MXPA00010056A

MXPA00010056A - Process for the preparation of resin-bound cyclic peptides

Info

Publication number: MXPA00010056A
Application number: MXPA/A/2000/010056A
Authority: MX
Inventors: James J Mencel; Adam W Sledeski
Original assignee: James J Mencel; Rhonepoulenc Rorer Pharmaceuticals Inc; Adam W Sledeski
Priority date: 1998-04-15
Filing date: 2000-10-13
Publication date: 2001-07-31

Abstract

This invention is directed to a process for the solid phase, fragment-based synthesis of resin-bound cyclic peptide analogs of parathyroid hormones and analogs of parathyroid hormone-related proteins, which analogs contain at least one bridge between the side chains of two non-adjacent amino acid residues, and to peptide fragments useful therefor.

Description

ROCEDURE FOR THE PREPARATION OF CYCLIC PEPTIDES UNITED TO RESIN FIELD OF THE INVENTION This invention relates to a process for the preparation based on fragments of cyclic peptide analogues bound to parathyroid hormone resin and parathyroid hormone-related protein analogs, whose analogues contain at least one bridge between the side chains of two amino acid residues not adjacent More particularly, the invention relates to a process for the solid phase synthesis of said cyclic peptide analogs and for peptide fragments useful therefor.

BACKGROUND OF THE INVENTION The cyclic peptide subunits are present in a wide variety of peptides possessing useful biological activity, including parathyroid hormone analogues and protein analogs related to parathyroid hormone, vasoactive peptide analogs, cholecystokinin analogs, peptides derived from tumor necrosis factor ( TNF), calcitonin analogues, somatostatin analogues, cell adhesion modulators, hormone release factor analogues and growth (GRF), bradykinin antagonists, analogs of tyrosine activation motifs (TAM mimetics) and amylin agonists. Of particular interest are the cyclic peptide parathroid hormone (hPTH) analogs and protein analogs related to parathyroid hormone (hPTHrP). Human parathyroid hormone (hPTH) is a protein of 84 amino acids, which is a major regulator of calcium homeostasis. The protein related to parathyroid hormone hPTHrP, is a protein of 139 to 171 amino acids with N-termlnal homology with hPTH. The fragments N-terminal hPTH and hPTHrP, particularly those that consist of amino acids 1-34, retain all the biological activity of the hormone of origin. hPTH (1-34) has the following amino acid sequence: Ser-Val-Ser-Glu-lle-GIn-Leu-Met-His-Asn-Leu-Gly-Lys-His-Leu-Asn-Ser-Met-Glu -Arg-Val-Glu-Trp-Leu-Arg-Lys-Lys-Leu-Gln-Asp-Val-His-Asn-Phe. hPTHrP (1-34) has the following amino acid sequence: Ala-Val-Ser-Glu-His-GIn-Leu-Leu-His-Asp-Lys-Gly-Lys-Ser-lle-Gln-Asp-Leu-Arg -Arg-Arg-Phe-Phe-Leu-His-Hls-Leu-lle-Ala-Glu-lle-His-Thr-Ala. The biological activity of hPTH is reflected in the activation of two secondary messenger systems: adenylyl cyclase activity (AC) coupled to protein G and protein kinase C (PKC) activity not coupled and coupled to protein G. N-terminal fragments hPTH (1-34) 0H and hPTH (1-31) NH2 have proved to be anabolic with respect to bone formation in humans and ovariectomized rats, respectively. It has been shown that this increase in bone growth is coupled with stimulation of adenylyl cyclase activity. The analogues of these N-terminal fragments have an important therapeutic potential for the treatment of physiological conditions associated with bone cell calcium regulation including hypocalcemia; osteoporosis; osteopenia and disorders associated with osteoporosis and osteopenia such as hyperparathyroidism, hypoparathyroidism and Cushing's syndrome; osteopenia induced by glucocorticoid and immunosuppressant; and repair of bone fracture and bone fracture. It has also been established that the deletion of up to six amino acid residues from the N-terminus of hPTH (1-34) markedly decreases the ability of the resulting analog to stimulate adenylyl cyclase, while having a small effect on receptor binding. Thus, hPTH analogues (1-34) truncated by up to six amino acid residues at the N-terminus inhibit the action of PTH and are useful in the treatment of disorders characterized by an excess of PTH, such as hyperparathyroidism and crisis. of hypercalcemia related to hyperparathyroidism, hypercalcemia of malignancy, renal failure and hypertension. The acyclic analogues of hPTH (1-27) to (1-34) are described in the patent of E.U.A. 4,086,196. The acyclic analogs of hPTH (1-34) and hPTHrP (1-34) are described in the U.S. patent. No. 5,589,452. [Nle8, Nle18, Tyr34, or Phe34] hPTH (1-34) are described in the patent of E.U.A. Do not. 4,656,250. [Nle8, Nle18, Tyr3] hPTH (1-34) and N-truncated derivatives thereof are described in the U.S. Patents. Nos. 4,771, 124 and 4,423,037. Other acyclic analogues of PTH (1-34) are described in the patents of E.U.A. Nos. 5,723,577 and 5,434,246, WO 97/02834, EPA 561 412-A1, EPA 747 817-A2, WO-94/02510, WO9603437, and W09511988-A1. Analogs of hPTH (1-28) NH2 to hPTH (1-31) NH2 and [Leu27] hPTH (1-28) NH2 to [Leu27] hPTH (1-33) NH2 are described in the patent of E.U.A. No. 5,556,940. Acyclic PTH receptor antagonists that include N-terminally truncated PTH analogs are described in the U.S. Patents. Nos. 5,446,130, 5,522,489, 4,771, 124 and 4,423,037. The cyclic and bicyclic analogs of hPTH and hPTHrP have been described. Cyclo (Lys26-Asp30) [Leu27] hPTH (1-34) NH2 and cyclo (Lys27-Asp30) hPTH (1-34) NH2 are described in the patent of E.U.A. No. 5,556,940. Cycle (Lys26-Asp30) [Leu27] hPTH (1-31) NH2, Cyclo (Glu22-Lys26) [Leu27] hPTH (1-31) NH2, and Cyclo (Lys27-Asp30) hPTH (1-31) NH2 are described by Barbier, et al., J. Med. Chem. 1997, 40.1373. The monocyclic and bicyclic derivatives of hPTH (1-34) or hPTHrP (1-34) are described in WO 96/40193, DE19508672-A1, and by A. Bisello, et al., In Biochemistry 1997, 36 , 3293. Cycle (Lys13-Asp17) hPTHrP (7-34) NH2, a potent PTH receptor antagonist, is described by M. Chorev, et al., Biochemistry 1991, 30, 5698. In addition, Kanmera, et al., has described a series of hPTHrP amide-containing analogues, Peptide Chemistry 1993: Okada, Y., ed .; Protein Research Foundation, Osaka, 1994, 321-324. " WO 98/51324 discloses cyclic peptide compounds of formula I X-Al0-All-A12-A13-Al4-Al5-Al6-A17-Al8-A19-A20-A2l-A22-A23-A24-A25-26-A27- YI and pharmaceutically acceptable salts and prodrugs thereof, wherein X is selected from the group consisting of (a) R? A-Ao-A A2-A3-A4-A5-A6-A7-A8-A9-, (b ) R a-A2-A3-A4-A5-A6-A7-A8-A9-, (c) R | b-A3-A4-A5-A6-A7-A8-A9-, (d) R | a -A4-A5-A6-A7-A8-Ag-, (e) ia-A5-A6-A7-A8-A9-, (f) Rla-A6-A7-A8-A9-, (I) R? A-Asr, and And it is selected from the group consisting of (a) -Rs. (c) -A28-A2 -R3, (d) -A28-A29-A3o-R3, (e) -A28-A29-A3o-A31-R3, (f) -A28-A29-A3o-A3i-A32-R3, (g) -A28-A29-A3o-A3? -A32-A33-R3, and (h) A28-A29-A30-A31-A32-A33 -A3 -R3; R a is H, alkyl, araiquyl or -COR2; Rib is R? A or a group of formula 2 is alkyl, alkenyl, alkynyl, aryl or aralkyl; R3 is a group of formula A35-OR4 or A35-NR4R5; R and R5 are independently H or lower alkyl; Re and Rg are independently H or alkyl; R7 is alkyl; R8 is H, alkyl or COR2; R10 is H or halogen; R11 is alkyl or aralkyl; m is 1, 2 6 3; n is 3 or 4; A0 is absent or is a peptide of one to six amino acid residues; A, is Ser, Ala, Gly or D-Pro, or an equivalent amino acid thereof; A2 is Ala, Val or Gly, or an equivalent amino acid thereof; A3 is Ala, Ser, Gly or D-Pro, or an equivalent amino acid thereof; A4 is Glu, ala or Gly, or an equivalent amino acid thereof; A5 is Lie, His, Ala or Gly, or an equivalent amino acid thereof; e is Ala, Gln, Gly or D-Pro, or an equivalent amino acid thereof; A7 is Ala, Leu, Gly, or an equivalent amino acid thereof; Ace is Leu, Nle, Gly or D-Pro, or an equivalent amino acid thereof; A9 is His, Ala, D-Pro or Gly, or an equivalent amino acid thereof; A10 is Ala, Asn, Asp, Cys, homo-Cys, Glu, Gly, Lys, Orn, Ser, Thr, D-Pro, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO-; A11 is Ala, Gly, Leu or Lys, or an equivalent amino acid thereof; A-? 2 is Ala or Gly, or an equivalent amino acid thereof; A13 is Ala, Asn, Asp, Cys, homo-Cys, Glu, Gly, Lys, Orn, Ser, Thr, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO-; A is Ala, Asn, Asp, Cys, homo-Cys, Glu, Gly, Lys, Orn, Ser, Thr, D-Pro, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO-; Ais is Ala, Gly, Lie, D-Pro or Leu, or an equivalent amino acid thereof; A16 is Asn, Ala, Gly, D-Pro or Gln, or an equivalent amino acid thereof; Ai? is Ala, Asn, Asp, Cys, homo-Cys, Glu, Gly, Lys, Orn, Ser, Thr, D-Pro, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO-; A18 is Ala, Asn, Asp, Cys, homo-Cys, Glu, His, Leu, Lys, Orn, Nle, Ser, Thr, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO-; A19 is Arg or Glu, or an equivalent amino acid thereof; A2o is Arg, or an equivalent amino acid thereof; A2? is Arg, Asp, Cys, homo-Cys, Glu, Lys, Orn, Ser, Thr, Val, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO-; A22 is Asp, Cys, homo-Cys, Glu, His, Lys, Orn, Phe, Ser, Thr, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO-; A23 is Leu, Phe or Trp, or an equivalent amino acid thereof; A24 is Leu, or an equivalent amino acid thereof; A25 is Arg, Asp, Cys, homo-Cys, Glu, His, Lys, Orn, D-Pro, Ser, Thr, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO; A2ß is Asp, Cys, homo-Cys, Glu, His, Lys, Orn, Ser, Thr, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO; A27 is Leu or Lys, or an equivalent amino acid thereof; A28 is either Leu, or an equivalent amino acid thereof; A29 is Ala, Asp, Cys, homo-Cys, Glu, Gln, Lys, Orn, Ser, Thr, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO; A3o is Asp, Cys, homo-Cys, Glu, Gly, Lys, Orn, Ser, Thr, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO; A3? is Lie, Leu or Val, or an equivalent amino acid thereof; A32 is His, or an equivalent amino acid thereof; A33 is Asn or Thr or an equivalent amino acid thereof; A34 is Ala or Phe, or an equivalent amino acid thereof; A35 is absent or is a peptide of 1 to 4 amino acids; and the side chains of at least one of the following pairs of amino acid residues, A10 and Au, A13 and A17, Au and A-ie, A17 and A21, A-is and A22, A2? and A2, A25 and A2, A26 and A3o, are linked through an amide, ester, disulfide or lanthionine bond to form a bridge, and the side chain of each of the following amino acid residues A10, A13, Au, A17, A18, A2 ?, A22, A25, A26, A29, and A30, contributes largely to the formation of a single bridge; since when the side chains of the following pairs of acidic amino acid residues, Aβ3 and A17 or A2β and A3o are linked through an amide, disulfide or lanthionine bond to form a bridge, then the side chains of at least one of the following pairs of amino acid residues, A10 and Au, A14 and A? 8, A17 and A2 ?, A18 and A22, A2? and A25, A25 and A2g are also linked through an amide, ester, disulfide or lanthionine linkage.

A preferred subset of the peptide compounds of formula I comprises the peptide compounds, wherein X is selected from the group consisting of (a) R ^ -Ao-A ^ As-A ^ -As-Ae-Ar-Aβ- Ag-, (b) R1a-A2-A3-A4-A5-A6-A7-A8-Ag-, (c) R1b-A3-A4-A5-A6-A7-A8-A9-, (d) Ria- A4-A5-A6-A7-A8-A9-, (e) R1a-A5-A6-A7-A8-Ag-, (0 Ria-A6-A7-A8-Ag-, (g) R1a-A7-A8 -A9-, (h) Ria-A8-A9-, (i) R1a-A9-, and Y is selected from the group consisting of (a) -R3, (b) -A28-R3, (c) -A28-A29-R3, (d) -A28-A29-A3o-R3, (e) -A28- A29-A3o-A31-R3, (f) -A28-A29-A3o-A3? -A32 -R3, (g) -A28? 29-A3o-A3i-A32-A33-R3, and (h) -A28- 29-A30-A31-A32-A33-A34-R3; R1a is H, alkyl, aralkyl or -COR2; R-ib is R-ia or a group of formula R2 is alkyl, alkenyl, alkynyl, aryl or aralkyl; R3 is a group of formula A35-OR or A35-NR4R5; R4 and R5 are independently H or lower alkyl; R6 and Rg are independently H or alkyl; R7 is alkyl; R8 is H, alkyl or COR2; R10 is H or halogen; R11 is alkyl or aralkyl; Ao is absent or is a peptide of one to six amino acid residues; Ai is Ser, Ala, Gly or D-Pro, or an amino acid equivalent of the A2 is Ala, Val or Gly, or an equivalent amino acid thereof; A3 is Ala, Ser, Gly or D-Pro, or an amino acid equivalent of the A4 is Glu, Ala or Gly, or an equivalent amino acid thereof; As is lie, His, Ala or Gly, or an equivalent amino acid thereof; Aβ is Ala, Gln, Gly or D-Pro, or an equivalent amino acid thereof; A7 is Ala, Leu or Gly, or an equivalent amino acid thereof; A8 is Leu, Nle, Gly or D-Pro, or an amino acid equivalent thereof; Ag is His, Ala, Gly or D-Pro, or an equivalent amino acid thereof; A-io is Ala, Asn, Gly, Lys, Asp or D-Pro, or an equivalent amino acid thereof; An is Ala, Gly, Lys, Leu or Lys, or an equivalent amino acid thereof; A12 is Ala or Gly, or an equivalent amino acid thereof; A? 3 is Ala, Gly or Lys, or an equivalent amino acid thereof; A is Ala, Gly, His, Ser, Asp, Lys or D-Pro, or an equivalent amino acid thereof; Ais is Ala, Gly, Lie, D-Pro or Leu, or an equivalent amino acid thereof; Ate is Asn, Ala, Gly, D-Pro or Gln, or an equivalent amino acid thereof; Ai? is Ala, Asp, Gly, Ser, Lys or D-Pro, or an equivalent amino acid thereof; Ais is Lys, or an equivalent amino acid thereof; A-ig is Arg or Glu, or an equivalent amino acid thereof; A2o is Arg, or an equivalent amino acid thereof; A2? is Arg, Lys, Asp, or Val, or an equivalent amino acid thereof; A22 is Asp, Lys, Orn, or Glu, or an equivalent amino acid thereof; A23 is Leu, Phe or Trp, or an equivalent amino acid thereof; A24 is Leu, or an equivalent amino acid thereof; A25 is Arg, His, Asp, Lys or Glu, or an equivalent amino acid thereof; A 6 is Lys or His, or an equivalent amino acid thereof; A27 is Leu or Lys, or an equivalent amino acid thereof; A28 is either Leu, or an equivalent amino acid thereof; A29 is Ala, Asp, Glu or Gln, or an equivalent amino acid thereof; A30 is Asp, Lys or Glu, or an equivalent amino acid thereof; A31 is Lie, Leu or Val, or an equivalent amino acid thereof; -. ,-TO" A32 is His, or an equivalent amino acid thereof; A33 is Asn or Thr, or an equivalent amino acid thereof; AJÍ is Ala or Phe, or an equivalent amino acid thereof; A35 is absent or is a peptide of 1 to 4 amino acids. The above peptide compounds are described in WO 98/51324 as possessing useful properties, in particular, pharmaceutical properties. They are especially useful for treating disease states capable of being modulated by compounds which bind to parathyroid hormone receptors either with or without concomitant stimulation of adenylyl cyclase activity. The present invention relates to an improved method for synthesizing these peptide compounds. Solid phase synthesis of cyclic peptides typically involves the consecutive addition of amino acids to a peptide synthesis resin to obtain a resin-bound peptide that possesses all or a portion of the amino acid sequence of the desired cyclic peptide. The side chain residues of amino acids that will be cyclized are then unprotected and cyclization is effected. If the cyclization is carried out before the completion of the entire amino acid sequence, the remaining amino acids are added and the completed peptide is then cleaved from the resin and purified. However, the linear method for the preparation of peptides and for cyclic peptides in general, is often inefficient and therefore, may not be cost-effective for the preparation of large amounts of peptide.

