂₉ H ₃₆ N ₄ O ₇ Na [(M+Na) ⁺ ] 575.2482, found 575.2480.

N-Boc-Tyr(Bn)-Ala-Trp-OH : compound NR66. Saponification of N-Boc-Tyr(Bn)-Ala- Trp-OMe (580 mg, 0.902 mmol) in THF (5 mL) by 1 M aqueous LiOH (2 mL, 2 mmol) at 4 ⁰ C for 1 hour, afforded after acidic treatment and precipitation of the residue with water crude N-Boc-Tyr(Bn)-Gly-Trp-OH. The crude product in MeOH (0.5 mL) was treated by dicyclohexylamine (0.18 mL, 0.91 mmol) and then ether (10 mL). After filtration, the resulting solid was dissolved in CH ₂ Cl ₂ (10 mL) and washed by 10% aqueous citic acid. The organic phase was dried over MgSO4 and concentrated to afford N-Boc-Tyr(Bn)-Ala-Trp-OH (361 mg, 64 %) as a white solid. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.20 (d, J = 7 Hz, 3H, CH ₃ ), 1.28 (s, 9H, (CH ₃ ) ₃ ), 2.62 and 2.86 (ABX system, 2H, CH ₂ β), 3.09 and 3.18 (ABX system, 2H ₅ CH ₂ β), 4.10 (m, IH, CHa), 4.35 (m, IH, CHa), 4.46 (m, IH, CHa) ₅ 5.03 (s, 2H, CH ₂ (OBn)), 6.87-7.54 (m, 15 H, 14 aromatic H + 1 NH), 7.96 (d, J = 7.5 Hz, IH) ₅ 8.13 (d, J = 7.5 Hz, IH), 10.84 (s, INH). ¹³ C NMR (75 MHz, CDCl ₃ ) δ 18.2 (CH ₃ ), 27.2 (CH ₂ Trp), 28.3 ((CHs) ₃ ), 37.3 (CH ₂ Tyr), 48.9, 53.3, 55.7 (CHa Ala, Tyr or Trp), 70.0 (CH ₂ (OBn)), 80.7 (C(CHa) ₃ ), 109.5 (aromatic C), 111.5, 115.0, 118.6, 119.5, 121.9, 123.8, 127.5, 127.6,

128.1, 128.6, 130.4 (aromatic CH), 136.1, 137.0, 137.1 (aromatic C), 155.8, 157.8 (Car-0 or CO carbamate), 171.9, 172.3, 174.2 (2 CO amide or CO acid). Anal. Calcd. for C ₃₅ H ₄₀ N ₄ O ₇ , 1.5 H ₂ O: C, 64.11% ; H, 6.61% ; N,8.54%. Found: C, 64.19%; H, 6.36%; N,8.79%.

N-Boc-Tyr-Ala-Trp-OH : compound NR68. N-Boc-Tyr(Bn)-Ala-Trp-OH (233 mg, 0.37 mmol) in solution in MeOH (4 mL) was hydrogenated overnight at atmospheric pressure in the presence of 10% Pd on charcoal (40 mg). After filtration, evaporation of the solvent and trituration with ether, N-Boc-Tyr-Ala-Trp-OMe was obtained as a solid (152 mg, 76 %). H NMR (300 MHz, DMSO-d ₆ ) δ 1.21 (d, J = 7 Hz, 3H, CH ₃ ), 1.29 (s, 9H ₅ (CH ₃ ) ₃ ), 2.51 and 2.83 (ABX system, 2H, CH ₂ β), 3.05 and 3.20 (ABX system, 2H, CH ₂ β), 4.05 (m, IH, CHa), 4.33 (m, IH, CHa), 4.42 (m, IH, CHa), 6.62 (d, J = 8.3 Hz, 2H), 6.83 (d, J = 8.6 Hz, IH), 7.0 (m, 4H), 7.14 (d, J = 2.1 Hz, IH), 7.31 (d, J = 7.9 Hz, IH), 7.52 (d, J = 7.7 Hz, IH), 7.93 (d, J = 7.5 Hz, IH, NH), 8.09 (d, J = 7.4 Hz, IH, NH), 9.13 (s, IH, OH), 10.82 (s, INH). ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 18.4 (CH ₃ ), 27.0 (CH ₂ Tip), 28.1 ((CHs) ₃ ), 36.4 (CH ₂ Tyr), 47.9, 53.0, 55.9 (CHa Ala, Tyr or Tip), 78.0 (C(CHa) ₃ ), 109.6 (aromatic C), 111.3, 114.8, 118.1, 118.3, 120.8, 123.6 (aromatic CH), 127.1, 127.2 (aromatic C), 130.0 (aromatic CH), 136.0 (aromatic C), 155.2, 157.6 (Car-0 or CO carbamate), 171.4, 172.0, 173.2 (2 CO amide or CO acid). Anal. Calcd. for C ₂₈ H ₃₄ N ₄ O ₇ , 2.5 H ₂ O: C, 57.62%; H, 6.74%; N, 9.60%. Found: C, 57.66%; H, 6.57%; N, 9.45%.

VI) Preparation of tripeptides containing oxotryptophane: compounds IV-3

Hydrogenolysis of L-Z-Trp [O]-NHPh and coupling to dipeptides. General procedure applied to

Boc-Tyr(Bn) -Arg(Z) ₂ -Trp[O]-NHPh : compound CVIl. A mixture of L Z-Trp[O]

-NHPh (1.29 g, 3 mmol) and 10% palladium on charcoal (259 mg) in DMF (20 mL) and

MeOH (20 mL) was hydrogenated at atmospheric pressure for 5 h. After filtration over a pad of celite, the filtrate was concentrated and the resulting residue was washed with ether to afford L-Trp[O]-NHPh as a white solid (0.76 g, 85%).