In cases where a peptide is assembled in a linear manner, the ease of purification of the peptide decreases as the number of amino acid residues in the peptide increases. In addition, for cyclic peptides, the preparation of side chain bridge (s) near the end of the synthesis, imparts the problems associated with the preparation of the side chain bridge, including low yield, secondary reactions and removal of impurities, to all the peptide. Consequently, the incorporation of a cyclic unit in a large peptide increases the difficulties inherent in the synthesis of linear peptides. A commercially feasible synthesis of cyclic analogs of hPTH such as those described in WO 98/51324 and discussed above, require a synthetic method which overcomes the complicating factors of extensive and cyclic components and which allows the preparation of commercially useful amounts in the drug manufacturing context. The present invention relates to a more efficient preparation of cyclic analogues of hPTH and hPTHrP through a process using bound and unbound fragments.

BRIEF DESCRIPTION OF THE INVENTION This invention relates to a method for preparing cyclic analogues of hPTH and hPTHrP of formula II where J, L, and M are fragments of linear peptides K? is absent or is a cyclic peptide fragment, and K2 is a cyclic peptide fragment; this method consists of the steps of: (1) preparing M M - ((RR)) by consecutive addition of amino acid residues suitably protected to a resin to provide: wherein f R) is a suitable peptide synthesis resin and M is a fragment of polypeptide (2) prepared separately by conventional peptide synthesis, a protected N-terminal cyclic polypeptide fragment of formula IV PHMN .C H K rv wherein P is a suitable amine protecting group, (3) coupling III with IV to provide a peptide of formula V (4) if K2 is absent, then (a) the polypeptide fragments J and L are prepared as a single polypeptide of formula VI and polypeptide VI is coupled to peptide V to provide a peptide of formula VII or optionally, (b) the individual amino acid elements protected from J and L polypeptides are added sequentially to the peptide fragment of formula V, '10 or optionally, (c) either or both of J and L are prepared separately as fragments of polypeptide and are coupled to the growing peptide starting with the fragment of formula V, (5) when the cyclic peptide Ki is then present (a) a polypeptide fragment of formula VIII VIII wherein P is a suitable amine protecting group, is prepared separately by means of Conventional peptide synthesis, (b) a peptide fragment of formula IX PH ^ ^ C02H IX is prepared and coupled to the peptide V fragment to provide a peptide fragment of formula X and (c) the cyclic peptide fragment VIII is coupled to the peptide fragment of formula X to provide a peptide fragment of formula XI PHN ^ / ^ f ^ R) ?? (d) the peptide fragment of formula XII is prepared PHN. ^ CQzH XII and fragment XII is coupled to fragment XI, and (6) cleaving the resin and deprotecting. In the process described herein, the cyclic peptide fragments are prepared separately to be coupled with a resin or resin-bound peptide. The separate preparation of the cyclic peptide fragment allows for the convergent synthesis of resin bound cyclic peptides, resulting in increased yield and product of the resulting cyclic peptide. The difficulties associated with the preparation of the linked fragment are limited to a smaller peptide subunit, which can be purified before coupling with the solid phase synthesis resin. The Practice of this technique in the context of certain preferred analogs of hPTH and hPTHrP requires the synthesis of new peptide fragments and new sequences of events.

DETAILED DESCRIPTION OF THE INVENTION As used above and throughout the specification, it should be understood that the following terms have the following meanings, unless otherwise indicated.

Definitions of terms "Alkyl" means an aliphatic hydrocarbon group which may be straight or branched having from about 1 to about 20 carbon atoms in the chain. Branched means that one or more lower alkyl groups are attached to a linear alkyl chain. "Lower alkyl" means about 1 to 4 carbon atoms in the chain which may be straight or branched. Alkyl groups are exemplified by methyl, ethyl, n-e / so-propyl, n-, sec-, iso- and ferbutyl and the like. "Alkenyl" means an aliphatic hydrocarbon group containing a carbon-carbon double bond and which may be straight or branched having from about 2 to about 20 carbon atoms in the chain.

"Lower alkenyl" means from about 2 to about 4 carbon atoms in the chain, which may be straight or branched. The examples of alkenyl groups include ethenyl, propenyl, n-butenyl, / -butenyl, 3-methylbut-2-enyl, p-pentenyl, heptenyl, octenyl, cyclohexylbutenyl and decenyl. f "Alkynyl" means an aliphatic hydrocarbon group containing a carbon-carbon triple bond and which can be straight or branched, having about 2 to about 20 carbon atoms in the chain. "Lower alkynyl" means approximately 2 to 4 carbon atoms in the chain, which can be straight or branched. Examples of alkynyl groups include ethynyl, propynyl, p-butynyl, 3-methybut-2-ynyl, n-f-pentanyl, heptynyl, octynyl and decynyl. "Alkylene" means a divalent group derived from a straight or branched chain saturated hydrocarbon by the removal of two hydrogen atoms, for example methylene, 1,2-ethylene, 1,1-ethylene, 1,3-propylene, 2, 2-dimethylpropylene and the like. "Phenylalkyl" means a phenyl group attached to the molecular moiety of origin via an alkylene group. The alkylene group, preferably, is from about 1 to about 7 carbon atoms. Representative phenylalkyl groups include benzyl, 2-phenylethyl, 2-propylphenyl and the like. "Amine protecting group" means an easily removable group, which is known in the art to protect an amino group against an undesirable reaction during synthetic procedures and for being selectively removable. The use of N-protecting groups is known in the technique for protecting groups against undesirable reactions during a synthetic process and many of these protecting groups are known, compare for example, T.H. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 2nd edition, John Wiley & Sons. New York (1991), incorporated herein by reference. Preferred N-protecting groups are acyl, including formyl, acetyl, chloroacetyl, trichloroacetyl, o-nitrophenylacetyl, o-nitrophenoxyacetyl, trifluoroacetyl, acetoacetyl, 4-chlorobutyryl, isobutyryl, o-nitrocinnamoyl, picolinoyl, acyl isothiocyanate, aminocaproyl, benzoyl and the like, and acyloxy including methoxycarbonyl, 9-fluorenylmethoxycarbonyl, 2,2,2-trifluoroethoxycarbonyl, 2-trimethylsilylethoxycarbonyl, vinyloxycarbonyl, allyloxycarbonyl, 1-butyloxycarbonyl (BOC), 1,1-dimethylpropynyloxycarbonyl, benzyloxycarbonyl (CBZ), p-nitrophenylsulfinyl, p-nitrobenzyloxycarbonyl , 2,4-dichlorobenzyloxycarbonyl, alioxycarbonyl (Alloc), and the like. "Amino acid" means an amino acid selected from the group consisting of natural and non-natural amino acids as defined herein. The amino acids can be neutral, positive or negative depending on the substituents in the side chain. "Neutral amino acid" means an amino acid that contains uncharged side chain substituents. Examples of neutral amino acids include alanine, valine, leucine, soleucine, proline, phenylalanine, tryptophan, methionine, glycine, serine, threonine and cysteine. "Positive amino acid" means an amino acid in which the side chain substituents are positively charged at physiological pH.

Examples of positive amino acids include lysine, arginine, histidine. "Negative amino acid" means an amino acid in which the side chain substituents have a net negative charge at physiological pH. Examples f of negative amino acids include aspartic acid and glutamic acid. The preferred amino acids are amino acids a. The most preferred amino acids are amino acids a which have stereochemistry L on carbon a. "Amino acid residue" means the individual amino acid units incorporated into a peptide or peptide fragment. "Natural amino acid" means an amino acid selected from the group consisting of alanine, valine, leucine, isoleucine, proline, phenylalanine, tryptophan, methionine, glycine, serine, threonine, cysteine, tyrosine, asparagine, glutamine, lysine, arginine, histidine. , aspartic acid and glutamic acid. "Non-natural amino acid" means an amino acid for which there is no nucleic acid codon. Examples of non-natural amino acids include, for example, the D isomers of the natural amino acids as indicated above; Aib (aminobutyric acid), bAib (3-aminoisobutyric acid), Nva (norvaiin), β-Ala, Aad (2-aminoadipic acid), bAad (3-aminoadipic acid), Abu (2-aminobutyric acid), Gaba (acid) α-aminobutyric acid), Acp (6-aminocaproic acid), Dbu (2,4-diaminobutyric acid), α-aminopimelic acid, TMSA (trimethylsilyl-Ala), alie (alo-isoleucine), NIe (norleucine), fert- Leu, Cit (citrulline), Orn, Dpm (2,2'-diaminopimelic acid), Dpr (2,3-diaminopropionic acid), α- or β-Nal, Cha (cyclohexyl-Ala), hydroxyproline, Sar ( sarcosine), and the like; cyclic amino acids; amino acids N -alkyl such as MeGly (Na-methylglycine), EtGly (Na-ethylglycine) and EtAsn (N ^ -ethylaparagine); and amino acids in which carbon a has two side chain substituents, "Amino acid equivalent" means an amino acid which can be substituted by another amino acid in the peptides according to the invention, without appreciable loss of function. In making such changes, similar amino acid substitutions are made on the basis of relative similarity of side chain substituents, for example in size, charge, hydrophilicity, hydropasticity and hydrophobicity as described herein. As detailed in the patent of E.U.A. No. 4,554,101, incorporated herein by reference, the following hydrophilicity values have been assigned to amino acid residues: Arg (+3.0); Lys (+3.0); Asp (+3.0); Glu (+3.0); Ser (+0.3); Asn (+0.2); Gln (+0.2); Gly (0); Pro (-0.5); Thr (-0.4); Ala (-0.5); His (-0.5); Cys (-1.0); Met (-1.3); Val (-1.5); Leu (-1.8); lie (-1.8); Tyr (-2.3); Phe (-2.5); and Trp (-3.4). It is understood that an amino acid residue can be substituted by another having a similar hydrophilicity value (eg, within a value of plus or minus 2.0) and still obtain a biologically equivalent polypeptide. Similarly, substitutions can be made on the basis of similarity in the hydropathic index. A hydropathic index has been assigned to each amino acid residue based on its hydrophobicity and loading characteristics. Those values of hydrophilic index are: lie (+4.5); Val (+4.2); Leu (+3.8); Phe (+2.8); Cys (+2.5); Met (+1.9); Wing (+1.8); Gly (-0.4); Thr (-0.7); Be (-0.8); Trp (-0.9); Tyr (-1.3); Pro (-1.6); His (-3.2); Glu (-3.5); Gln (-3.5); Asp (- 3.5); Asn (-3.5); Lys (-3.9); and Arg (-4.5). When making a substitution based on the hydropathic index, a value is preferred within plus or minus 2.0. "Peptide" and "polypeptide" mean a polymer in which the monomers are amino acid residues bound through amide bonds. Preferred peptides of the present invention are those comprising amino acids a. The most preferred peptides of the present invention comprise amino acids a having stereochemistry L on carbon a. ™ "Cyclic peptide" means a peptide that contains one or more cyclic peptide fragments as defined herein. "Peptide fragment" means a peptide subunit of the origin peptide. "Peptide fragment" may involve a linear, branched or cyclic subunit of the target peptide. "Linear peptide" means a peptide or polypeptide in which the amino acids are linked to one another through an amide bond between the 9 N-terminal one and the C-terminal another. "Branched peptide" means a peptide or polypeptide within which one or more individual amino acids constituting carboxylic acid or amine side chains are attached to another peptide substituent via these side chains. "Cyclic peptide fragment" means a peptide fragment as defined herein in which a substituent on the residue of amino acid is attached to a substituent on another amino acid residue in the peptide fragment. The preferred link is between the side chains of two amino acid residues in the peptide fragment, preferably through an ester, amide, disulfide or lanthionine linkage. The union between the two amino acid side chains is designated in present as I I. The ester, amide, disulfide or lanthionine linkage, which binds two amino acid residues of the cyclic peptide, is formed between the side chain functionalities. Thus, an amide bond is formed between the side chain carboxyl group of an acidic amino acid residue and the side chain amino group of a basic amino acid residue; the ester linkages are formed between the side chain carboxyl group of an acidic amino acid residue and the side chain hydroxyl group of a hydroxyl-containing amino acid residue; disulfides are formed from amino acid residues containing side chain sulfhydryl groups; and the lanthionine bridges are formed by desulfurization of the corresponding disulfide. The number of atoms in the bridge resulting from the amide, ester, disulfide or lanthionine linkage formed as described above, will vary depending on the length of the side chain and the type of bond (i.e., amide, ester, disulfide or lanthionine) ). Preferably, the bridge comprises from 2 to 12 atoms, preferably from 6 to 10 atoms. The number of atoms of higher preference contained in the bridge is 7, this bridge preferably comprises an amide linkage between the side chain functionalities of a Lys residue and an Asp residue. "Resin" means a solid support modified with a reactive group so that the solid support can be coupled to the carboxy or N-terminus of an amino acid, peptide or cyclic peptide fragment as defined herein. Representative resins include Merrifield resins (chloromethylated polystyrene), hydroxymethyl resin, 2-chlorotrityl chloride resin, trityl chloride resin, Rink acid resin (4-benzyloxy-2 'resin, 4'-dimethoxybenzhydrol) , trityl alcohol resin, PAM resin (4-hydroxymethyl-phenylacetamidomethyl resin), Wang resin (p-benzyloxybenzyl alcohol resin), MBHA resin (p-methylbenzhydrylamine resin), BHA resin (benzhydrylamine resin), Rink amide resin (4- (2 ', 4'-dimethoxyphenyl-Fmoc-amnomethyl) phenoxy resin) and PAL resin (5- (4-) resin Fmoc-aminomethyl-3,5-dimethoxyphenoxy) valeric-MBHA). Preferred resins are chlorotryril resin, Rink acid resin and Rink amide resin. "Solid support" means a substrate which is inert to the reactants and reaction conditions described herein, likewise, it is substantially insoluble in the media used. Representative solid supports include inorganic substrates such as kieselguhr, silica gel and controlled porous glass; organic polymers including polystyrene, which includes 1-2% copolystyrene-divinylbenzene (in the form of gel) and 20-40% of copolystyrene-divinylbenzene (in macroporous form), polypropylene, polyethylene glycol, polyacrylamide, cellulose and the like; and composite inorganic / polymeric compositions such as polyacrylamide supported within a matrix of kieselguhr particles. See J.M. Stewart and J.D. Young, Solid Phase Peptide Synthesis, 2a. Ed., Pierce Chemical Co. (Chicago, IL, 1984). In addition, "solid support" includes a solid support as described above which is attached to a second inert support such as the pins described in Technical Manual, Multipin ™ SPOC, Chiron Technologies (1995) and references thereof, which comprise a head made of polyethylene or removable polypropylene grafted with an amino functionalized methacrylate copolymer and an inert rod. In addition, "solid support" includes polymeric supports such as the polyethylene glycol supports described by Janda et al., Proc. Nati Acad. Sci. USA, 92, 6419-6423 (1995) and S. Brenner, WO 95/16918, which are soluble in many solvents but can be precipitated by the addition of a precipitating solvent. The names of natural and non-natural amino acids and residues thereof used herein, follow the naming conventions suggested by the IUPAC Commission on the Organic Chemistry Nomenclature and the IUPAC-IUB Commission on Biochemical Nomenclature as set forth in "Nomenclature of a-Amino Acids (Recommendations, 1974) "Biochemistry, 14 (2), (1975). To the point where the names and abbreviations of amino acids and residues thereof used in this specification and appended claims differ from those mentioned, will clarify the names and abbreviations that differ. F A representative peptide prepared according to The process of this invention is expressed, for example, as the cycle (Lys18-Asp22) [Ala1, NIe8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 with the amino acid residues joined in the parentheses after "cycle" "and with substituted amino acids of the natural sequence placed in square brackets. hPTH means human PTH and PTHrP means protein related to human parathyroid hormone. The numbers in the second parentheses refer to the number of amino acid residues in the peptide, starting at the N-terminus (ie, the first 31 amino acids of hPTH).

PREFERRED MODALITIES 15 As shown in scheme 1, the preparation of a cyclic f-peptide linked to resin involves the coupling of cyclic and non-cyclic peptide fragments to a resin, amino acid bound to resin or peptide bound to resin. 20 SCHEME 1 ^ 0 PHN-OC02H -E '^ 0 'is a suitable resin for peptide synthesis. T is an H2N, and amino acid, or a cyclic or acyclic peptide. E and O are each, or independently, a protected N-terminal amino acid or a protected N-terminal acyclic or cyclic peptide fragment. The coupling is preferably carried out between the carboxy terminus of the peptide fragment and the resin, amino acid bound to resin or peptide bound to resin. When the peptide fragment is coupled to an amino acid linked to resin or peptide, preferably the coupling is through an amide bond between the carboxy terminus of the cyclic peptide fragment and the N-terminus of the resin-bound amino acid or peptide. In order for the coupling reaction to proceed, the carboxyl group of the peptide fragment must be activated. Many activation methods can be used in the practice of the invention which include, for example, preformed symmetric anhydrides (PSA), preformed mixed anhydride (PMA), acid chlorides, active esters and in situ activation of the carboxylic acid, as established in Fields and Noble, 1990, "Solid phase peptide synthesis utilizing 9-fluorenylmethoxycarbonyl amino acids", Int. J. Pept. Protein Res. 35: 161-214. Representative activating agents include isopropyl chloroformate, diisopropylcarbodiimide (DIC), DIC mixed with 1-hydroxybenzotriazole (HOBT), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide (EDC), bis (2-oxo-3-oxazolidinyl) phosphonic chloride (BOP-CI), benzotriazol-1-yloxy-tris ((d -methylamino) phosphonium hexafluorophosphate (BOP), benzotriazol-1-yloxy-tris-pyrrolidino hexafluorophosphate -phosphonium (PyBOP), bromo-tris-pyrrolidino-phosphonium hexafluorophosphate (PyBROP), 0- (7-azabenzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium hexafluorophosphate (HATU), tetrafluoroborate 2 (1 H-benzotriazol-1-yl) -1.1.3.3-tetramethyluronium (TBTU), 2- (1 H-benzotriazol-1-yl) -1.1.3.3-tetramethyluronium hexafluorophosphate (HBTU), 2- [2] tetrafluoroborate -oxo-1 - (2H) -pyridyl] -1, 2,3,3-bispentamethyleneuronium (TOPPipU), N, N'-dicyclohexylcarbodiimide (DCC), DCC mixed with HOBT and the like Suitable solvents for the coupling reaction they include dichloromethane, DMF, DMSO, toluene, THF, and the like Coupling times range from 1 to about 48 hours, depending on the carboxylic acid derivative and resin to be coupled, activating agent, solvent and temperature. it is carried out at about -10 ° C to about 50 ° C, preferably at about room temperature. In order to avoid interference with the coupling reaction described above, the N-terminus of the cyclic peptide moiety is protected by an acid or sensitive base group. Such protecting groups must have the properties of being stable to the conditions of amide bond formation, and at the same time be easily removable without destruction of the growing chain of peptide or racemization of any of the chiral centers contained therein. The suitable protective groups are 9- fluorenylmethyloxycarbonyl (Fmoc), f-butyloxycarbonyl (Boc), benzyloxycarbonyl (Cbz), biphenylysopropyloxycarbonyl, f-amyloxycarbonyl, isobornyloxycarbonyl, (a.a) dimethyl-3,5-dimethoxybenzyloxycarbonyl, o-nitrophenylsulfenyl, 2-cyano-f-f-butyloxycarbonyl and the like. The protective group 9-5 fluorenylmethyloxycarbonyl (Fmoc) is preferred. Likewise, it may be necessary to protect any reactive side chain functional group in the cyclic peptide fragment. Particularly preferred side chain protecting groups are, for side chain amino groups such as lysine and arginine: 2,2,5,7,8-f-pentamethylchroman-6-sulfonyl (pmc), nitrogen, p-toluenesulfonyl, - methoxybenzenesulfonyl, Cbz, Boc, Alloc (allyloxycarbonyl) and adamantyloxycarbonyl; for tyrosine: benzyl, o-bromobenzyloxycarbonyl, 2,6-dichlorobenzyl, isopropyl, f-butyl (t-Bu), cyclohexyl, cyclopentyl and acetyl (Ac); for serine: f-butyl, benzyl and tetrahydropyranyl; for histidine: trityl, benzyl, Cbz, p-toluenesulfonyl and 2,4-dinitrophenyl; for tryptophan: formyl and Boc; for asparagine and glutamine: Trt (trityl); for aspartic acid and glutamic acid: 0-t-f Bu and OAlilo and Obencilo. Preferably, the coupling of the fragments to the resin-bound peptide or resin is carried out at about room temperature using about 2 molar equivalents (relative to the resin) of the Fmoc-protected cyclic peptide fragment, activated with about a portion equimolar benzotriazole hexafluoroacetate iloxytris [pyrrolidino] -phosphonium (PyBOP) and hydroxybenzotriazole hydrated in the presence of about 4 equivalents of diisopropylethylamine in dimethylformamide for about 16-48 hours. The entire coupling must be evaluated. The experts in ) technique would be familiar with the known monitoring tests such as ninhydrin (the Kaiser test) picric acid, 2,4,6-trinitrobenzenesulfonic acid (TNBS), fluorescamine, and chloranil, which are based on reactive reaction with free amino groups to produce a chromophoric compound. If minoacids are used (eg, Pro and Hyp), isatin monitoring is a preferred method. Fields and Noble, supra. The quantification of the entire reaction can be monitored during the course of the reaction, for example, as described in Salisbury et al. (Patent Publication Interantional No. WO91 / 03485). With the synthesis of Fmoc, the Kaiser and TNBS tests are preferred. In the Kaiser test, a sample of resin peptide with ninhydrin reagent obtained from Pierce Chemical can be tested in the method set forth by Sarin et al. (1981, Anal. Biochem. 117: 147-157). Similarly, a sample of resin peptide can be tested using trinitrobenzenesulfonic acid (TNBS), available from Fluka, as set forth in Takajashi (1984, Chem Lett.1: 127). If the coupling reaction is incomplete, as determined by one or the other of these tests, the reaction may be forced to terminate by several methods known to those skilled in the art, including (a) a repeated coupling using a surplus of one to five times of protected amino acids, (b) a repeated coupling using different solvents or additional solvents (eg, trifluoroethane), or (c) the addition of chaotropic salts, for example, NaCl04 or LiBr (Klis and Stewart, 1990, "Peptides"). : Chemistry, Structure and Biology, "Rivier and Marshall, eds ESCOM Publ., P.904-906). A preferred fragment of cyclic peptide attached to resin has formula II P-A-A18- A19- A20-A21-A22- B-C-D where it is a ream; P is an amine protecting group; A is absent or is a formula group; - A13- A14- A15-A1ff-A17 B is a group of formula - A 23 - A 24 - A 25; C is a group of the formula - A 26 - A 27 - A 2 ß - A 29 - A 30; Y D is A 31 A 31 - A 32 A 31 - A 32- A 33 o A 31 - A 32 - A 33 - A 34 - where it is a bridge of lactam, ester, disulfide or lanthionine; A13, A17, Ais and A22 are amino acid residues; A is His (Trt) or Ser (tBu); A15 and f A28 are independently lie or Leu; Being (tBu) or Leu; A? 6 is Asn (Trt) or Gln (Trt); A19 is Arg (Pmc) or Glu (OtBu); A20 is Arg (Pmc); A21 is Arg (Pmc) or Val; A23 is Phe or Trp (Boc); A2 is Leu; A2s is Arg (Pmc) or His (Trt); A26 is A30 are amino acid residues wherein the side chains of A2β and A3o are optionally linked through an amide, disulfide or lanthionine linkage; A27 is Leu or Lys (Boc); A2g is Ala or Gln (Trt); A31 is lie or Val; A32 is f 10 His (Trt); A33 is Asn (Trt) or Thr (tBu); and A ^ is Ala or Phe. Another preferred fragment of cyclic peptide attached to resin has formula II wherein 1 'is a lactam bridge and the side chains of A26 and A30 are optionally linked through an amide bond. Another cyclic peptide linked to preferred resin has formula II wherein it is a lactam bridge; A13 and A? 8 are Lys; A17 and A22 are 20 Asp; and A26 is Lys (Boc); and A 0 is Asp (OtBu); or the side chains of A26 and A30 are optionally linked through an amide bond. Another preferred fragment of cyclic peptide attached to resin has formula III Fmoc- A18- AIG- A20- A21- A22- A23- A24- A25- A26- A27- A28- A29- A3Q- A3 where 1 'is a lactam bridge; A18 is Lys; A19 is Glu (OtBu); A or is Arg (Pmc); A21 is Val; A22 is Asp; A23 is Trp (Boc); A2 is Leu; A25 is Arg (Pmc); A2ß is Lys (Boc); A27 is Leu; A28 is Leu; A2g is Gln (Trt); A30 is The resin-bound peptide prepared as described above can be further elaborated, for example by coupling with one or more additional cyclic peptide fragments, by coupling with one or more peptide fragments, by consecutive addition of individual amino acids or by any combination of the above. Then, the complete cyclic peptide is cleaved from the resin and purified. The protecting groups can be removed before or after or simultaneously with the resin segmentation. The completely deprotected peptide can be purified, alone or in combination, by acid-base extraction, recrystallization, lyophilization, by a sequence of chromatographic steps employing any or all of the following types: ion exchange in a weakly basic resin in the form of acetate; hydrophobic absorption chromatography on non-derived polystyrene-divinylbenzene (e.g., AMBERLITE® XAD); chromatography of silica gel adsorption; and ion exchange chromatography on carboxymethylcellulose; division chromatography, for example in SEPHADEX® G-25, LH-20 or countercurrent distribution; high performance liquid chromatography (HPLC), especially reversed phase HPLC in pack phase column attached to octyl-or octadecylsilyl-silica.

Preparation of the peptide fragment The fragment is prepared by synthesizing the base structure of the peptide, which causes the assembly of the amino acids comprising the fragment in its proper order. The peptide base structure of the fragment can be synthesized by any of the techniques that are known to those skilled in the art. For synthesis of solid phase peptide, a summary of the different techniques can be found in J.M. Steward and J.D. Young, Solid Phase Peptide Synthesis, W.H. Freeman Co. (San Francisco), 1963 and J. 15 Meienhofer, Hormonal Proteins and Peptides, vol. 2 P. 46, Academic Press (New York), 1973. For classical solution synthesis see G. Schroder and K. Lupke, The Peptides, vol. 1, Academic Press (New York), 1965. As is known to those skilled in the art, the method of peptide synthesis in solid supports generally involves the construction of a peptide from the carboxyl or C-terminal end in the which the C-terminal amino acid with its protected a-amino group binds to a solid phase polymer. The N-protecting group is then cleaved and the next amino acid, also N-protected, is coupled via a link to peptide to the a-amino group of the amino acid attached to the solid support as described above. The deprotection cycle of the above amino acid and the coupling of the additional amino acid is repeated until the peptide is complete. Any reactive side chain of the amino acids, is protected by chemical groups that can support the coupling procedure and Na-deprotection but that can be removed at the end of the synthesis. When the peptide fragment is prepared using solid phase methods, the fragment is obtained for use in couplings to other fragments by cleavage of the resin. A cyclic fragment is prepared in a similar manner. However, the assembly of the base structure is followed by (1) selective deprotection of the side chain functionalities that will be cyclized, (2) cyclization and (3) optional removal of any remaining protecting group. The use of Fmoc amino acids is only one strategy of peptide synthesis. A Boc (t-butyloxycarbonyl-protected amino group) strategy can also be used to prepare a peptide bond to the solid support (eg, Geysen et al., 1987, J. Immunol. 102: 259-274). The amino acids used for peptide synthesis can be standard Boc amino acids (protected Na-t-butyloxycarbonyl Na-amino) described by Merrifield (1963, J. Am. Chem. Soc. 85: 2149-2154), or the amino acids of 9- fluorenylmethoxycarbonyl Na-amino protected labile bases first described by Carpino and Han (1972, J. Org. Chem. 37: 3403-3409). The protected Fmoc and Boc Na-amino amino acids can be obtained from Fluka, Bachem, Advanced Chemtech, Sigma, Cambridge Research Biochemical, Bachem, or Peninsula Labs or other companies? Chemicals related to those that practice this technique. In addition, the The method of the invention can be used with other Na-protecting groups that are known to those skilled in the art. Solid phase peptide synthesis can be performed by techniques known to those skilled in the art and provided, for example, in Stewart and Young, 1984, Solid Phase Synthesis, Second Edition, Pierce Chemical Co., Rockford, IL; Fields and Noble, 1990, Int.