To a solution of dipeptide Boc-Tyr(Bn)-Arg(Z) ₂ -OH (517.2 mg, 0.65 mmol) in CH ₂ Cl ₂ at 0 °C was added HOBt (97.5 mg, 0.72 mmol) and EDC, HCl (137.5 mg, 0.72 mmol). The mixture was stirred for 15 min at 0 °C before adding a solution of L-Trp[O]-NHPh (202.2 mg, 0.68 mmol) in DMF (1.5 mL). After stirring overnight at room temperature, the solvent was evaporated in vacuo. The residue was dispersed in water (3 mL), filtered and successively washed with water and Et ₂ O to afford the crude tripeptide (497 mg). After chromatography on silica gel (30 g, 4% MeOH/ CH ₂ Cl ₂ ), Boc-Tyr(Bn) -Arg(Z) ₂ -Trp[O]-NHPh was obtained as a white solid (262 mg, 37 %). ¹ H NMR (300 MHz, DMSO-d ₆ ), (64/36 mixture of diastereomers) δ 1.25 (s, 9H, (CH ₃ ) ₃ ), 1.30-1.69 (m, 4H, 2 CH ₂ (Arg)), 1.89-2.88 (m, 4H, CH ₂ Trp and CH ₂ Tyr), 3.44 (m, IH, CH oxindole), 3.87 (m, 2H, CH ₂ -N (Arg)), 4.11 (m, IH, CHo), 4.39 (m, IH, CH _α ), 4.87 (m, IH, CH ₀ O, 5.01-5.05 (m, 4H, 2 CH ₂ (Z)), 5.17 (m, 2H, CH ₂ (Bn)), 6.83-7.56 (m, 29H, 28 aromatic H and NH), 8.06-8.62 (m, 2H, 2 NH), 9.15 (broad s, 2H, NH), 9.97 (s, IH, NH), 10.39 and 10.45 (two s, IH, oxindolic NH of dia 1 or dia 2 ). ¹³ C NMR (75 MHz, DMSOd ₆ ) (mixture of diastereomers) δ 25 (CH ₂ Arg), 28 (CH ₃ Boc), 29.4 (CH ₂ Arg), 33.3 (CH ₂ Tip), 36.3 (CH ₂ Tyr), 41.9 (CHγ Trp[O]), 44.4 (CH ₂ -N Arg), 50.9, 52.5, 55.8 (3 CHa), 66.1, 68.1, 69.1 (2 CH ₂ (Z) and CH ₂ (Bn)), 77.9 (C Boc) , 109.3-142.5 (35 aromatic CH or C), 154.9 , 155.1 , 156.8 , 159.6 , 162.8 (3 CO carbamate, 1 Car-O and 1 C=NH) , 169.8 , 171.3 , 171.8 (3 CO amide), 178.4 et 178.6 (oxindolic CO of dia 1 or dia2). Anal. Calcd. for C ₆₀ H ₆₄ N ₈ O ₁₁ : C, 67.15; H, 6.01; N, 10.44. Found: C, 67.04; H, 6.00; N, 10.15.

Boc-Tyr(Bn)-Ala-Trp [O]-NHPh : compound CV13. The general procedure starting from dipeptide Boc-Tyr(Bn)- AIa-OH (144.2 mg, 0.32 mmol), HOBt (51.6 mg, 0.37 mmol), EDC, HCl (68.3 mg, 0.35 mmol) and Trp[O]-NHPh (98.7 mg, 0.35 mmol) afforded, after washing with ether (3 X 5 mL) the tripeptide Boc-Tyr(Bn)-Ala-Trp[O]-NHPh as a white solid (101.2 mg, 44%). ¹ H NMR(300 MHz, DMSO-d ₆ )(50/50 mixture of diastereomers) δ 1.28 (broad s, 12H, CH ₃ Boc and CH ₃ Ala), 2.07-2.93 (m, 4H, CH ₂ Trp[O] and CH ₂ Tyr), 3.43 (s, IH, CHγ Trp[O]), 4.13 (m, IH, CHa), 4.35 (m, IH, CHa), 4.8 (m, IH, CHa), 5.03 (s, 2H, CH ₂ (Bn)), 6.87-7.61 (m, 19H, 18 aromatic H and NHBoc), 8.10 (m, IH, NH), 8.28 and 8.53 (two d, IH, J = 7.6 Hz and J = 9.0 Hz, dia 1 or dia 2 NH Trp[O]), 9.93 and 9.97 (two s, IH, NHPh dia 1 or dia 2), 10.42 and 10.45 (two s, IH ₅ dia 1 or dia 2 NH oxindole). ¹³ C NMR (75 MHz, DMSO- d ₆ ) (mixture of two diastereomers) δ 18 (CH ₃ Ala), 28.1 (CH ₃ Boc), 33.3 (CH ₂ β Trp[O]), 36.3 (CH ₂ β Tyr), 41.9 (CHγ Trp[O]), 48.4, 51, 55.7 (3 CHa), 69.1 (CH ₂ (Bn)), 78 (C Boc), 109.3- 142.5 (23 aromatic C), 155.2 and 156.8 (1 CO carbamate et 1 Car-0), 169.9, 171.1, 172.6 (3 CO amide), 178.5 and 178.6 (CO oxindole dia 1 or dia 2). Anal. Calcd. for C ₄₁ H ₄₅ N ₅ O ₇ , 1 H ₂ O : C, 66.73; H, 6.42; N, 9.49. Found: C, 67.06; H, 6.22; N, 9.57. HRMS (ESI) calcd for C ₄₁ H ₄₅ N ₅ O ₇ Na [(M+Na) ⁺ ] 742.3217, found 742.3234.

Boc-Tyr(Bn)-Leu-Trp [O]-NHPh : compound JV602. The general procedure starting from Boc-Tyr(Bzl)-Leu-OH (109.5 mg, 0.226 mmol), HOBt ( 36.38 mg, 0.269 mmol), EDC, HCl (50.29, 0.262 mmol) and Trp[O]-NHPh (66.32 mg, 0.224 mmol) afforded the crude tripeptide as a solid which was suspended in boiling water (4 mL). After cooling to room temperature, filtration and washing with ether (3 X 5 mL), the tripeptide Boc-Tyr(Bn)-Leu- Trp[O]-NHPh was obtained as a white solid (117.9 mg, 69 %). ¹ H NMR(SOO MHz, DMSO-

d ₆ )(40/60 mixture of diastereomers dial/dia2 which equilibrates to a 70/30 mixture within a few days at room temperature) δ 0.88-1.06 (m, 6H, CH ₃ Leu), 1.23 and 1.29 (9 H, dial and dia2, CH ₃ Boc), 1.50-1.70 (m, 3H, CHγ and CH ₂ β Leu), 1.87-2.26 (m, 2H, CH ₂ β Tip[O]), 2.66-2.93 (m, 2H, CH ₂ β Tyr), 3.39-3.48 (m, IH, CH ₂ γ ¹ T-P[O]), 4.11-4.15 (m, IH, CHa Tyr), 4.33-4.45 (m, IH, CHa Leu), 4.78-4.90 (m, IH, CHa Trp[O]), 5.02 (broad s, 2H, CH ₂ O), 6.80-7.62 (m, 19 H, 18 aromatic H and NH Boc), 7.62 and 7.97 (two d, IH, J = 6.8 Hz and J = 6.8 Hz, dia 2 and dia 1 NH Leu), 8.31 and 8.58 (two d, IH, J = 6.3 Hz and J = 7.7 Hz, dia 2 and dia 1 NH Tip[O]), 9.96 (s, IH, NHPh), 10.42 and 10.46 (two s, IH, dia2 and dia 1 indolic NH). ¹³ C NMR (75 MHz, DMSO-d ₆ ) (mixture of two diastereomers) δ 21.6, 21.7, 23.0, 23.1, 24.0, 27.8, 28.0 (CH and CH ₃ ), 32.8, 33.2, 36.1, 40.8, 41.9 (CH ₂ ), 50.9, 51.1, 51.7, 55.7 (CH), 69.0 (CH ₂ ), 77.9, 78 (C), 109.3, 114.2, 119.3, 119.4, 121.2, 123.4, 124.2, 125.2, 127.6, 127.7, 128.3, 128.6, 128.9, 129.3, 130.1, 130.2, 130.3 (CH), 137.2, 138.7, 142.4, 142.5, 155.2, 156.8, 169.8, 171.8, 171.9, 172.4, 178.5, 178.6 (C). Anal. Calcd. for C ₄₄ H _5I N ₅ O ₇ , 0.5 H ₂ O : C, 68.55; H, 6.80; N, 9.07. Found: C, 68.56; H, 6.50; N, 9.25.