H? J. Pept. Protein Res. 35: 161-214, or using automated synthesizers, such as those sold by ABS. Although the synthesis of C-terminal peptide to N-terminal is normal, those skilled in the art will recognize that the peptide fragment can also be synthesized from N-terminal to C-terminal. Preparation of the lactam bridge The lactam-linked cyclic peptide fragments are prepared by the formation of an amide bond between the side chain carboxyl group of an acidic amino acid residue and the side chain amino group of an amino acid residue. basic in the presence of an activating agent as described above. Preferred acidic amino acid residues include Asp, Glu, -NHCH [(CH2) 3C02H] CO- and -NHCH [(CH2) 4C? 2H] CO-, with Asp being most preferred. The waste Preferred basic amino acids include His, Lys, Orn, NHCH (CH2NH2) CO- and -NHCH [(CH2) 2NH2] CO-, Lys being most preferred. In cases where the peptide precursor to the cyclic peptide fragment contains more than one acidic or basic amino acid residue, the protecting groups for additional basic or acidic amino acids in the cyclic peptide fragment are selected so that the amino acids which will be cyclized can be selectively deprotected. Preferably, the desired acidic and basic amino acid residues are simultaneously deprotected. Furthermore, in addition to being stable to the reagents used to deprotect selected residues of basic and acidic amino acids, the protecting groups on the remaining amino acid residues are selected to be stable under the cyclization conditions employed. The term "orthogonality" when used in reference to side chain protecting groups, refers to a situation as described herein, where there are two or more kinds of protecting groups in a molecule, each class optimally removed under specific conditions , although remaining stable to conditions used to remove protective groups in other classes. In this way, all the protective groups of one class can be removed, while all the others remain intact.

The preferred protecting groups having the desired orthogonality are: for the acidic amino acid residue that will be cyclized: allyl; for the basic amino acid residue that will be cyclized: allyloxycarbonyl f (alloc); for any additional acidic amino acid residue: terbutyl (tBu); and for any additional basic amino acid residue: feroutyloxycarbonyl (Boc) After synthesis of the peptide base structure of the cyclic peptide fragment using solution phase or solid phase techniques, the allyl and allyloxycarbonyl protecting groups are simultaneously removed by treatment with palladium, preferably tetrakis (triphenyl). Lphosphine) palladium (O) in the presence of N-methylaniline. The formation of the lactam bridge is then performed as described herein for amide bond formation. Preferred cyclic peptide fragments for use in the preparation of the resin bound cyclic peptides of this invention have formula XV, XVI or XVII P- A? 3- A14- AI5-A16-A17- OH XV 20 P- Ai 8- Ai 9- A20-A21-A22- OH XVI P- A26- A27- A28-A29- A ^ - OH XVII where I I is a lactam, disulfide or lanthionine bridge; P is an amine protecting group; A? 3, A17, A? ß, A22, A26 and A30 are amino acid residues; Au is His (Trt) or Ser (tBu); A15 is Ser (tBu) or Leu; A? 6 is Asn (Trt) or Gln (Trt); A19 is Arg (Pmc) or Glu (OtBu); A20 is Arg (Pmc); A2? is Arg (Pmc) or Val; A2 is Leu or Lys (Boc); A28 is lie or Leu; and A2g is Ala or Gln (Trt). The most preferred cyclic peptide fragments have formula IV, V or VI where '' is a lactam bridge; P is Fmoc; A? 3, Ais and A2? Are Lys; A? 7, A22 and A30 are Asp; Au is His (Trt) or Ser (tBu); A15 is Ser (tBu) or Leu; A? 6 is Asn (Trt) or Gln (Trt); A19 is Arg (Pmc) or Glu (OtBu); A2o is Arg (Pmc); A21 is Arg (Pmc) or Val; A27 is Leu or Lys (Boc); A2s is ile or Leu; and A2g is Ala or Gln (Trt). A cyclic peptide fragment even more preferred for use in the preparation of the resin bound cyclic peptides of this invention is Fmoc-cyclo (Lys-Asp) Lys-Glu (OtBu) -Arg (Pmc) -Val-Asp-OH .

Cyclic peptide parathyroid hormone analogs and parathyroid hormone related protein analogs suitable for the preparation using the method of this invention are described in: 1. Condón et al., Series of E.U.A. No. 60 / 046,472 (filed May 14, 1997); 2. Willick et al., Patent of E.U.A. No. 5,556,940 (September 17, 1997); 3. Chorev et al., Patent of E.U.A. No. 5,717,062 (February 10, 1998); 4. Vickery et al., WO 96/40775 (published December 19, nineteen ninety six); A preferred cyclic peptide suitable for the preparation using the process of this invention has formula I X-A7-A8-Ag-A1o-A? 1-A? 2-Ai3-Ai4-A15-A16-Ai7-A18-A? g-A2o-A21- A22-A23-A24-A25-A26-A27-A28-A2g-A3o-A3i-YI wherein X is selected from the group consisting of (a) R? aA? -A2-A3-A4 -A5-A6- (b) Rib-A2-A3-A4-A5-A6-, (c) R ^ -As-rAs-Ae-, (d) R1b-A4-A5-A6-, (e) R β-A5-A6-, (f) Rib-Aβ- and (g) Rib- wherein Rαa is selected from the group consisting of (1) H, (2) a peptide comprising from 1 to 6 amino acids , (3) alkyl, (4) phenylalkyl, (5) -COR2 wherein R2 is selected from alkyl, alkenyl, alkynyl, phenyl, naphthyl and phenylalkyl, Rib is selected from the group consisting of (1) H, (2) alkyl, (3) phenylalkyl and (4) -COR2 wherein R2 is selected from alkyl, alkenyl, alkynyl, phenyl, naphthyl and phenylalkyl, Ai is selected from Ser and Ala or a residue of amino acid equivalent thereof, A2, A3, A4, and Ae, are amino acid residues and A5 is selected from lie and His or an equivalent amino acid residue thereof; A7, Ag, An and A12 are amino acid residues; As is Leu or NIe or an equivalent amino acid residue thereof; A10 is Asp or Asn or an equivalent amino acid residue thereof; A13 and A17 are amino acid residues wherein the side chains of A13 and A17 are linked through an amide bond. Au is His or Ser or an equivalent amino acid residue thereof; A15 and A28 are independently either Lie or Leu or an equivalent amino acid residue thereof. A16 is Asn or Gln or an equivalent amino acid residue thereof; Ais and A22 are amino acid residues wherein the side chains of A? 8 and A2 are linked by an amide bond; A19 is Arg or Glu or an equivalent amino acid residue thereof; A20 is Arg or an equivalent amino acid residue thereof; A2? is Arg or Val or an equivalent amino acid residue thereof; A23 is Phe or Trp or an amino acid residue equivalent to the same; A24 is Leu or an equivalent amino acid residue thereof; A25 is Arg or His or an equivalent amino acid residue thereof; A2e and A3o are amino acid residues wherein the side chains of A26 and A3o are optionally linked through a f-amide bond; A27 is Leu or Lys or an equivalent amino acid residue thereof; A29 is Ala or Gln or an equivalent amino acid residue thereof; 15 A3? is lie or Val or an equivalent amino acid residue thereof; f Y is selected from the group consisting of (a) R3, (b) A32-R3, (c) - A32A33, -R3 and (d) -A32-A33-A; 34-R3 wherein R3 is -OH or -NR4R5 wherein R and R5 are independently selected from hydrogen and lower alkyl, A32 is His or an equivalent amino acid residue thereof, A33 is Asn or Thr or an equivalent amino acid residue thereof and A34 is Ala or Phe or an equivalent amino acid residue thereof.

The peptides of formula I possess useful pharmaceutical properties. They are especially useful for treating disease states capable of being modulated by compounds that bind to parathyroid hormone receptors with or without concomitant stimulation of cAMPase activity. See E.U.A. Serial No. 60 / 046,472. Representative preferred cyclic peptides for preparation using the method of this invention include, but are not limited to Ala1, Nle8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 Ala1'2, Nle8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 Ala1'3, Nle8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 Ala1 ', Nle8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 Ala1'5, Nle8, Lysl8 , Asp22, Leu27] hPTH (1-31) NH2 Ala? e, Nle8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 Ala1'7, Nle8, Lysl8, Asp22, Leu27] hPTH (1-31) NH2 Ala1'8, Lys18, Asp22, Leu27] hPTH (1 -31) NH2 Ala1'10, Nle8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 Ala1'11, Nle8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 Ala1 '2, Nle8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 Ala1'13, Nle8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 Ala1'14, Nle8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 Ala1 '15, Nle8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 Ala1 '6, Nle8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 bicyclo (Lys13-Asp17, Lys18-Asp22) [Ala1, Nle8, Lys18, Asp17'22, Leu27] hPTH (1-31) NH2 bicyclo (Lys18-Asp22, Lys26-Asp30) [Ala1, Nle8, Lys18, Asp22, Leu27 ] hPTH (1-31) NH2 cycle (Lys18-Asp22) [Ala1, Nle8, Lys18, Asp22, Leu27] hPTH (1-34) NH2 bicyclo (Lys13-Asp17, Lys18-Asp22) [Ala1, Nle8, Lys18, Asp17 '22, Leu27] hPTH (1-34) NH2 bicyclo (Lys18-Asp22, Lys26-Asp30) [Ala1, Nle8, Lys18, Asp22, Leu27] hPTH (1-34) NH2 cycle (Lys18-Asp22) [Lys18, Asp22 ] hPTHrP (1-34) NH2 cycle (Lys18-Asp22) [Lys18'26'30, Asp22, Leu23'28'31, Glu25'29] hPTHrP (1-34) NH2 cycle (Lys18-Asp22) [Nle8, Lys18 , Asp22, Leu27] hPTH (7-34) NH2 cycle (Lys18-Asp22) [Lys18, Asp22] hPTHrP (7-34) NH2 bicyclo (Lys13, Asp17, Lys 8-Asp22) [Nle8, Lys18, Asp22, Leu27] hPTH (7-34) NH2, and bicyclo (Lys18-Asp22, Lys2β-Asp30) [Nle8, Lys18, Asp22, Leu27] hPTH (7-34) NH2. Another preferred cyclic peptide suitable for preparation using the method of this invention has formula I wherein the amino acid residues are amino acid residues that have stereochemistry L on carbon a.

Another preferred cyclic peptide suitable for preparation using the process of this invention has formula I wherein X is selected from the group consisting of f (a) Ria-A? -A2-A3-A4-A5-A6- wherein R? A is H or Pro, Ai is Ser or Wing, A2 is Val, A3 is Ser, A4 is Glu, A5 is He or His, Aβ is Gln, (b) R1b -A2-A3- A4-A5-A6- where A2, A3, A », A5 and A6- are defined above and R ^ is H, (c) Rib-A3-A4-A5-A6- wherein R? b, A, A4, A5 and Aβ are defined above, (d) Rib-At-As-Aß- where Rib, A4, A and A6 are defined above, (e) R? B-A5- A6- where Rib, A5 and A6 are defined above, (f) Ri Aß- in where Rib yf 10 A6 are defined above and (g) R ^ - wherein R? b is defined above; A7 is Leu; A8 is Leu or NIe; A9 is His; A10 is Asp or Asn; An is Leu; A12 is Gly; A13, A17, A? 8, A22 and A30 are independently selected from Ser, Thr, Lys, Cys, homo-Cys Orn, Asp, Glu, -NHCH (CH2NH2) CO-, - NHCH [(CH2) 2NH2] CO-, -NHCH [(CH2) 3C02H] CO- and -NHCH [(CH2) 4C02H ]CO-; Au is His or Ser; A15 is Ser or Leu; A? 6 is Asn or Gln; A19 is Arg or Glu; A2o is Arg; A21 is Arg or Val; A23 is Phe or Trp; A24 is Leu; A25 is Arg or His; A26 is f His or is independently selected from the group consisting of Ser, Thr, Lys, Cys, homo-Cys Orn, Asp, Glu, -NHCH (CH2NH2) CO-, - NHCH [(CH2) 2NH2] CO-, -NHCH [(CH2) 3C02H] CO- and -NHCH [(CH2) 4C02H ]CO-; A27 is Leu or Lys; A28 is lie or Leu; A2g is Ala or Gln; A31 is lie or Val; and Y is selected from the group consisting of (a) -R3 wherein R3 is -OH or NR4R5 wherein R4 and R are independently selected from hydrogen and alkyl of one to four carbon atoms, (b) -A32-R3 in where R3 is defined above and A32 is His, (c) -A32-A33-R3 wherein R3 and A32 are defined above and A33 is Asn or Thr and (d) -A32-A33-A34 wherein A ^ is Ala or Phe. f Another preferred cyclic peptide suitable for preparation using the process of this invention has formula I wherein (i) the side chains of Aβ8 and A22 are linked by an amide bond, (ii) the side chains of A13 and A17 are linked by an amide bond, and f the side chains of Aβ8 and A22 are linked by an amide linkage, (iii) the side chains of Ais and A22 are linked by an amide bond, and the side chains of A26 and A3o are linked by an amide bond. amide bond or (iv) the side chains of A13 and A17 are linked by f an amide bond, the side chains of Aia and A22 are linked by an amide bond, and the side chains of A26 and A30 are linked by an amide bond. Another preferred cyclic peptide suitable for preparation using the method of this invention has formula I wherein A? 3 is select from Lys and Ala, A17 is selected from Ser and Asp, Ais is Lys, A22 is Asp, A26 is Lys, and A3o is Asp. Another preferred cyclic peptide suitable for preparation f using the process of this invention has formula I wherein X is Ria-A? -A2-A3-A4-A5-As-. Another preferred cyclic peptide suitable for preparation using the method of this invention has formula I wherein Ai is Ala, A8 is NIe and A27 is Leu. Another preferred cyclic peptide suitable for preparation using the process of this invention has formula I wherein R a is H and Y is NH 2. Another preferred cyclic peptide suitable for preparation using the process of this invention has formula I wherein R? A is H and Y is-A32-A33-A34-NH2. Another preferred cyclic peptide suitable for preparation using the method of this invention has formula I wherein X is selected from the group consisting of (a) Rib-A2-A3-A4-A5-A6-, (b) Rib-A3- A4-A5-A6-, (c) Rib- t-As-Ae-, (d) Rib-A5-A6-, (e) RuAr, and (f) R? -. Another preferred cyclic peptide suitable for preparation using the method of this invention has formula I wherein Aβ is NIe and A27 is Leu.

Another preferred cyclic peptide suitable for preparation using the method of this invention has formula I wherein X is H and Y is - f More preferred cyclic peptides suitable for preparation using the method of this invention are selected from cycle (Lys18-Asp22) [Ala1, NIe8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 cycle (Lys 8-Asp22) [Ala1'2, NIe8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 cycle (Lys18-Asp22) [Ala1'3, NIe8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 cycle (Lys18-Asp22) [Ala1'4, NIe8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 i 10 cycle (Lys18-Asp22) [Ala1'5, NIe8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 cycle (Lys18-Asp22) [Ala1 ' 6, NIe8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 cyclo (Lys18-Asp22) [Ala1'7, NIe8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 cycle (Lys18-Asp22) [ Ala1'8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 cycle (Lys18-Asp22) [Ala1 10, NIe8, Lys18, Asp22, Leu27] hPTH (1- 15 31) NH2 cycle (Lys18-Asp22) [ Ala1'11, NIe8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 cycle (Lys18-Asp22) [Ala1'12, NIe8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 cycle (Lys18- Asp22) [Ala1'13, NIe8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 cycle (Lys18-Asp 2) [Ala1'14, NIe8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 cycle (Lys18-Asp22) [Ala1'15, NIe8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 cycle (Lys18-Asp22) [Ala1'16, NIe8, Lys18, Asp22, Leu27] hPTH (1-31 ) NH2 bicyclo (Lys13-Asp17, Lys 8-Asp22) [Ala1, Nle8, Lys18, Asp17'22, Leu27] hPTH (1-31) NH2 bicyclo (Lys18-Asp22, Lys28-Asp30) [Ala1, Nle8, Lys18, Asp 2, Leu27] hPTH (1-31) NH2 Even more preferred cyclic peptides suitable for preparation using the method of this invention are selected from cycle (Lys18-Asp22) [Ala1, Nle8, Lys18, Asp22, Leu27] hPTH (1- 31) NH2 bicyclo (Lys13-Asp17, Lys18-Asp22) [Ala1, Nle8, Lys18, Asp17'22, Leu27] hPTH (1-31) NH2 bicyclo (Lys18-Asp22, Lys26-Asp30) [Ala1, Nle8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 An even more preferred cyclic peptide suitable for preparation using the method of this invention is cyclo (Lys18-Asp22) [Ala1, Nle8, Lys18, Asp22, Leu27] hPTH (1-31) NH2. The foregoing can be better understood by reference to the following example, which is presented for illustration and is not intended to limit the scope of this invention.