Boc-Tyr(Bn)-Lys(Boc)-Trp [O]-NHPh : compound CV12. The general procedure starting from dipeptide Boc-Tyr(Bn)-Lys(Boc)-OH (392.3 mg, 0.65 mmol), HOBt (98.6 mg, 0.71 mmol), EDC, HCl (138.2 mg, 0.71 mmol) and Trp[O]-NHPh (201.3 mg, 0.65 mmol) afforded, after washing with ether (3 X 5 mL) the tripeptide Boc-Tyr(Bn)-Lys(Boc)-Trp[O]- NHPh as a white solid (273.3 mg, 48%). ¹ H NMR(300 MHz, DMSO-d ₆ )(50/50 mixture of diastereomers) δ 1.29 and 1.36 (two s, 9H ₅ CH ₃ Boc), 1.51-1.56 (m, 6H, 3CH ₂ Lys), 1.70-3.03 (m, 6H, CH ₂ β Trp[O] ,CH ₂ β Tyr and CH ₂ -N (Lys)), 3.43 (m, IH, CHγ Trp[O]), 4.14 (m, IH, CHa), 4.32 (m, IH, CHa), 4.80 (m, IH, CHa), 5.02 (s, 2H, CH ₂ (Bn)), 6.63-7.63 (m, 2OH, 18 aromatic H and 2 NH Boc), 7.97 (m, IH, NH), 8.32 and 8.60 (two d, IH, J = 6.9 Hz and J = 9.2 Hz, NH Trp[O] of dia 1 or dia 2), 9.96 (s, IH, NHPh), 10.41 and 10.46 (two s, IH, indolic NH of dia 1 or dia 2). ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 24.5 (CH ₂ Lys), 30.1 and 30.2 (2 CH ₃ Boc), 31.2 and 33.7 (2 CH ₂ Lys), 35.2 (CH ₂ ¹ TIp[O]), 38.3 (CH ₂ Tyr), 41.4 (CH ₂ - NHBoc), 43.9 (CHγ Trp[O]), 53, 54.6, 57.7 (3 CHa), 71.1 (CH ₂ (Bn)), 79.3 and 80.1 (2 C Boc), 111.3-144.5 (23 aromatic C), 157.2, 157.5, 158.8 (2 CO carbamate and 1 Car-O), 171.9, 173.8, 174 (3 CO amide), 180.5 and 180.6 (CO oxindole dia 1 or dia 2). Anal. Calcd. for

C ₄₉ H ₆₀ N ₆ O ₉ , 1.5 H ₂ O : C, 65.09; H, 7.02; N, 9.29. Found: C, 65.20; H ₅ 6.71; N, 9.66. HRMS (ESI) calcd for C ₄₉ H ₆₀ N ₆ O ₉ Na [(M+Na) ⁺ ] 899.4319, found 899.4322.

WWtt..:: 719,83

Boc-Tyr(Bn)-Gly-Trp[0]-NHCH ₂ Ph : compound NR15. The general procedure starting from dipeptide Boc-Tyr(Bn)-Gly-OH (210 mg, 0.488 minol), HOBt (67.6 mg, 0.5 mmol), EDC, HCl (94.1 mg, 0.49 mmol) and Trp[O]-NHCH ₂ Ph (139 mg, 0.449 mmol) afforded, after washing with ether (3 X 5 mL) the tripeptide Boc-Tyr(Bn)-Gly-Trp[O]-NHCH ₂ Ph as a white solid. ¹ H NMR(300 MHz, DMSO-d ₆ )(70/30 mixture of diastereomers) δ 1.19 and 1.27 (broad s, 9H, CH ₃ Boc), 1.87-2.24 (m, 2H, CH ₂ Trp[O]), 2.60-2.96 (m, 2H, CH ₂ Tyr), 3.43 (m, IH, CHγ Trp[O]), 3.82 (m, 2H, CH ₂ α GIy), 4.12 (m, IH, CHa Tyr), 4.28 (broad s, 2H, CH ₂ NBn), 4.75 (m, IH, CHa Trp[O]), 5.04 (broad s, 2H, CH ₂ (Bn)), 6.83-7.44 (m, 19H, 18 aromatic H and NHBoc), 8.20 (m, IH, NH), 8.43 and 8.52 (two d, IH, J = 8.6 Hz and J = 6.0 Hz, dia 1 or dia 2 NH ¹ TTp[O]), 10.41 and 10.45 (two s, IH, dia 1 or dia 2 NH oxindole). ¹³ C NMR (75 MHz, DMSO-d ₆ ) (mixture of two diastereomers) δ 28.1 (CH ₃ Boc), 33.1 and 33.5 (CH ₂ β Trp[O]), 36.4 (CH ₂ β Tyr), 41.8 and 41.9 (CHγ Trp[O]), 42.1 and 42.2 (CH ₂ α GIy and CH ₂ NBn) _, 50.1 and 50.8 (CHa Trp[O]), 55.8 ((CHa Tyr), 69.1 (CH ₂ (OBn)), 78,1 (C Boc), 109.2 and 109.3, 114.3, 121.2 and 121.3, 124.1 and 125, 126.1-130.3 (12 aromatic CH and 1 aromatic C), 137.2, 139.2 and 139.3, 142.4 and 142.5, 155.2 and 155.3 (4 aromatic C), 156.8 (1 CO Boc), 168, 8 and 169.2, 170.9 and 171.0, 172.1 (3 CO amide dia 1 or dia 2), 178.6 and 178.7 (CO oxindole dia 1 or dia 2). Anal. Calcd. for C ₄₁ H ₄₅ N ₅ O ₇ , 1 H ₂ O : C, 66.73; H, 6.42; N, 9.49. Found: C, 66.30; H, 6.45; N, 9.97. HRMS (ESI) calcd for C ₄₁ H ₄₅ N ₅ O ₇ Na [(M+Na) ⁺ ] 742.3217, found 742.3226.

Boc-Tyr(Bn) -Arg(Z) ₂ -Trp [O] -NHCH ₂ Ph : compound NR16. The general procedure starting from dipeptide Boc-Tyr(Bn)-Arg(Z) ₂ -OH (389 mg, 0.49 mmol), HOBt (67.5 mg, 0.5 mmol), EDC, HCl (100.5 mg, 0.524 mmol) and Trp[O] -NHCH ₂ Ph (151 mg, 0.49 mmol) afforded, after chromatography over silica gel (MeOH/CH ₂ Cl ₂ 1/20) the tripeptide Boc- Tyr(Bn)- Arg(Z) ₂ -Trp[O]-NHCH ₂ Ph as a white solid (151 mg, 28 %). ¹ H NMR (500 MHz ₅ DMSOd ₆ ), (64/36 mixture of diastereomers, COSY) δ 1.24 (broad s, 9H, (CH ₃ ) ₃ ), 1.62 (m, 4H, 2 CH ₂ (Arg)), 1.89-2.30 (m, 2H, CH ₂ βTrp[O]), 2.52-2.84 (m, 2H, CH ₂ Tyr) 3.38 (m, IH, CHγ Trp[O]), 3.87 (m, 2H, CH ₂ -N (Arg)), 4.09 (m, IH, CH _α Tyr), 4.24 (m, 2H, CH ₂ NBn), 4.37 (m, IH, CH _α Arg), 4.78 (m, IH, CH _α Trp[O]), 5.03 (broad s, 4H, 2 CH ₂ (Z)), 5.22 (broad s, 2H, CH ₂ (OBn)), 6.83-7.40 (m, 29H, 28 aromatic H and NH), 8.05 and 8.12 (two d, IH, J = 8 Hz and J = 7 Hz ₅ NH Arg), 8.33 and 8.53 (two m, IH, NH Trp[O]), 8.44 and 8.48 (two m, IH, NHBn), 9.17 (brod s, 2H, NH),, 10.40 and 10.46 (two s, IH, oxindolic NH). ¹³ C NMR (75 MHz, DMSO-d ₆ ) (mixture of diastereomers) δ 24.9 (CH ₂ Arg), 28.1 (CH ₃ Boc), 29.3 (CH ₂ Arg), 33.2 and 33.7 (CH ₂ TTp[O]), 36.3 (CH ₂ Tyr), 41.9 (CHγ Trp[O]), 42.1 (CH ₂ NBn), 44.4 (CH ₂ -N Arg), 50.2, 52.6 (3 CHa), 66.1, 68.2, 69.1 (2 CH ₂ (Z) and CH ₂ (Bn)), 78.0 (C Boc), 109.3, 114.24, 121.3, 125.1-130.3 (17 signals for aromatic CH or C), 135.3, 137.1 and 137.2, 139.1 and 139.3, 142.5 and 142.6 (aromatic C), 155.0 , 155.2 , 156.8 , 159.7 , 163.0 (3 CO carbamate, 1 Car-0 or 1 C=NH), 170.9 , 171.3 , 171.8 (3 CO amide), 178.6 and 178.7 (indolic CO of dia 1 and dia2). Anal. Calcd. for C ₆₁ H ₆₆ N ₈ O ₁₁ : C, 67.39; H, 6.12; N, 10.31. Found: C, 67.15; H, 6.23; N, 10.32.

Boc-Tyr(Bn)-GIy-Trp[0]-OCH ₃ : compound NR38. The general procedure starting from dipeptide Boc-Tyr(Bn)-Gly-OH (333 mg, 0.777 mmol), HOBt (115.7 mg, 0.856 mmol), EDC,

HCl (164.4 mg, 0.857 mmol) and Trp[O]-OMe (0.778 mmol) afforded, after chromatography over silica gel (eluant AcOEt) the tripeptide Boc-Tyr(Bn)-Gly-Trp[0]-OMe as a white solid (119 mg, 24%). ¹ H NMR(300 MHz, DMSO-d ₆ )(55/45 mixture of diastereomers) δ 1.28 (broad s, 9H, CH ₃ Boc), 2.08-2.30 (m, 2H, CH ₂ Trp[O]), 2.60-2.96 (m, 2H, CH ₂ Tyr), 3.41 (m, IH, CHγ Trp[O]), 3.53 and 3.60 (two s, 3H, OCH ₃ ), 3.77 (m, 2H, CH ₂ α GIy), 4.10 (m, IH, CHa Tyr), 4.73 (m, IH, CHa Trp[O]), 5.05 (broad s, 2H, OCH ₂ (Bn)), 6.81-7.44 (m, 15H, 13 aromatic H and NHBoc), 8.18 (m, IH, NH GIy), 8.18 and 8.51 (two d, IH, J = 7 Hz and J = 8 Hz, NH Trp[O]), 10.44 and 10.46 (two s, IH, NH oxindole). ¹³ C NMR (75 MHz, CDCl ₃ ) (mixture of two diastereomers) δ 28.3 (CH ₃ Boc), 31.1 and 31.8 (CH ₂ β Trp[O]), 37.6 (CH ₂ β Tyr), 43.0 (CH ₂ α GIy), 43.4 (CHγ Trp[O]), 50.2 and 50.7 (CHa Trp[O]), 52.6 and 52.7 (OCH ₃ ), 55.9 (CHa Tyr), 70.0 (CH ₂ (OBn)), 80.3 (C Boc), 110.3 and 110.4, 115.0, 122.7 and 122.8, 123.9 and 124.5 (aromatic CH), 127.5-130.5 (7 signals for aromatic CH and 2 aromatic C), 137.1, 141.6 and 141.8 (2 aromatic C), 155.9 and 156.0, 157.8 (1 CO Boc and 1 aromatic C), 169.1 and 169.2, 171.9 and 172.0, 172.4 and 172.5 (2 CO amide and 1 CO ester), 180.1 and 180.4 (CO oxindole). Anal. Calcd. for C ₃₅ H ₄₀ N ₄ O ₈ , 1 H ₂ O : C, 63.43; H, 6.39; N, 8.45. Found: C, 63.02; H, 6.25; N, 8.87. HRMS (ESI) calcd for C ₃₅ H ₄₀ N ₄ O ₈ Na [(M+Na) ⁺ ] 667.2744, found 667.2740.

Boc-Tyr(Bn)-Ala-Trp[0]-OCH ₃ : The general procedure starting from dipeptide Boc- Tyr(Bn)-Ala-OH, HOBt, EDC, HCl and Trp[O]-OMe afforded, after chromatography over silica gel the tripeptide Boc-Tyr(Bn)-Ala-Trp [O]-OMe.