EXAMPLE 1 Preparation of cycle (Lvs18-Asp22) rAla1.NIe8, Lvs1B.Asp22.Leu271hPTH (1- 3DNH, Ala-Val-Ser-Glu-lle-Gln-Leu-Nle-His-Asn-Leu-Gly-Lys-His-Leu-As? Ser-Lys-Glu-Arg-Val-Asp-Trp-Leu-Arg-Lys-Leu-Leu-GIrvAsp-Val-NH 2 Step 1: Resin of Fmoc-Trp (Boc) -Leu-Arg (Pmc) -Lys (Boc) -Leu-Leu-f 10 Gln (Trt) -Asp (OtBu) -Val-NH. Rink Amida MBHA resin (0.5-0.7meq / g titration, Nova Biochem, LaJolla, CA, USA), 5 g (0.64 mmoles / g, 3.2 mmoles) is swollen by shaking with 60 mL of dimethylformamide, then the resin is drained and washed with two 60 mL portions of dimethylformamide. The group 9-Fluorenylmethoxycarbonyl (FMOC) is then removed by treating the resin with 60 mL of 20% piperidine / dimethylformamide for 20 minutes, followed by f 4 washes of 60 mL of the resin with dimethylformamide (TNBS: positive) Fmoc-Val-OH (2.17 g, 6.4 mmoles) and 2.43 g (6.4 mmoles) of O-benzotriazol-1-yl-N, N, N ', N'-tetramethyluronium hexafluorophosphate (HBTU) in 150 mL of dimethylformamide are treated with 32 mL (12.8 mmol) of N-methylmorpholine a 0 ° C for 10 minutes, then the mixture is added to the resin peptide and the mixture is stirred at room temperature for about 1.3 hours. The resin was drained, then washed with three 150 mL portions of dimethylformadia (TNBS: negative). The resin was washed with two portions of diethyl ether and dried overnight; the weight gain of the coupling was 0.88 g, implying the addition of 2.75 mmoles of Val. The synthesis was followed by consecutive removal of FMOC (as above) and coupling (as above) with 5.6 mmol each of FMOC-Asp (OtBu), FMOC-Gln (Trt), FMOC-Leu, FMOC-Leu, FMOC-Lys (BOC), FMOC-Arg (PMC ), FMOC-Leu, and FMOC-Trp (BOC), using 5.6 molar equivalents of HBTU, 11.2 mmoles of N-methylmorpholine and 50 mL of dimethylformamide in each coupling. The final weight gain was 5.8 g. f 10 An aliquot was segmented to provide the peptide of the titer that retains the FMOC but with unprotected side chains, 86A% purity. LC-MS: 1390.6; cale 1390.8).

Step 2: 15 Fmoc-cycle (Lys-Asp) Lys-Glu (OtBu) -Arg (Pmc) -Val-Asp-OH.

Fmoc-Lys- Qu (OtBu) - A? P (Pmc) - Val- Asp- OH Method A: Solid phase synthesis. to. Sequence assembly: Resin synthesis of Fmoc-Lvs (Alloc) -Glu (OtBu) -Arq (Pmc) -Val-AspfOAIIyl) -Clorotritil. The coupling of Fmoc-Asp (OAIIyl) -OH with 15 g of chlorotrityl resin crosslinked at 1% loading of 1.05 meq / g (total 15.75 mmol) was carried out using 7.89 g (18.9 mmol) of Fmoc-Asp ( OAIIyl) -OH, 82 g (39.9 mmol) of benzotriazolyloxy-tris [pyrrolidino] -phosphonium hexafluoroacetate (PyBOP) and 10.2 g (78.8 mmol) of diisopropylethylamine in 150 mL of dichloromethane. The mixture was stirred, then drained and the resin was washed using three 150 mL portions of dichloromethane / methanol / diisopropylethylamine. (17: 2: 1), followed by three 150 mL portions of dichloromethane. A sample of the resin was obtained for a TNBS test, which was negative. The N-terminal 9-fluorenylmethoxycarbonyl group was removed by treating the amino acid bound to Fmoc-protected resin with 150 mL of 5% piperidine in dichloromethane-dimethylformamide (1: 1) for 10 minutes followed by 150 mL of 20% piperidine in dimethylformamide. for 30 minutes. The resin bound amino acid was washed as described above. Fmoc-Val-OH, Fmoc-Arg (Pmc) -OH (10% aliquot removed after this step), Fmoc-Glu (OtBu) -OH and Fmoc-Lys (e-Alloc) -OH are each coupled in the same manner and using the Same sequence of events however using three to five washes of dimethylformamide instead of three dichloromethane washes before the TNBS test. No couplings are required double. The peptide bound to final resin was washed with 150 mL of tetrahydrofuran and 150 mL of diethyl ether, then dried under vacuum to yield 27.23 g. b. Removal of Allyl and Alloc for Lvs18 v Asp22: Resin synthesis of Fmoc-Lvs-Glu (OtBu) -Arq (Pmc) -Val-Asp-Chlorotrityl. A 10 g portion of the Fmoc- (Lys-Alloc) -Glu (OtBu) -Arg (Pmc) -Val-Asp (OAIIyl) -chlorotryl resin prepared above was swollen on a shaker with 100 mL of dimethylsulfoxide / dimethylformamide / dichloromethane (1: 1: 0.2 v / v / v) for 20 minutes, then the resin was gently drained. A partial solution of 2.9 g of tetrakistriphenylphosphinpalladium (0) in 240 mL dimethylsulfoxide / dimethylformamide (1: 1, v: v) was added, followed by 60 mL of dichloromethane and 16.2 g (150 mmol) of N-methylanaline, then mixing it was stirred for 2 hours. The resin was drained and washed with 3 portions of 100 mL of dichloromethane. Then the resin was swollen with 100 mL of dimethylsulfoxide / dimethylformamide (1: 1 v: v), then drained gently and used directly in Lys18-Asp23 cyclization. c. Formation of Lactama Bridge of Lvs18-Asp22: Synthesis of Fmoc-cycle resin (Lvs-Asp) Lvs-Glu (OtBu) -Arq (Pmc) -Val-Asp-Chlorotrile. The resin peptide of the previous example is treated with 5.2 mg (10 mmol) of benzotriazolyloxytris [pyrrolidino] -phosphonium hexafluoroacetate (PyBOP) and 2.6 g (20 mmol) of diisopropylethylamine in 100 mL of dimethylsulfoxide / dimethylformamide (1: 1 v: v), and the mixture is stirred for 4 hours. The resin was drained and washed with 3 portions of 100 mL of dimethylformamide, then an aliquot of drained resin was removed for the TNBS test (negative). The resin was washed with 100 mL of each of Tetrahydrofuran and diethyl ether were then dried under vacuum. d. Resin cleavage and product isolation of Fmoc-cycle (Lvs-Asp) Lvs-Glu (OtBu) -Arq (Pmc -Val-Asp-OH The resin peptide of the f-lactam bridge formation example of Lys18-Asp22 The above was treated in an agitator with 130 mL of acetic acid / dichloromethane / 2,2,2-trifluoroethanol (1: 8: 1) for 20 minutes.

The liquid portion was extracted from the resin and treated with 500 mL of methyl tertiary butyl ether and 100 mL of diethyl ether, and the mixture was centrifuged. The supernatant was removed, and diethyl ether was added to the residue, followed by centrifugation. The supernatant was removed and the residue was dried overnight to yield 2.77 g of a yellow solid product (MS: 1172; f 41% stoichiometric yield, crude). This was purified by flash chromaaphy (85:15 to 60:40 dichloromethane: methanol) to yield 1 g of Fmoc-cyclo (Lys-Asp) Lys-Glu (OtBu) -Arg (Pmc) -Val-Asp-OH (15% yield based on resins) with 85A% purity.

Method B: Synthesis in solution. a) Sequence assembly Synthesis of Boc-Asp (OAIIyl) -OBzl In a 3-liter 3-necked purity flask loaded with 1, 1-dimethylethoxycarbonyl- (L) -aspartic acid benzyl ether (50 g, 154.5 mmol), dichloromethane (96 mL), allyl alcohol (9.43 g, 162.3 mmol) and 4-NN-diemthylaminopyridine (10 mg) at room temperature. The reaction mixture H O is cooled to 8 ° C, and a solution of dicyclohexylcarbodiimide (32 g, 154.5 mmol) in dichloromethane (100 mL) is added for 1.5 hours at 8-11 ° C. As the dicyclohexylcarbodiimide solution is added, the dicyclohexylurea precipitates. The reaction mixture is stirred for 2 hours at 11-12 ° C, at which point the HPLC shows no starting material. The dicyclohexylurea is filtered and the filtrate is concentrated. The oily residue is taken up in methyl-t-butyl ether (250 mL), and the solution is filtered to remove residual dicyclohexylurea, washed with 30 mL 0.2 N hydrochloric acid, 30 mL water, 20 mL aqueous sodium bicarbonate saturated, 30 mL of water, and two 50 mL portions of brine, dried (magnesium sulfate), filtered, and concentrated to give 55.88 g of BOC- (L) -Asp (OAIIyl) -OBzl as a oil. (Mcai.363.4; M + 10bsvd364).

Synthesis of HCI-Asp (OAIIyl-OBzl) A solution of BOC-Asp (OAIIyl) -OBzl (110.5 g, 302.7 mmol) in 500 mL of ethyl acetate in a 3-neck 3-L flask with a mechanical stirrer is cooled to 5 ° C and gaseous hydrogen chloride is bubbled through the reaction mixture for 2.5 hours during which the reaction temperature reaches about 22 ° C. CLAR does not show starting material. Nitrogen is bubbled into the solution overnight, which results in the formation of a white precipitate which is collected by filtration and washed with 100 mL of ethyl acetate. The residue is dried under vacuum to yield 84 g of HCl »Asp (OAIIyl) - OBzl as a white crystalline solid product (microscopy). (Mcaic263; M + 10 Svd264).

Synthesis of BOC-Val-Asp (OAIIyl) -OBzl To a solution of BOC-Val-OH (25.4 g, 116.7 mmol) in ethyl acetate (85 mL) is added hydroxybenzotriazole hydrate (16.55 g, 122.5 mmol) in dimethylformamide ( 122.3 mL). The mixture is cooled to 5 ° C and a solution of dicyclohexylcarbodiimide (25.2 g, 122.5 mmol) in ethyl acetate (58.7 mL) is added for 1 hour during which the temperature is raised to about 18 ° C and the dicyclohexylurea is added. precipitates The cooling is terminated and the mixture is stirred for 2 hours and a solution of HCl.H2N-Asp (OAII) -OBzl (35 g, 116.7 mmol) in 189 mL of dimethylformamide is added over 25 minutes. N-methylmorpholine (12.8 L) is add slowly and for 0.5 hours to obtain a reaction mixture pH of 7 and the mixture is stirred at room temperature for 36 hours. The reaction mixture is filtered to remove dicyclohexylurea, the filtrate is diluted with methyl t-butyl ether, stirred for 2 hours, then refiltered. The filtrate is washed with 100 mL of water, 100 mL of 0.5 M aqueous citric acid, 100 mL of water, 100 mL of saturated aqueous sodium bicarbonate, 100 mL of brine, dried (magnesium sulfate), filtered, and concentrated to give 41.3 g of BOC-Val-Asp (OAIIyl) -OBzl as an oil. (Mcaic462; M + 1t, bsVd463).

Synthesis of HCL »Val-Asp (OAIIyl) -OBzl A solution at 2-4 ° C of BOC-Val-Asp (OAN) -OBzl (31.7 g, 68.5 mmol) in ethyl acetate (300 mL) is charged with chloride of gaseous hydrogen for 2 hours. The cooling is finished and the mixture is stirred for 48 hours. Nitrogen is bubbled thoroughly into the solution for 1 hour. The reaction mixture is concentrated on the rotary evaporator to give 38.3 g of a white solid product, which is crystallized from ethyl acetate to yield 24.2 g of HCl »Val-Asp (OAIIyl) -OBzl. (Mcaic362; M + 10bsvd363).

Synthesis of FMOC-ArqfPMC) -Val-Asp (OAIIyl) -OBzl In a 3-L 3-neck flask equipped with a mechanical stirrer are placed 17.25 g (26 mmol) of FMOC-Arg (PMC) and 300 mL of acetonitrile and the mixture is stirred at room temperature until a clear solution is obtained. Benzotriazolyloxytris [pyrrolidino] -phosphonium hexafluoroacetate (PyBOP) is added, the solution is cooled to 2 ° C, and 2.6 f g (26 mmol) of N-methylmorpholine are added, forming a white tar. 5 Dimethylformamide (52 mL) is added slowly until the tar dissolves, and the mixture is stirred at 3 ° C for 30 minutes. A solution of HCI »Val-Asp- (OAIIyl) -OBzl) (10.4 g, 26 mmol) in 39 mL of dimethylformamide is added for 20 minutes at 2-3 ° C, followed by N-methylmorpholine (7.5 mL) to obtain pH 7. The cooling is finished, and the mixture is stirred for M O 2.5 hours. The mixture is then diluted with 250 mL of water and 250 mL of ethyl acetate. The ethyl acetate layer is removed and washed with water (2 x 100 mL) and brine (2 x 100 mL) and concentrated to produce a white solid product which is crystallized from 175 mL of ethyl acetate to give 18.5 g of FMOC-Arg (PMC) -Val-Asp (OAIIyl) -OBzl (95A%). The ethyl acetate filtrate is concentrated and the gummy residue is triturated with ethyl acetate to yield an additional 3.12 g of product. (Mcaic1007; M + 1obsVd1008).

Synthesis of Arq (PMCVVal-Asp (OAIIyl) -OBzl 20 To a solution of FMOC-Arg (PMC) -Val-Asp (OAIIyl) -OBzl (16 g, . 9 mmol in dichloromethane (267 mL) is added piperidine (23 g, 27 mmol, 26.7 mL) at room temperature. The reaction is stirred 2.5 hours and then washed with water (2 x 60 mL) and brine (2 x 60 mL), dried (sulfate magnesium), filtered, and concentrated to give a semi-solid product. The semisolid product is absorbed in 400 mL of ethyl acetate and the solution is left to stand for 3 days and then filtered. The filtrate is washed with water (2 x 60 mL), dried (magnesium sulfate), filtered and concentrated to yield 23.5 g of product contaminated with piperidine-benzofulvene adduct (MS analysis). The solid product is dissolved in 400 mL of ethyl acetate, filtered, washed with water (2 x 60 mL), dried (magnesium sulfate), and concentrated to produce a solid product that is dried overnight under vacuum to yield 12.55 g of Arg (PMC) -Val-Asp (OAII) -OBzl as a white solid product. (Mcaic784; M + 10bsVd785).

Synthesis of FMOC-Glu (OtBu) -Arq (PCM) -Val-Asp (OAIIyl) -Obzl To a solution of FMOC-Glu (OtBu) (6.73 g, 15.8 mmol) and benzotriazolyloxy-tris [pyrrolidino] -phosphonium hexafluoroacetate (PyBOP, 8.18 g, 15.8 mmol) in 105 mL of acetonitrile at 2 ° C is added N-methylmorpholine (1.58 g, 15.8 mmol) and the reaction mixture is stirred at 2-4 ° C for 30 minutes. A solution of L) Arg (PMC) -Val-Asp (OAIIyl) -OBzl (12.4 g, 15.8 mmol) in 70 mL of acetonitrile is added at 2 ° C for 20 minutes. N-methylmorpholine is then added to obtain pH 7. The cooling is terminated and the mixture is allowed to warm to room temperature, then stirring is continued for a total of 3 hours. The turbid reaction mixture is filtered (slow) and the white residue is dried on the filter overnight to yield 10.8 g of FMOC-Glu (OBut) -Arg (PMC) -Val-Asp (Allyl) -OBzl. He The filtered product is concentrated to give a gum, which is absorbed in 30 mL of acetonitrile and left to stand for 48 hours, producing an additional 3.45 g of product. The total yield of FMOC-Glu (OtBu) -Arg (PMC) -f Val-Asp (OAIIyl) -OBzl is 14.3 g. (Mcaic 1192; M + 10bSvd 1192.4) 5 Synthesis of Glu (OtBu) -Arq (PMC) -Val-Asp (OAIyll) -OBzl To a solution of FMOC-Glu (OBut) -Arg (PMC) -Val-Asp ( OAII) - OBzl (12.3 g, 10.3 mmol) in 105 mL of dichloromethane is added piperidine (3.5 g, 41.3 mmol) at room temperature. The mixture is stirred for 2 hours, then it is diluted with heptane, concentrated to half the volume, and left to stand in the freezer overnight. The solution is decanted from the oily residue, which is absorbed in dichloromethane. The solution is washed with 0.5M aqueous citric acid, water, and brine, dried (magnesium sulfate), filtered, and concentrated. The crude solid product is crushed twice with heptane and filtered to give 9.0 g of Giu (OtBu) - (L) Arg (PMC) -Val-Asp (OAIIyl) - OBzl as a white powder. (Mcaic 970.2; M + 1"bSvd 971).

Synthesis of FMOC-LvsfAllocl-GlufOtBuVArqfPMO-Val-AspfOAIIvIVOBzl 20 A suspension of FMOC-Lys (Alloc) (4.2 g, 9.3 mmoles) and benzotriazole-loxi-tris [pyrrolidino] -phosphono hexafluoroacetate (PyBOP, 4.8 g 9.3 mmoles) in 69 mL of acetonitrile at 5 ° C is treated with N-methylmorpholine (0.94 g, 9.3 mmol), resulting in the formation of a solution which is stirred at 5 ° C for 30 minutes. A suspension of Glu (OtBu) -Arg (PMC) -Val-Asp (Allyl) -OBzl (9 g, 9.3 mmoles) in 52 mL of acetonitrile is added, causing the reaction mixture to turn turbid white. N-f Methylmorpholine is added until the reaction mixture is at pH 7, then the mixture is allowed to warm to room temperature and is stirred for 1.33 hours. The mixture is filtered and the solid residue is washed with acetonitrile (2 x 40 mL) and dried overnight under vacuum with a nitrogen purge to give 12.8 g of an off-white crusted solid crushed product that is comminuted with 200 g. mL of methyl butyl methyl ether, collected by filtration and dried under vacuum to yield 9.7 g of FMOC-Lys (Alloc) -Glu (OtBu) -Arg (PMC) -Val-Asp (OAIIyl) -OBzl. (Mcaic 1404.7; M + 16bsvd 1406) b. Removal of Alloc v Allyl from Lvs18 v Asp22: Synthesis of FMOC-15 Lvs-Glu (OtBu) -Arq (PMC) -Val-Asp-OBzl A 3-neck flask of 1 L equipped with mechanical agitator and nitrogen flux is charged at room temperature with 167 mL of dichloromethane and 1.37 g (1.18 mmol, 0.2 eq.) of Pd (PPh3). Nitrogen is bubbled into the orange solution for 2 minutes, then a solution of 8.3 g (5.91 mmoles, 1.0 eq.) Of FMOC-Lys (Alloc) -Glu (OtBu) -Arg (PMC) -Val- is added. Asp (OAIIyl) -OBzl in 66.4 mL of dimethylformamide, followed by 13.3 g, (124 mmol, 21 eq.) Of methylaniline. The solution is stirred at room temperature for 1 hour. The mixture is diluted with 700 mL of ether methyl-t-butyl that causes a solid product to form. The suspension is stirred for 1 hour and filtered. The solid residue is triturated with 200 mL of methyl-t-butyl ether, filtered and dried to obtain 5.0 g of a product? Solid pale yellow color. The filtrate produces 1.1 g of a second production as a pale yellow solid product for a total yield of 6.1 g of FMOC-Lys-Glu (OtBu) -Arg (PMC) -Val-Asp-OBzl (Mcaie 1280.6; M + 1ßbsvd 1281). c. Formation of Lactama Bridge of Lvs18-Asp22: Synthesis of H? FMOC-cycle (Lvs-Asp) Lvs-GlufOtBu) -Arq (PMC) -Val-Asp-OBzl 6.0 g (4.69 mmol) of FMOC-Lys-Glu (OtBu) are dissolved in a 3-neck flask of 1 L - Arg (PMC) -Val-Asp (OAIIyl) -OBzl in 300 mL of dimethylformamide at room temperature under nitrogen and 3.44 g, (6.61 mmol, 1.41 eq.) Of benzotriazolyloxytris [pyrrolidinyl] -15-phosphonium hexafluoroacetate ( PyBOP), 0.89 g, (6.57 mmol, 1.40 eq.) Of hydroxybenzotriazole hydrate, and 1.73 g, (13.4 mmol, 2.85 eq.) Of diisopropylethylamine are added. The orange solution is stirred for 1 hour at room temperature, then diluted with 300 mL of water resulting in the formation of an oily residue. The cloudy solution (containing mostly impurities) decanted. The oily residue is stirred overnight with 100 mL of water, yielding a crusty solid product which is collected by filtration, giving 4.2 g of a pale yellow solid product. The filtered product produces 1.0 g of a second production as a yellow solid product, for a total yield of 5.2 g of FMOC-cycle (Lys-Asp) Lys-Glu (OtBu) -Arg (PMC) -Val-Asp-OBzl. (M ^ c 1262 M + or 1262.5). d. Benzyl Ester Removal: Synthesis of FMOC-Cyclo (Lvs-Asp) Lys-Glu (OtBu) -Arq (PMC) -Val-Asp-OH A solution of 5.9 g (4.7 mmoles) of FMOC-Cyclo (Lys-Asp) Lys- Glu-Arg-Val-Asp-OBzl in 25 mL of dimethylformamide is treated with 5.9 carbon and filtered, and then treated with 0.59 g of 10% Pd / C and placed under f 10 3.51 kg / cm2 of hydrogen in a Parr apparatus. After 1.5 hours, the mixture is filtered, treated again with charcoal, refiltered and reloaded with 0.59 g of 10% Pd / C and exposed to 3.51 kg / cm2 of hydrogen for an additional 2 hours. . The mixture is filtered and the filtrate is diluted with 50 mL of water and extracted with three portions of 200 mL of sodium chloride. methylene. The combined methylene chloride extracts are concentrated to yield 1.2 g of FMOC-cyclo (Lys-Asp) Lys-Glu (OtBu) -Arg (PMC) -Val-f Asp-OH as an oil. (Mcaic 1171; M + 10bsvd 1172.3). 3. Purification 20 A 400 mg sample of FMOC-Cyclo (Lys-Asp) Lys-Glu (OtBu) -Org (PMC) -Val-Asp-OH is dissolved in 10 mL 1: 1 trifluoroethanol / isopropanol.

The sample is injected in two 5 mL portions into a preparation HPLC column (Microsorb MV C18, 300Ang, 8u, 41.4 X 250 mm) equilibrated with 55% B (A = 0.1% vl / vl aqueous trifluoroacetic acid, B = 75% v / v acetonitrile / 25% isopropanol with 0.1% trifluoroacetic acid). The load is eluted at 70 mL / minute in a gradient from 55% B to 65% B for 30 minutes, 65% B to 95% B for 1 minute, followed by washing of isocratic column with 95% B for 10 minutes. The effluent is monitored using 210nM UV rays. The fractions with a purity of 93.5A% or higher are deposited and concentrated by rotary evaporation. Lyophilization of the residue yields 200 mg of FMOC-cyclo (Lys-Asp) Lys-Glu-Arg-Val-Asp-OH. (Mcaic 1171; M + 10bsvd 1172.6).

Step 3 Resin of Fmoc-Lys-Glu-Arg-Val-Asp-Trp (Boc) -Leu-Arg (Pmc) - Lys (Boc) -Leu-Leu-Gln (Trt) -Asp (OtBu) -Val-NH FMOC resin- [TrP23 (BOC), Arg25 (PMC), Lys26 (BOC), Leu27, Gln29 (Trt)] HPTH (23-31) -chlorotryl (1 g, 0.185 mmol) was swollen in 10 mL of dimethylformamide with stirring for 30 minutes, then drained and treated with 10 mL of 20% piperidine / dimethylformamide (v / v) with agitation of 30 minutes. The resin was drained and washed with 3 portions of 20 mL of dimethylformamide. A solution of 240 mg (0.205 mmole 1.1 eq.) Of FMOC-cyclo (Lys18-Asp22) Lys-Glu (OtBu) -Arg (PMC) -Val-Asp-OH, 312 mg (0.6 mmoles 3 eq.) Of hexafluoroacetate of benzotriazolyloxy-tris [pyrrolidino] -phosphonium (PyBOP), and 156 mg (1.2 mmol, 6 eq) of diisopropylethylamine in 10 mL of dimethylformamide was added and the mixture was stirred for 16 hours. The resin was drained and washed with three 20 mL portions of dimethylformamide (TNBS positive) and re-treated with FMOC-cycle (Lys18-Asp2) Lys-Glu (OtBu) -Arg (PMC) -Val-Asp-OH / PyBOP / iPr2NEt a second time for 16 hours, then the resin was drained and washed with three portions of 10 mL of dimethylformamide (TNBS: negative). An aliquot of the title peptide was cleaved from the resin as described in step 2, Method A using Reagent K, to produce the side chain deprotected peptide fragment. (LC-MS: Mcaic2001; Mobsvd2000.5). The rest was used in step 4.

Step 4 Resin of Fmoc-Lys (Boc) -H¡s (Trt) -Leu-Asn (Trt) -Ser (OtBu) -Lys-Glu (OtBu) -Arg (PMC) -Val-Aspl-Trp (Boc) -Leu -Arg (PMC) -Lys (Boc) -Leu-Leu-Gln (Trt) -Asp (OtBu) -Val-NH The desired resin-bound peptide was prepared by removing the protective Fmoc group of 1.38 g (0.244 mmol with titre base in Trp23) of the resin-bound peptide from step 3 according to step 1, followed by consecutive cyclization of FMOC removal and coupling with 0.5 mmoles of each of FMOC-Ser (OtBu) -OH, FMOC-Asn (Trt) -OH, FMOC-Leu-OH, FMOC-His (Trt) -OH, and FMOC-Lys (BOC) -OH according to step 1, using 190 mg (0.5 mmol) of O-benzotriazole hexafluorophosphate -1-yl-N, N, N ', N'-tetramethyluronium (HBTU) and 2 mmoles) of N-methylmorpholine in 25 mL of dimethylformamide. The couplings were performed at room temperature for about 1 hour. The final resin peptide was washed with portions of 25 mL of dimethylformamide, tetrahydrofuran, and diethyl ether and dried under vacuum (1.93 g). An aliquot of the title peptide was cleaved from the resin as described below (Reagent K), to f produce the side-chain deprotected peptide fragment (LC-MS: Mcaic2579.4; MobSvd2580.7) of 25A% purity.

Step 5 Fmoc-Ala-Val-Ser (OtBu) -Glu (OtBu) -lle-Gln (Trt) -Leu-Nle-His (Trt) - Asn (Trt) -Leu-Gly-OH FMOC- [Ala1, Ser ^ OtBu), Glu4 (OtBu), Gln8 (Trt), His9 (Trt), f 10 Asn10 (Trt) Nle8] hPTH (1-12) The title peptide was prepared by coupling 2.65 g (5 mmoles) of Fmoc-Leu-OH at 5 g (3 mmoles) of Gly-2-chlorotrityl resin crosslinked to 1% of 0.6 meq / g with 3.9 g (7.5 mmol) of benzotriazolyloxytris [pyrrolidino] -phosphonium hexafluoroacetate (PyBOP), 1.94 g (15 mmol) of diisopropylethylamine in 50 mL of dimethylformamide / tetrahydrofuran (v / v) using stirring, for 0.5 hours. The resin was drained, then f washed with five portions of dimethylformamide / tetrahydrofuran and one portion of methanol (Ninhydrin: negative). The resin was washed once with dimethylformamide, then treated twice with 60 mL of piperidine % / dimethylformamide for 15 minutes each time, with three portions of 60 mL of dimethylformamide and a 60 mL portion of methanol between each treatment. After the second treatment, ninhydrin was highly positive. This cycle was repeated, using a piperidine treatment 0.5-hour dimethylformamide unique in each subsequent case, and using 2.5 molar equivalents of each of FMOC-Asn (Trt), FMOC-His (Trt), FMOC-Nle, FMOC-Leu, FMOC-Gln (Trt), FMOC- lle, FMOC, Glu (OtBu), FMOC-Ser (OtBu), FMOC-Val, and FMOC-Ala, together with 2.5 molar equivalents of each of PyBOP and diisopropylethylamine. The resin peptide was cleaved by stirring with 100 mL of acetic acid / trifluoroethylene / and dichloromethane (1: 1: 8) for 1 hour. The liquid was drained from the resin, and the resin was washed with 200 mL of dichloromethane; then the wash was combined with the main portion of the liquid. This was evaporated consecutively from hexane, ethyl acetate, and sodium propane. The residue was triturated with diethyl ether, then dried (overnight, vacuum) to yield 2.24 g of the title peptide (HPLC: 83% pure). The resin was re-treated under segmentation conditions a second time and the liquid was treated as after the first cleavage, to finally yield 2.75 g of a second production of the title peptide, for a total yield of 4.95 g (2.1 mmol, 70% resin yield). [LC-MS (Mcaic1514.8; M0bsvd 514.7] This material was used as obtained in step 6.