Boc-Tyr(Bn)-Ala-Trp [O]-OH: Saponification of Boc-Tyr(Bn)-Ala-Trp[O]-OMe in THF with aqueous LiOH afforded after acidification with aqueous HCl, Boc-Tyr(Bn)-Ala-Trp[O]- OH.

Boc-Tyr-AIa-Trp[0]-OCH ₃ : Stirring of a mixture of Boc-Tyr(Bn)-Ala-Trp[O]-OMe and 10% Pd / C under atmosphere of hydrogene afforded Boc-Tyr-Ala-Trp[O]-OMe

Boc-Tyr-Ala-Trp [O]-OH: Stirring of a mixture of Boc-Tyr(Bn)-Ala-Trp [O]-OH and 10% Pd / C under atmosphere of hydrogene afforded Boc-Tyr-Ala-Trp[O]-OH

VII) Preparation of homologues of halogen ated tripeptides: compounds IV-Ib

N-Boc-3-iodo-Tyr(Me)-OMe: To a stirred suspension of I ₂ (370 mg, 1.46 mmol) and Ag ₂ SO ₄ (455 mg, 1.46 mmol) in MeOH (24 mL) was added Boc-Tyr(Me)-OMe (376 mg, 1.22 mmol) at room temperature. The mixture was stirred for 1 h. The yellow solid was removed by filtration over celite and the filtrate was concentrated off. The residue was

dissolved in CHCl ₃ and washed successively with aqueous 0.1 M Na ₂ S ₂ O ₃ , water and brine. The organic layer was dried over Na ₂ SO ₄ and the solvent was evaporated under vacuum. Purification by flash chromatography on silica gel (1-5% MeOH/CH ₂ Cl ₂ ) yielded N-Boc-3- iodo-Tyr(Me)-OMe as a yellow foam (370 mg, 69%). ¹ H RMN (300 MHz, CDCl ₃ ) δ 1.45 (s, 9H, (CH ₃ ) ₃ ) ₃ 3 (m, 2H, CH ₂ ), 3.75 (s, 3H, OCH ₃ ), 3.88 (s, 3H, OCH ₃ ), 4.51 (m, IH, CH), 5 (m, IN, NHBoc), 6.76 (d, J = 8.4 Hz, IH, H5), 7.09 (dd, J = 8.4 Hz, J = 2.1 Hz, IH, H6), 7.55 (d, J = 2.1, IH, H2). ¹³ C NMR (75 MHz, CDCl ₃ ) δ 28.3 ((CH ₃ ) ₃ ), 36.2 (CH ₂ ), 52.2 (OCH ₃ ), 54.5 (CH), 56.3 (OCH ₃ ), 79.8 (C(CH ₃ ) ₃ ), 85.5 (C3), 110.8 (CH), 130.2 (C, CH), 140.1 (CH), 155 (C4), 157.1 (CO Boc), 172.1 (CO ester).

N-Boc-3-iodo-Tyr(Me)-OH: To a solution of N-Boc-3-iodo-Tyr(Me)-OMe (2.33g, 5.35 mmol) in THF (3OmL) cooled at 0°C was added a 1 M aqueous LiOH solution (5.9 mL). The mixture was stirred for 1 h 30 at 0 ⁰ C before it was quenched by 2 N aqueous HCl solution (pH = 1-2). The aqueous phase was extracted twice by CH ₂ Cl ₂ , the combined organic layers were dried over Na ₂ SO ₄ . Removing of the solvents in vacuo afforded N-Boc-3-iodo-Tyr(Me)- OH (1.96 g, 100%) as a white foam which was used in the next step without further purification. ¹ H NMR (300 MHz, CDCl ₃ ) (80 :20 mixture of rotamers) δ 1.36 and 1.41 (two s, 9H, (CHa) ₃ ), 2.83 and 3.07 (two m, 2H, CH ₂ ), 4.88 (s, 3H, OCH ₃ ), 4.33 and 4.54 (two m, IH, CH), 4.96 and 6.13 (two m, IH, NHBoc), 6.78 (d, J = 8.4 Hz, H5), 7.15 (dd, J = 8.4 Hz, J = 2.1 Hz, H6), 7.61 (d, J = 2.1 Hz, IH, H2). ¹³ C NMR (75 MHz, CDCl ₃ ) δ 28.1 and 28.1 ((CH ₃ ) ₃ ), 36.5 and 37.6 (CH ₂ ), 53.6 and 54.5 (CH), 56.4 (OCH ₃ ), 80.3 and 81.8 (C Boc), 85.9 (C3), 130.4 (CH), 130.7 (C), 140.2 (CH), 155.5 (C4), 157.1 and 156.7 (CO Boc), 175.6 (CO acid). Treatment of the crude acid with dicyclohexylamine (1.1 eq.) in Et ₂ O gave an analytical sample of the dicyclohexylamine salt. Anal. Calcd. for C ₂₇ H ₄₃ N ₂ O ₅ I, 0.5 H ₂ O: C, 53.03; H, 7.25; N, 4.58. Found: C, 53.30; H, 7.10; N, 4.51.

N-Boc-3-iodo-Tyr(Me)-βAla-OH: To the crude N-Boc-3-iodo-Tyr(Me)-OH (2.53 g, 6 mmol) in DME (12 niL) cooled at O ⁰ C was added DCC (1.32 g, 6.4 mmol) and SuOH (0.74 g, 6.4 mmol). The mixture was allowed to warm up overnight before filtration of the DCU precipitate. Washing of the solid with AcOEt and evaporation of the filtrate afforded the activated tyrosine ester (3 g, 100%) which was used in the next step without further purification. To a solution of the crude activated ester (200 mg, 0.36 mmol) in DMF was added βAla (35 mg, 0.38 mmol). The mixture was stirred overnight before being-diluted with CH ₂ Cl ₂ and washed with a 10% aqueous KHSO ₄ solution. Drying over Na ₂ SO ₄ and evaporation to dryness gave N-Boc-3-iodo-Tyr(Me)-β AIa-OH. Purification by column chromatography (0-20% MeOH/CH ₂ Cl ₂ ) afforded N-Boc-3-iodo-Tyr(Me)-β AIa-OH (132 mg, 70%) as a white solid. ¹ H NMR (300MHz, CDCl ₃ ) δ 1.3 (s, 9H, (CH ₃ ) ₃ ), 2.36 (m, 2H, CH ₂ ), 2.57-2.65 (m, IH, CH ₂ ), 2.8-2.84 (m, IH, CH ₂ ), 3.23-3.33 (m, 2H, CH ₂ ), 3.78 (s, 3H, OCH ₃ ), 4.02 (m, IH, CH), 6.87 (d, J - 8.7, IH, NHBoc), 6.91 (d, J = 8.4, IH, H5), 7.23 (d, J = 7.2, IH, NH), 7.66 (m, IH, aromatic H), 7.97 (m, IH, aromatic H). Anal. Calcd. for C ₁₈ H ₂₅ TN ₂ O ₆ , H ₂ O: C, 42.36; H, 5.33; N, 5.49. Found: C, 42.03; H, 5.05; N, 5.34.

,44

N-Boc-3-iodo-Tyr(Me)-βAla-7-bromo-Trp-OMe: compound A493. To a suspension of N-Boc-3-iodo-Tyr(Me)-βAla-OH (81 mg, 0.24 mmol), Boc-3-iodo-Tyr(Me)-OH (120 mg,

0.24 mmol), EDC (52 mg, 0.27 mmol) and HOBt (37 mg, 0.27 mmol) in CH ₂ Cl ₂ (3 mL) was added NEt ₃ (75 μL, 0.54 mmol) at 0 ⁰ C. The resulting solution was allowed to warm up to room temperature overnight. The reaction mixture was washed successively with aqueous 5%

KHSO ₄ , aqueous 0.5 M KHCO ₃ and brine. The organic layer was dried over Na ₂ SO ₄ and the solvent was removed in vacuo. The residue was subjected to flash chromatography on silica

gel (1-3% MeOH/CH ₂ Cl ₂ ) to afford N-Boc-3-iodo-Tyr(Me)-βAla-7-bromo-Trρ-OMe (143 mg, 76%) as a white amorphous solid. ¹ H NMR (300MHz, CDCl ₃ ) : δ 1.38 (s, 9H, (CH ₃ ) ₃ ), 2.23-2.45 (m, 2H, CH ₂ ), 2.70-3.04 (m, IH, CH ₂ Tyr), 3.21-3.41 (m, 3H ₅ CH ₂ Trp, CH ₂ ), 3.61- 3.72 (m, IH, CH ₂ ), 3.79 (s, 3H, OCH ₃ ), 3.85 (s, 3H, OCH ₃ ), 4.19-4.32 (m, IH, CHa Tyr), 4.83-4.93 (m, IH, CHa Trp), 5.16 (m, IH, NH), 6.36 (m, IH, NH), 6.74 (d, J = 8.4, H5 Tyr), 6.8 (broad s, IH, NH), 7.03 (t, J = 7.7 Hz, IH, H7 Trp), 7.13 (m, 2 aromatic H), 7.36 (d, J = 7.6 Hz, IH, H4 Trp), 7.49 (d, J = 7.9 Hz, H6 Trp), 7.57 (d, J = 2 Hz, IH, H2 Tyr), 8.52 (broad s, IH, NHind). ¹³ C NMR (75 MHz, CDCl ₃ ) δ 27.3 (CH ₂ Trp), 28.3 ((CH ₃ ) ₃ ), 35.9 (2 CH ₂ ), 37 (CH ₂ Tyr), 52.8 (OCH ₃ ), 36.4 (OCH ₂ ), 80.3 (C(CHs) ₃ ), 85.9 (C3 Tyr), 105 (C7 Trp), 110.9 (CH), 111.4 (C), 117.7(CH), 120.9(CH), 123.4(C), 124.7(CH), 128.6(C), 130.4(CH), 131(C), 134.9(C), 140.2(CH), 156 (C4 Tyr), 157 (CO Boc), 171.6, 171.8 (CO amide and CO ester). Anal. Calcd. for C ₃₀ H ₃₆ IN ₄ O ₇ , 0.5 H ₂ O: C, 46.17; H, 4.78; N, 7.18. Found: C, 46.39; H, 4.83; N, 7.04.

NH ₂ -(Bn)AIa-OMe hydrochloride salt : BocNH-(Bii)Ala (200 mg, 0.68 mmol prepared according to Hannachi, J. C; Vidal, J.; Mulatier, J. C; Collet, A., Electrophilic animation of amino acids with N-Boc-oxaziridines: efficient preparation of N-orthogonally diprotected hydrazino acids and piperazic acid derivatives. J. Org. Chem. 2004, 69, (7), 2367-2373) was solubilized in a 4.5 M anhydrous HCl solution in MeOH. The mixture was allowed to stir overnight and concentrated off. Precipitation of the residue in Et ₂ O and filtration afforded HCl, NH ₂ -(Bn)AIa-OMe (169 mg, 100%) as a white solid. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.38 (d, J ^« 7.2 Hz, 3H, CH ₃ ), 3.41 (s, 3H, OCH ₃ ), 3.95 (q, J = 7.2 Hz, IH, CH), 4.04 (d, J = 13.8 Hz, IH, CH ₂ ), 4.14 (d, J = 13.8 Hz, IH, CH ₂ ), 7.34 (m, 5 aromatic H), 9.71 (broad s, 3H ₅ NH ₃ ⁺ ). 13C NMR (75 MHz, DMSO-d ₆ ) δ 14.3 (CH ₃ ), 52.6 (OCH ₃ ), 57.8 (CH ₂ ), 58 (CH) ₅ 128.5 (CH), 128.7 (CH) ₅ 129.6 (CH) ₅ 134 (C), 172 (CO ester).

N-Boc-3-iodo-Tyr(Me)-NH-(Bn)AIa-OMe: To a suspension of HCl, NH ₂ -(Bn)AIa-OMe (100 mg, 0.41 mmol), Boc-3-iodo-Tyr(Me)-OH (172 mg, 0.41 mmol) and PyBOP (213 mg, 0.41 mmol) HOBt (37 mg, 0.27 mmol) in CH ₂ Cl ₂ (1 mL) was added DIEA (196 μL, 1.13 mmol) at 0 ⁰ C. The resulting solution was allowed to warm up and was stirred for 2 h. The reaction mixture was diluted in AcOEt and washed successively with aqueous 5% KHS 04, aqueous 0.5 M KHCO ₃ and brine. The organic layer was dried over Na ₂ SO ₄ and the solvent was removed in vacuo. The residue was subjected to flash chromatography on silica gel (30% AcOEt/cyclohexane) to afford N-Boc-3-iodo-Tyr(Me)-NH-(Bn) AIa-OMe (165 mg, 66%) as a white amorphous solid. ¹ H NMR (300 MHz, CDCI ₃ ) δ 1.31 (d, J = 7.2 Hz, 3H, CH ₃ ), 1.4 (s, 9H, (CHs) ₃ ), 2.76 (m, 2H, CH ₂ Tyr), 3.62 (q, J = 7.3 Hz, IH, CH Ala), 3.66 (s, 3H, OCH ₃ ), 3.79 (s, 3H, CH ₃ ), 3.86 (m, 2H, CH ₂ Bn), 4.1 (m, IH, CH Tyr), 5.09 (m, IH, NHBoc), 6.65 (d, J = 8.4 Hz, IH, H5), 7.02 (dd, J = 8.4 Hz, J = 2 Hz, IH, H6), 7.22-7.33 (m, 5 aromatic H), 7.55 (d, J = 2 Hz, IH, H2), 7.48 (s, IH, NH). ¹³ C NMR (75 MHz, DMSOd ₆ ) δ 16.3 (CH ₃ ), 28.3 ((CH ₃ ) ₃ ), 36.9 (CH ₂ Tyr), 51.7 (OCH ₃ ), 54.8 (CH Tyr), 56.4 (OCH ₃ ), 59.9 (CH Ala), 60.2 (CH ₂ Bn), 80.1 (C(CH ₃ ) ₃ ), 86 (C3), 110.9 (CH), 127.7 (CH), 128.3 (CH), 129.2 (CH), 130.4 (CH), 130.8 (C), 136.2 (C), 140.1 (CH), 155.2 (C4), 157.1 (CO Boc), 169.7 (CO amide), 174.4 (CO ester).