Step 6 Fmoc-Ala-Val-Ser (OtBu) -Glu (? TBu) -lle-Gln (Trt) -Leu-Nle-His (Trt) -Asn (Trt) -Leu-Gly-Lys (Boc) resin -His (Trt) -Leu-Asn (Trt) -Ser (? TBu) -Lys-Glu (OtBu) -Arg (PMC) -Val-Asp- Trp (Boc) -Leu-Arg (PMC) -Lys (Boc) ) -Leu-Leu-Gln (Trt) -Asp (? TBu) -Val-NH The peptide prepared in step 5 (900 mg, 0.114 mmol) was swollen in 15 mL of dimethylformamide for 40 minutes with shaking, then the resin was drained and treated with 15 mL of 20% piperidine / dimethylformamide (v / v) during 45 minutes. The resin was drained and washed with three 30 mL portions of dimethylformamide (resin TNBS: positive). To this were added 15 mL of dimethylformamide, 775 mg, (0.33 mmoles, 2.7 molar equivalents based on titration in the position 23) of FMOC- [Ala1, Ser ^ OtBu), Glu4 (OtBu), Gln6 (Trt), His9 (Trt), Asn10 (Trt) Nleß] hPTH (1-12) (from step 4), 151 mg (0.29 mmol, 0.21 molar equivalents) of benzotriazolyloxytris [pyrrolidino] -phosphonium hexafluoroacetate (PyBOP), 39 mg (0.25 moles) of Hydroxybenzotriazole hydrated and 75 mg (0.58 mmol) of diisopropylethylamine, and the mixture was stirred for 3 days.

The resin was drained, washed with three 30 mL portions of dimethylformamide, (TNBS, mixture of positive and negative spheres), then treated with 15 mL of 20% piperidine / dimethylformamide (v / v), drained, and washed with three 15 mL portions of each of dimethylformamide, tetrahydrofuran, and diethyl ether, then dried and deprotected and segmented from the resin according to step 7.

Step 7 Ala-Val-Ser-Glu-lle-GIn-Leu-Nle-His-Asn-Leu-Gly-Lys-His-Leu-Asn- Ser-Lys-Glu-Arg-Val-Asp-Trp-Leu-Arg-Lys-Leu-Leu-GIn-Asp-Val-NH 2 The resin peptide from step 6 was treated to remove FMOC in Ala1 according to Method A of step 1, then segmented and de-protected simultaneously in a mixture of 33/2/2/1/2 of trifluoroacetic acid, phenol, thioanisole, ethanedithiol, and water (total dilution of Reagent K mL / g resin peptide) for 3 hours. The resin was drained and the product was precipitated from the filtrate by adding the reaction mixture to cold t-butylmethyl ether (50 mL / g of resin peptide). The mixture was allowed to stand for 15 minutes, then centrifuged (2000 rpm, 5 minutes). The liquid was decanted and the remaining solid product was washed with diethyl ether, the supernatant of the mixture was separated by centrifugation and the supernatant was removed. The resulting solid product is stirred in 0.1% aqueous trifluoroacetic acid (6 mL / g resin peptide) for 1 hour to ensure complete removal of acid sensitive groups. The solution is lyophilized to produce 414 mg of the title peptide (37% pure, 26.3% pure in w / w, yield of 26.3% of Tro23) as a white solid product.

(LC-MS: Mra? G 3632.0; M + 10bsvd3633.4).

Step 8: Purification One gram of the material prepared above dissolves in 10 mL of water and the solution is injected in a column of preparation HPLC 5.08 x 25 cm (10 microns, 120A or 300A) pre-equilibrated with 75/5/20 of 0.1% aqueous trifluoroacetic acid (A) / trifluoroacetic acid 0. 1% in acetonitrile (B) / isopropanol (C). The column elutes at 115 L per minute with a gradient 75A / 5B / 20C at 60A / 20B / 20C for 30 minutes, then washed with 30% A, 60% B, 20% C and rebalanced with 75A / 5B / 20C. The effluent is monitored using UV detection at 280 nm. Fractions containing the title peptide of area purity percentage 5 of about 75 are combined and concentrated in a rotary evaporator (~ 35 ° C). The concentrated aqueous solution is pumped into the same preparation column and run again through the same 30 minute gradient. Fractions containing the title peptide at 90A% or higher percent area purity are combined and concentrated in a rotary evaporator at (~ 35 ° C). Then the concentrated solution is pumped into the preparation column which had been pre-equilibrated with 75% aqueous ammonium acetate 0.1 M (adjusted to pH 6 with acetic acid) (D) / 15% acetonitrile (E) / isopropanol to 10% (C). The column is eluted at 115 mL / minute with 75D / 15E / 10C at 60D / 30E / 10C for 30 minutes, then washed with 15 30D / 60E / 20C. Fractions containing the title peptide of 95A% or higher percentage of area purity are combined and lyophilized, providing a purified product such as the solid acetate salt (1 g of crude gives approximately 100-200 mg @ approximately 95A% purity) . At each stage, the fractions are analyzed for content and purity using analytical HPLC on a C18 column (4.6 mm x 25 cm, 5 microns 120A or 300A) in a gradient of 25-40% B (A = trifluoroacetic acid) 0.1% / water, B = 0.1% trifluoroacetic acid / acetonitrile) for 30 minutes using a flow of 1 mL / minute and monitoring with UV at 220 nm.