N-Boc-3-iodo-Tyr(Me)-NH-(Bn)AIa-7-bromo-Trp-OMe. To a solution of N-Boc-3-iodo- Tyr(Me)-NH-(Bn)Ala-OMe (135 mg, 0.22 mmol) in THF (3 mL) cooled at 0 °C was added a 1 M aqueous LiOH solution (0.25 mL). The mixture was stirred for 1 h 30 at 0 °C before it was quenched by 2 N aqueous HCl solution (pH = 1-2). The aqueous phase was extracted twice by CH ₂ Cl ₂ , the combined organic layers were dried over Na ₂ SO ₄ . Removing of the solvents in vacuo afforded the crude N-Boc-3-iodo-Tyr(Me)-NH-(Bn) AIa-OH (121 mg, 92%) as a white foam which was used in the next step without further purification. To a suspension of 7-bromo-Trp-OMe (56 mg, 0.17 mmol), N-Boc-3-iodo-Tyr(Me)-NH-(Bn)Ala-OH (100 mg, 0.17 mmol), EDC (35 mg, 0.18 mmol) and HOBt (25 mg, 0.18 mmol) in CH ₂ Cl ₂ (2 mL) was added NEt ₃ (52 μL, 1.13 mmol) at 0 °C. The resulting solution, allowed to warm up was stirred 2 h. The reaction mixture was diluted in AcOEt and washed successively with aqueous 5% KHSO ₄ , aqueous 0.5 M KHCO ₃ and brine. The organic layer was dried over Na ₂ SO ₄ and

the solvent was removed in vacuo. The residue was subjected to flash chromatography on silica gel (5% MeOH/CH ₂ Cl ₂ ) and precipitated in AcOEt/pentane to afford N-Boc-3-iodo- Tyr(Me)-NH-(Bn)Ala-7-bromo-Trρ-OMe (77 mg, 52%) as a white amorphous solid. ¹ H NMR (300 MHz ₅ CDCl ₃ ) δ 1.17 (d, J = 7 Hz, 3H, CH ₃ ), 1.39 (s, 9H, (CH ₃ ) ₃ ), 2.49-2.69 (m, 2H, CH ₂ Tyr), 3.22-3.32 (m, 2H, CH ₂ Trp), 3.41-3.51 (m, IH, CH Ala), 3.67 (s, 3H ₅ OCH ₃ ), 3.7-3.96 (m, 2H, CH ₂ Bn), 3.8 (s, 3H, OCH ₃ ), 3.97-4.08 (m, IH, CH Tyr), 4.82-4.93 (m, IH, CH Trp), 5.04 (d, J = 8.2 Hz ₅ IH, NHBoc), 6.62 (d, J = 8.2 Hz, IH, H5 Tyr), 6.93 (dd, J = 8.2 Hz ₅ J - 2 Hz, IH, H6 Tyr), 6.99 (t, J = 7.7 Hz, IH, H5 Trp), 7.19-7.24 (m, 6 aromatic H), 7.32 (d, J = 7.5, 1 aromatic H), 7.5 (d, J = 2 Hz ₅ IH, H2 Tyr), 7.56 (d, J = 7.9 Hz, 1 aromatic H), 7.71 (s, IH, NH), 8.08 (s, IH, NH), 8.65 (s, IH, NH). ¹³ C NMR (75 MHz, DMSO-d ₆ ) δ 13.4 (CH ₃ ), 27.6 (CH ₂ Trp), 28.3 ((CHs) ₃ ), 36.1 (CH ₂ Tyr), 52.5 (OCH ₃ ), 54.8 (CH Tyr), 56.4 (OCH ₃ ), 59.9 (CH ₂ Bn), 62.6 (CH Tyr, CH Ala), 80.4 (C(CH ₃ ) ₃ ), 86 (C3), 104.8 (C7 Trp), 110.9 (CH), 111.7 (C), 118 (CH), 120.6 (CH), 123.8 (CH), 124.5 (CH), 127.8 (CH), 128.4 (CH), 128.7 (C), 129.2 (CH), 130.3 (CH) ₅ 130.4 (C), 130.9 (CH), 131 (C), 134.8 (C), 135.7 (CH), 139.9 (CH) ₅ 155.6 (C4), 157 (CO Boc), 170.6, 172.6, 172.7 (2 CO amide, CO ester). Anal. Calcd. for C ₃₇ H ₄₅ N ₅ O ₇ BrI, H ₂ O: C, 49.56; H ₅ 5.28; N, 7.81. Found: C ₅ 49.83; H ₅ 5.08; N, 7.57.

N-Boc-Tyr(Me)-NH-(Bn)Ala-Trp-OMe: Treatment of N-Boc-3-iodo-Tyr(Me)-NH-(Bn) Ala-7-bromo-Trp-OMe in the condition of the suzuki coupling according to Berthelot, A.; Piguel, S.; Le Dour, G.; Vidal, J., Synthesis of macrocyclic peptide analogues of proteasome inhibitor TMC-95A. J. Org. Chem. 2003, 68, (25), 9835-9838 afforded N-Boc-Tyr(Me)- NH(Bn)Ala-Trp-OMe. HRMS (ESI) calcd for C ₃₇ H ₄₅ N ₅ O ₇ Na [(M+Na) ⁺ ] 694.3217, found 694.3197.

N-Boc-Tyr(Me)-βAIa-Trp-OMe: Treatment of N-Boc-3-iodo-Tyr(Me)-βAla-7-bromo- Trp-OMe in the condition of the suzuki coupling according to Berthelot, A.; Piguel, S.; Le Dour, G.; Vidal, J., Synthesis of macrocyclic peptide analogues of proteasome inhibitor TMC-95A. J. Org. Chem. 2003, 68, (25), 9835-9838 afforded N-Boc-Tyr(Me)-βAla-Trp- OMe. HRMS (ESI) calcd for C ₃₀ H ₃₈ N ₄ O ₇ Na [(M+Na) ⁺ ] 589.2638, found 589.2634.

VIII) Enzymatic evaluation:

Proteasome:

2OS proteasome from rabbit reticulocyte was from commercial source (Alexis Biochemicals)

Enzymatic analysis of inhibition

Semi-automatic fluorescent assays using Suc-LLVY-amc for chymotrypsin-like activity, Z- LLE-BNA for post-acid activity and Boc-LLR-amc for trypsin-like activity of proteasome were performed at pH 7.5 and 30 ⁰ C using BMG Fluostar microplate reader (Sue = succinyl; amc = 7-amino-4-methylcournarin ; Z = benzyloxycarbonyl; NA = 2-naphtylamine; Boc = tert-butoxycarbonyl). The buffers were: 20 mM Tris, 1 mM DTT, 10 % glycerol, 3 % (v/v) DMSO (ChT-L and PA activities) ; 20 mM Tris, 1 mM DTT, 10 % glycerol, 3 % (v/v) DMSO. Studied compounds were dissolved in DMSO prior dissolution in the buffer. Proteasome was incubated for 15 min at 30 ⁰ C in the presence of the studied compound (0.1- 100 μM). A control assay in the absence of tested compounds contained DMSO at the same concentration (3 %, v/v). The fluorogenic proteasome substrate was then added and the hydrolysis of the appropriate fluorescent substrate was monitored for 1 h (λ _exc = 360, λ _em = 465 nm for amc substrates and λ _exc = 340 , λ _em = 405 nm for the BNA substrate). Initial rates determined in control experiments were considered to be 100 % of the peptidasic activity; initial rates that were above 100 % in the presence of a test compound were considered to be activations, while initial rates below 100 % were considered to be inhibitions. Por weak inhibitors, the percentage of inhibition at a reference concentration (100 μM) is reported. The results, expressed in % inhibition (or activation factor), were obtained by calculating the

average of at least two independent experiments, the variability was less than 10 %. The inhibitory activity of more efficient compounds are expressed as IC ₅₀ calculated by fitting the experimental data to the equation 1: % Inhibition = 100 [I] ₀ /(IC ₅₀ + [I] ₀ ) = 100 (v ₀ - V;/ V ₀ ), or equation 2 : % Inhibition = 100 [I] ₀ ^nH /(IC ₅₀ ^nH + [I] ₀ ^nH ), nH is the Hill number. V ₀ and v; are the initial rates in the absence and in the presence of inhibitor. A Dixon plot has been used to determine the inhibition constant Ki for competitive inhibition by compound A215. The reversible character of inhibition or activation was determined by measuring the activity of treated enzyme after the inhibitor molecule has been withdrawn from the reaction medium.

Results:

Inhibition of rabbit 20 S proteasome (pH = 7.5, 30 ⁰ C)

ni : non inhibitor

CT-L PA T-L

% inhibition (at 100 μM) % inhibition (at lOO μM) % inhibition (at 100 μM) or IC ₅₀ or IC ₅₀ or IC ₅₀

Compounds Il

A374F1 ni 18%

A291 ni 19%

A389F1pl2 ni 55% ni

Compounds III

SP221 ni ni

SP225F2 ni 25% 49%

SP226F1 ni ni 23%

Compounds IV-1A

A248 ni 32% ni

A215 IC ₅₀ = 6.8 μM (Ki = 2 μM) IC ₅₀ = 11.3 μM IC ₅₀ = 14.4 μM

SP274 62% 45% ni

A363 ni 20%

A340 ni 17% ni

A174 ni ni ni

A268 ni ni

A385 28% 38% ni

A254 38% 60% ni

Compounds IV-2

PSV11 R 59%

NR35 ni ni

SP303r2 ni

SP304R ni 45%

SP313P 26% IC ₅₀ = 4 μM ni

NR36 34% 39%

NR40 IC ₅₀ = 40 μ M IC ₅₀ = 35 μM

A424P ni

A414P ni

A418P ni 32%

SP296P 22% 59%

SP314C2 ni

A416 28% 35%

SP318C 15% 66%

SP325 IC ₅₀ = 5.4 μM IC ₅₀ = 2.5 μM IC ₅₀ = 19 μM

SP324 IC ₅₀ = 9 μM IC ₅₀ = 3 μM IC ₅₀ = 21 μM

SP310C ni

SP315C2 . ni

SP320P2 ni ni ni

SP306P ni

SP307P ni

SP319P 36% ni 14%

SP308P ni

Compounds IV-3

CV11 ni ni

CV12 ni IC ₅₀ = 10,4 μM

CV13 32% IC ₅₀ = 3,9 μM ni

JV602 22% 26%

NR15 IC ₅₀ = 2.2 μM 87%

NR38 IC ₅₀ = 13.5 μM 81%

NR16 12% ni

Compound IV-1 B

A493 29% 40%

Activation of rabbit 20 S proteasome (pH = 7.5, 30 ⁰ C, [compound] = 100 μM)

CT-L PA T-L Activation factor Activation factor Activation factor

Compounds Il

A374F1 1.8 A291 1.4

Compounds III

SP221 1.4

Compounds IV-1A

A363 1.9 A268 2

Compounds IV-2

PSV11 R 1.2 1.7

NR35 1.8

SP303r2 1.4 ' 1.6

SP304R 1.7

SP305R 1.5 1.2 3.2

NR36 9.8

NR40 6

A424P 1.2 1.9

A414P 1.3 1.9

A418P 1.6

SP296P 2.3

SP314C2 1.5 2.5

A416 2.2

SP318C 1.2

SP323C2 1.6 1.2 2.2

SP310C 1.7 1.6

SP315C2 1.2 1.9

SP311C 1.3 1.2 1.9

SP306P 1.2 1.6

SP307P 1.3 1.8

SP308P 1.6 1.9

Compounds IV-3

CV11 1 ,7

CV12 1 ,5

JV602 1 ,3

NR15 12.3

NR38 9.8 NR16 8^

Compound IV-1B

A493 8.2

Figure legends

Figure 1: Schematic representation of the reaction of: (A) peptidic inhibitors

(aldehydes, boronates, vinylsulfones), and (B) non peptidic inhibitors such as clasto- lactacystin-β-lactone, (-)epigallocatechin-3-gallate with the catalytic Thrl of the active sites of proteasome. Adducts (A) or stable acyl-enzymes (B) are obtained after the formation of a covalent bond between Thrl and the reactive group of the inhibitor.

Figure 2 A: Mechanism of inhibition of proteasome by Velcade

Figure 2B: Structures of proteasome inhibitors

Figure 3: Structures of known non covalent inhibitors of proteasome