LIST OF SEQUENCES < 110 > Sledeski, Adam W. Mencel, James J. < 120 > PROCEDURE FOR THE PREPARATION OF CYCLIC PEPTIDES UNITED TO RESIN < 130 > A3113A-WO < 140 > < 141 > < 150 > 60/081, 897 < 151 > 1998-04-15 < 160 > 46 < 170 > Patentln Ver. 2.1 < 210 > 1 < 211 > 31 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > PEPTIDE < 222 > (18) .. (22) < 223 > "Synthetic peptide: The side chains of Lys at position 18 and Asp in position 22 are linked by an amide bond " < 220 > < 221 > MOD_RES < 222 > (8) < 223 > NIe < 220 > < 221 > PEPTIDE < 222 > (31) < 223 > "This C-terminal amino acid is an amide, that is, CONH2" < 400 > 1 'Ala Val Ser Glu Lie Gln Leu Xaa His Asn Leu sly Lys His Leu Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val 20 25 30 < 210 > 2 < 211 > 31 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > MOD_RES < 222 > (8) < 223 > NIe < 220 > < 221 > PEPTIDE < 222 > (18) .. (22) < 223 > "Synthetic peptide: The side chains of Lys in position 18 and Asp in position 22 are linked by an amide bond" < 220 > < 221 > PEPTIDE < 222 > (31) < 223 > "This C-terminal amino acid is an amide, that is, CONH2" < 400 > 2 Ala Ala Ser Glu He Gln Leu Xaa His Asn Leu Gly Lys His Leu Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val 20 25 30 < 210 > 3 < 211 > 31 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > MOD_RES < 222 > (8) < 223 > NIe < 220 > < 221 > PEPTIDE < 222 > (18) .. (22) < 223 > "Synthetic peptide: The side chains of Lys in position 18 and Asp in position 22 are linked by an amide bond" < 220 > < 221 > PEPTIDO < 222 > (31) < 223 > "This C-terminal amino acid is an amide, that is, CONH2" < 400 > 3 Wing Val Wing Glu He Gln Leu Xaa His Asn Leu Gly Lys His Leu Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val 20 25 30 < 210 > 4 < 211 > 31 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > MOD_RES < 222 > (8) < 223 > NIe < 220 > < 221 > PEPTIDE < 222 > (18) .. (22) < 223 > "Synthetic peptide: The side chains of Lys at position 18 and Asp in position 22 are linked by an amide bond " < 220 > < 221 > PEPTIDE < 222 > (31) < 223 > "This C-terminal amino acid is an amide, that is, CONH2" < 400 > 4 Wing Val Wing Wing He Gln Leu Xaa His Asn Leu Gly Lys His Leu Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val • 10 < 210 > 5 < 211 > 31 < 212 > PRT < 213 > Artificial Sequence 15 < 220 > < 221 > MOD_RES < 222 > (8) < 223 > NIe 20 < 220 > < 221 > PEPTIDE < 222 > (18) .. (22) < 223 > "Synthetic peptide: The side chains of Lys at position 18 and Asp in position 22 are linked by an amide bond " < 220 > < 221 > PEPTIDE < 222 > (31) < 223 > "This C-terminal amino acid is an amide, that is, CONH2" < 400 > 5 Wing Val Ser Glu Wing Gln Leu Xaa His Asn Leu Gly Lys His Leu Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val 20 25 30 < 210 > 6 < 211 > 31 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > MOD_RES < 222 > (8) < 223 > NIe < 220 > < 221 > PEPTIDE < 222 > (18) .. (22)) < 223 > "Synthetic peptide: The side chains of Lys in position 18 and 5 Asp in position 22 are linked by an amide bond" < 220 > < 221 > PEPTIDE < 222 > (31) H? < 223 > "This C-terminal amino acid is an amide, that is, CONH2" < 400 > 6 Wing Val Ser Glu He Wing Leu Xaa His Asn Leu Gly Lys His Leu Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val 20 25 30 15 < 210 > 7 < 211 > 31 < 212 > PRT 20 < 213 > Artificial Sequence < 220 > < 221 > MOD RES < 222 > (8) < 223 > NIe ) < 220 > 5 < 221 > PEPTIDE < 222 > (18) .. (22) < 223 > "Synthetic peptide: The side chains of Lys at position 18 and Asp in position 22 are linked by an amide bond " (10 <220> <221> PEPTIDE <222> (31) <223> "This C-terminal amino acid is an amide, that is, CONH2" < 400 > 7 Ala Val Ser Glu He Gln Ala Xaa His Asn Leu Gly Lys His Leu Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val 20 25 30 < 210 > 8 < 211 > 31 < 212 > PRT < 213 > (Artificial Sequence) < 220 > < 221 > PEPTIDE < 222 > (18) . (22) m < 223 > "Synthetic peptide: The side chains of Lys in position 18 and Asp 5 in position 22 are linked by an amide bond" < 220 > < 221 > PEPTIDE < 222 > (31) 110 < 223 > "This C-terminal amino acid is an amide, that is, CONH2." < 400 > 8 Wing Val Sef Glu He Gln Leu Wing His Asn Leu Gly Lys His Leu Asn 1 5. 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val 20 25 30 15 < 210 > 9 < 211 > 31 < 212 > PRT 20 < 213 > (Artificial Sequence) < 220 > < 221 > MOD_RES < 222 > (8) < 223 > NIe < 220 > ) < 221 > PEPTIDE 5 < 222 > (18) . (22) < 223 > "Synthetic peptide: The side chains of Lys in position 18 and Asp in position 22 are linked by an amide bond." < 220 > , 10 < 221 > PEPTIDE < 222 > (31) < 223 > "This C-terminal amino acid is an amide, that is, CONH2." < 400 > 9 Wing Val Ser Glu He Gln Leu Xaa Hzs Wing Leu Gly Lys His Leu Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val 20 25 30 < 210 > 10 20 < 211 > 31 < 212 > PRT < 213 > (Artificial Sequence) < 220 > < 221 > MOD_RES < 222 > (8) < 223 > NIe < 220 > < 221 > PEPTIDE < 222 > (18) . (22) < 223 > "Synthetic peptide: The side chains of Lys in position 18 and Asp in position 22 are linked by an amide bond." (10 <220> <221> PEPTIDE <222> (31) <223> "This C-terminal amino acid is an amide, ie CONH2." < 400 > 10 Wing Val Ser Glu He Gln Leu Xaa His Asn Wing Gly Lys His Leu Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val 20 25 30 < 210 > 11 < 211 > 31 < 212 > PRT < 213 > (Artificial Sequence) < 220 > < 221 > M0D_RES < 222 > (8)) < 223 > NIe 5 < 220 > < 221 > PEPTIDE < 222 > (18) .. (22) < 223 > "Synthetic peptide: The side chains of Lys at position 18 and Asp ^ 10 at position 22 are linked by an amide bond." < 220 > < 221 > PEPTIDE < 222 > (31) 15 < 223 > "This C-terminal amino acid is an amide, that is, CONH2." Ala Val Ser Glu He Gln Leu Xaa His Asn Leu Ala Lys His Leu Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val twenty < 210 > 12 < 211 > 31 < 212 > PRT < 213 > (Artificial Sequence) < 220 > • < 221 > M0D_RES 5 < 222 > (8) < 223 > NIe < 220 > < 221 > PEPTIDE) 10 < 222 > (18) . (22) < 223 > "Synthetic peptide: The side chains of Lys in position 18 and Asp in position 22 are linked by an amide bond." < 220 > 15 < 221 > PEPTIDE < 222 > (31) i < 223 > "This C-terminal amino acid is an amide, that is, CONH2." < 400 > 12 0 A? A Va? Ser G u ?? e Q n Leu? Aa HÍS Asn Leu Gly Ala His Leu Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val 20 25 30 < 210 > 13 < 211 > 31 < 212 > PRT < 213 > (Artificial Sequence) < 220 > < 221 > MOD_RES < 222 > (8) < 223 > NIe < 220 > < 221 > PEPTIDE < 222 > (18) . (22) < 223 > "Synthetic peptide: The side chains of Lys in position 18 and Asp in position 22 are linked by an amide bond." < 220 > < 221 > PEPTIDE < 222 > (31) < 223 > "This C-terminal amino acid is an amide, that is, CONH2." < 400 > 13 Ala Val Ser Glu He Gln Leu Xaa His Asn Leu Gly Lys Ala Leu Asn 1 5? O? 5 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val < 210 > 14 < 211 > 31 < 212 > PRT < 213 > (Artificial Sequence) 5 < 220 > < 221 > MOD_RES < 222 > (8) < 223 > NIe or < 220 > < 221 > PEPTIDE < 222 > (18) .. (22) < 223 > "Synthetic peptide: The side chains of Lys at position 18 and Asp 5 at position 22 are linked by an amide bond." < 220 > < 221 > PEPTIDE < 222 > (31) 0 < 223 > "This C-terminal amino acid is an amide, that is, CONH2." < 400 > 14 Ala Val Ser Glu He Gln Leu Xaa His Asn Leu Gly Lys His Wing Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val 20 25 30 • 5 < 210 > 15 < 211 > 31 < 212 > PRT < 213 > (Artificial Sequence) • 10 < 220 > < 221 > MOD_RES < 222 > (8) < 223 > NIe 15 < 220 > < 221 > PEPTIDE < 222 > (18) .. (22) < 223 > "Synthetic peptide: The side chains of Lys in position 18 and Asp 20 in position 22 are linked by an amide bond." < 220 > < 221 > PEPTIDE < 222 > (31) < 223 > "This C-terminal amino acid is an amide, that is, CONH2." < 400 > 15 Ala Val Ser Glu He Gln Leu Xaa His Asn Leu Gly Lys His Leu Ala 1 5 10 15 • Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val 20 25 30 < 210 > 16 < 211 > 31 > 10 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > MOD_RES 15 < 222 > (8) < 223 > NIe < 220 > < 221 > PEPTIDE 20 < 222 > (13) .. (17) < 223 > "Synthetic peptide: The side chains of Lys at position 13 and Asp at position 17 are linked by an amide bond." < 220 > < 221 > PEPTIDE < 222 > (18) .. (22) f < 223 > "Synthetic peptide: The side chains of Lys at position 18 and Asp 5 at position 22 are linked by an amide bond." < 220 > < 221 > PEPTIDE < 222 > (31) f 10 < 223 > "This C-terminal amino acid is an amide, that is, CONH2." < 400 > 16 Ala Val Ser Glu He Gln Leu Xaa His Asn Leu Gly Lys His Leu Asn 1 i 5 10 15 Asp Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val fifteen < 210 > 17 < 211 > 31 < 212 > PRT 20 < 213 > Artificial Sequence < 220 > < 221 > MOD RES < 222 > (8) < 223 > NLE f < 220 > 5 < 221 > PEPTIDE < 222 > (18) .. (22) < 223 > "Synthetic peptide: The side chains of Lys in position 18 and Asp in position 22 are linked by an amide bond." f l O < 220 > < 221 > PEPTIDE < 222 > (26) .. (30) < 223 > "Synthetic peptide: The side chains of Lys at position 26 and Asp at position 30 are linked by an amide bond." < 220 > < 221 > PEPTIDE < 222 > (31) < 223 > "This C-terminal amino acid is an amide, that is, CONH2." < 400 > 17 Ala Val Ser Gl? lie Gln Leu Xaa His Asn Leu Gly Lys His Leu Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val 20 25 30 < 210 > 18 < 211 > 34 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > MOD_RES < 222 > (8) < 223 > NIe < 220 > < 221 > PEPTIDE < 222 > (18) .. (22) < 223 > "Synthetic peptide: The side chains of Lys in position 18 and Asp in position 22 are linked by an amide bond." < 220 > < 221 > PEPTIDE < 222 > (34) < 223 > "This C-terminal amino acid is an amide, that is, CONH2." < 400 > 18 Ala Val Ser Glu He Gln Leu Xaa His Asn Leu Gly Lys His Leu Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val His 20 25 30 < 210 > 19 < 211 > 34 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > MOD_RES < 222 > (8) < 223 > NIe < 220 > < 221 > PEPTIDE < 222 > (13) .. (17) < 223 > "Synthetic peptide: The side chains of Lys at position 13 and Asp at position 17 are linked by an amide bond." < 220 > < 221 > PEPTIDO < 222 > (18) .. (22) < 223 > "Synthetic peptide: The side chains of Lys in position 18 and Asp in position 22 are linked by an amide bond." < 220 > < 221 > PEPTIDE < 222 > (34) < 223 > "This C-terminal amino acid is an amide, that is, CONH2." < 400 > 19 Ala Val Ser Glu He Gln Leu Xaa His Asn Leu Gly Lys His Leu Asn 1 5? O is Asp Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val His 20 25 30 < 210 > 20 < 211 > 34 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > PEPTIDE < 222 > (18) .. (22) < 223 > "Synthetic peptide: The side chains of Lys in position 18 and Asp in position 22 they are linked by an amide bond. < 220 > < 221 > PEPTIDE < 222 > (34) < 223 > "This C-terminal amino acid is an amide, that is, CONH2." < 400 > 20 Wing Val Ser Glu His Gln Leu Leu His Asp Lys Gly Lys Ser He Gln 1 5 10 15 Asp Lys Arg Arg Arg Asp Phe Leu His His Leu He Wing Glu He His 20 25 30 < 210 > 21 < 211 > 34 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > PEPTIDE < 222 > (18) .. (22) < 223 > "Synthetic peptide: The side chains of Lys in position 18 and Asp in position 22 are linked by an amide bond." < 220 > < 221 > PEPTIDE < 222 > (34) < 223 > "This C-terminal amino acid is an amide, that is, CONH2." < 400 > 21 Ala Val Ser Glu His Gln Leu Leu His Asp Lys Gly Lys Ser He Gln 1 5 10 15 Asp Lys Arg Arg Arg Asp Leu Leu Glu Lys Leu Leu Glu Lys Leu His. 20 25 30 < 210 > 22 < 211 > 28 < 212 > PRT < 213 > Artificial Sequence 15 < 220 > I < 221 > PEPTIDE < 222 > (12) .. (16) < 223 > "Synthetic peptide: The side chains of Lys at position 12 and Asp 20 at position 16 are linked by an amide bond". < 220 > < 221 > PEPTIDO < 222 > (28) < 223 > "This C-terminal amino acid is an amide, that is, CONH2" < 220 > < 221 > MOD_RES < 222 > (2) < 223 > NIe < 400 > 22 Leu Xaa His Asn Leu Gly Lys His Leu Asn Ser Lys Glu Arg Val Asp 1 5 10 15 Trp Leu Arg Lys Leu Leu Gin Asp Val His Asn Phe 20 25 < 210 > 23 < 211 > 28 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > PEPTIDE < 222 > (12) .. (16) < 223 > "Synthetic peptide: The side chains of Lys in position 12 and Asp in the position of 16 are linked by an amide bond". < 220 > < 221 > PEPTIDE < 222 > (28) < 223 > "This C-thermal amino acid is an amide, that is, CONH2" < 400 > 23 Leu Leu His Asp Lys Gly Lys Ser He Gln Asp Lys Arg Arg Arg Asp 1, 5 10 15 Phe Leu His His Leu He Wing Glu He His Thr Wing 20 25 < 210 > 24 < 211 > 28 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > MOD_RES < 222 > (2) < 223 > NIe < 220 > < 221 > PEPTIDE < 222 > (7) .. (11) < 223 > "Synthetic peptide: The side chains of Lys in the 7 position and Asp in the position of 11 are linked by an amide bond". f < 220 > 5 < 221 > PEPTIDE < 222 > (12) . (16) < 223 > "Synthetic peptide: The side chains of Lys in position 12 and Asp in position of 16 are linked by amide bonds". f lO < 220 > < 221 > PEPTIDE < 222 > (28) < 223 > "This C-terminal amino acid is an amide, that is, CONH2." < 400 > 24 Leu Xaa His Asn Leu Gly Lys His Leu Asn Asp Lys Glu Arg Val Asp 1 5 10 15 f Trp Leu Arg Lys Leu Leu Gln Asp Val His Asn Phe ^ 20 25 < 210 > 25 < 211 > 28 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > MOD_RES < 222 > (2) < 223 > NIe < 220 > < 221 > PEPTIDE < 222 > (12) . (16) < 223 > "Synthetic peptide: The side chains of Lys in position 12 and Asp in the position of 16 are linked by an amide bond". < 220 > < 221 > PEPTIDE < 222 > (twenty) . . (24) < 223 > "Synthetic peptide: The side chains of Lys at position 20 and Asp at position 24 are linked by an amide bond". < 220 > < 221 > PEPTIDE < 222 > (28) < 223 > "This C-terminal amino acid is an amide, that is, CONH2." < 400 > 25 Leu xaa His Asn Leu Gly Lys His Leu Asn Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu leu Gln Asp Val His Asn Phe < 210 > 26 < 211 > 34 < 212 > PRT < 213 > Artificial Sequence < 400 > 26 Ser Val Ser Glu He Gln Leu Met His Asn Leu Gly Lys His Leu Asn 1 5? O 15 Ser Met Glu Arg Val Glu Trp Leu Arg Lys Lys Leu Gln Asp Val His 20 25 30 < 210 > 27 < 211 > 34 < 212 > PRT < 213 > Artificial Sequence < 400 > 27 Wing Val Ser Glu His Gln Leu Leu His Asp Lys Gly Lys Ser He Gln 1 5 10 15 Asp Leu Arg Arg Arg Phe Phe Leu His His Leu He Wing Glu He His 20 25 30 < 210 > 28 < 211 > 9 < 212 > PRT < 213 > Artificial Sequence • 10 < 220 > < 221 > PEPTIDE < 222 > (1) < 223 > FMOC-Trp (BOC) 15 < 220 > < 221 > PEPTIDE < 222 > (3) < 223 > product = Arg (PMC) 20 < 220 > < 221 > PEPTIDE < 222 > (4) < 223 > Product = Lys (BOC) < 220 > f < 221 > PEPTIDE 5 < 222 > (7) < 223 > product = Gln (Trt) < 220 > < 221 > PEPTIDO f lO < 222 > (8) < 223 > product = Asp (OtBu) < 220 > < 221 > PEPTIDO 15 < 222 > (9) < 223 > Synthetic peptide Other- "This C-terminal amino acid is linked by faith an amide bond to the resin of Rink Amida MBHA." < 400 > 28 20 Trp Leu Arg Lys Leu Leu Gln Asp Val 1 5 < 210 > 29 < 211 > 14 < 212 > PRT < 213 > Artificial Sequence f < 220 > 5 < 221 > PEPTIDE < 222 > (1) < 223 > FMOC-Lys < 220 > lO < 221 > PEPTIDE < 222 > (1) .. (5) < 223 > "Synthetic peptide: The side chains of Lys in position 1 and Asp in the position of 5 are linked by an amide bond". < 220 > < 221 > PEPTIDE f < 222 > (2) < 223 > Glu (OtBu) < 220 > < 221 > PEPTIDE < 222 > (3) < 223 > Arg (PMC) < 220 > < 221 > PEPTIDE < 222 > (1) .. (5) I < 223 > "Synthetic peptide: The side chains of Lys in position 1 and Asp 5 in position 5 are linked by an amide bond." < 220 > < 221 > PEPTIDE < 222 > (6) io < 223 > Trp (BOC) < 220 > < 221 > PEPTIDE < 222 > (8) 15 < 223 > Arg (PMC) < 220 > < 221 > PEPTIDE < 222 > (9) 20 < 223 > Lys (BOC) < 220 > < 221 > PEPTIDO < 222 > (12) < 223 > Gln (Trt) I < 220 > 5 < 221 > PEPTIDE < 222 > (13) < 223 > Asp (OtBu) < 220 > ^ 10 < 221 > PEPTIDE < 222 > (14) < 223 > "This C-terminal amino acid is linked by an amide bond to the Rink Amida MBHA resin." < 400 > 29 Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val 5? < 210 > 30 < 211 > 19 20 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > PEPTlDO < 222 > (1) < 223 > FMOC-Lys (BOC) < 220 > < 221 > PEPTIDE < 222 > (2) < 223 > His (Trt) < 220 > < 221 > PEPTIDE < 222 > (4) < 223 > Asn (Trt) < 220 > < 221 > PEPTIDE < 222 > (5) < 223 > Ser (OtBu) < 220 > < 221 > PEPTIDE < 222 > (6) .. (10) < 223 > "Synthetic peptide: The side chains of Lys in position 6 and Asp in position 10 they are linked by an amide bond. " < 220 > < 221 > PEPTIDE < 222 > (7) < 223 > Glu (OtBu) < 220 > < 221 > PEPTIDE < 222 > (8) < 223 > Arg (PMC) < 220 > < 221 > PEPTIDE < 222 > (6) .. (10) < 223 > "Synthetic peptide: The side chains of Lys in position 6 and Asp in position 10 are linked by an amide bond." < 220 > < 221 > PEPTIDE < 222 > (11) < 223 > Trp (BOC) < 220 > < 221 > PEPTIDE < 222 > (13) < 223 > Arg (PMC) < 220 > < 221 > PEPTIDE < 222 > (14) < 223 > Lys (BOC) < 220 > < 221 > PEPTIDE < 222 > (17) < 223 > Gln (Trt) < 220 > < 221 > PEPTIDE < 222 > (18) < 223 > Asp (OtBu) < 220 > < 221 > PEPTIDE < 222 > (6) .. (10) < 223 > "Synthetic peptide: The side chains of Lys in position 6 and Asp in position 10 are linked by an amide bond." < 400 > 30 Lys His Leu Asn Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu 1 5 10 15 < 210 > 31 < 211 > 12 < 213 > Artificial Sequence < 220 > < 221 > PEPTIDO 15 < 222 > (1) < 223 > FMOC-Ala • < 220 > < 221 > PEPTIDE 20 < 222 > (3) < 223 > Ser (OtBu) < 220 > < 221 > PEPTIDO < 222 > (4) < 223 > Glu (OtBu) < 220 > < 221 > PEPTIDO < 222 > (6) < 223 > Gln (Trt) < 220 > < 221 > PEPTIDO < 222 > (9) < 223 > His (Trt) < 220 > < 221 > PEPTIDO < 222 > (10) < 223 > Asn (Trt) < 220 > < 221 > MOD_RES < 222 > (8) < 223 > NIe < 400 > 31 Ala val Ser Glu He Gln Leu Xaa His Asn Leu Gly | < 210 > 32 5 < 211 > 31 < 212 > PRT < 213 > Artificial Sequence < 220 > H? < 221 > PEPTIDE < 222 > (1) < 223 > FMOC-Ala < 220 > 15 < 221 > PEPTIDE < 222 > (3) < 223 > Ser (OtBu) < 220 > 20 < 221 > PEPTIDE < 222 > (4) < 223 > Glu (OtBu) < 220 > < 221 > PEPTIDE < 222 > (6) < 223 > Gln (Trt) < 220 > < 221 > PEPTIDE < 222 > (9) < 223 > His (Trt) < 220 > < 221 > PEPTIDE < 222 > (10) < 223 > Asn (Trt) < 220 > < 221 > PEPTIDE < 222 > (13) < 223 > Lys (BOC) < 220 > < 221 > PEPTIDO < 222 > (14) < 223 > His (Trt) < 220 > | < 221 > PEPTIDE 5 < 222 > (16) < 223 > Asn (Trt) < 220 > < 221 > PEPTIDE ^ 10 < 222 > (17) < 223 > Ser (OtBu) < 220 > < 221 > PEPTIDO 15 < 222 > (18) .. (22) < 223 > "Synthetic peptide: the side chains of Lys in position 18 and Asp in position 22 are linked by an amide bond" < 220 > 20 < 221 > PEPTIDE < 222 > (19) < 223 > Glu (OtBu) < 220 > < 221 > PEPTIDE < 222 > (20) f < 223 > Arg (PMC) 5 < 220 > < 221 > PEPTIDE < 222 > (22) . (30) < 223 > "Synthetic peptide: the side chains of Lys in position 13 and Asp flO in position 30 are linked by an amide bond" < 220 > < 221 > PEPTIDE < 222 > (23) 15 < 223 > Trp (BOC) f < 220 > < 221 > PEPTIDE < 222 > (25) 20 < 223 > Arg (PMC) < 220 > < 221 > PEPTIDO < 222 > (26) < 223 > Lys (BOC) ) < 220 > 5 < 221 > PEPTIDE < 222 > (29) < 223 > Gln (Trt) < 220 > (10 <221> PEPTIDE <222> (18) < 223 > Asp (OtBu) < 220 > 15 < 221 > PEPTIDE < 222 > (30) < 223 > "This C-terminal amino acid is linked by an amide bond to the Rink Amida MBHA resin." < 220 > < 221 > MOD-RES < 222 > (8) < 223 > X = Norleucine; NIe < 400 > 32 Ala Val Ser Glu He Gln Leu Xaa His Asn Leu Gly Lys His Leu Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val 20 25 30 < 210 > 33 < 211 > 2 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > PEPTIDE < 222 > (2) < 223 > Asp (OAIIyl) < 220 > < 221 > PEPTIDE < 222 > (1) < 223 > BOC-Val < 220 > < 221 > PEPTIDE < 222 > (2) < 223 > "This C-terminal amino acid is in the form of an ester with benzyl alcohol." < 400 > 33 Val Asp 1 < 210 > 34 < 211 > 2 < 212 > PRT < 223 > Artificial Sequence < 220 > < 221 > PEPTIDE < 222 > (1) < 223 > "The N-terminal is in the form of a hydrochloride salt." < 220 > < 221 > PEPTIDE < 222 > (2) < 223 > Asp (OAIIyl) < 220 > < 221 > PEPTIDO < 222 > (2) < 223 > "This C-terminal amino acid is in the form of an ester with benzyl alcohol." < 400 > 34 Val Asp > 210 > 35 < 211 > 3 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > PEPTIDE < 222 > (1) < 223 > FMOC-Arg (PMC) < 220 > < 221 > PEPTIDE < 222 > (3) < 223 > Asp (OAIIyl) < 220 > < 221 > PEPTIDE < 222 > (3) < 223 > "This C-terminal amino acid is in the form of an ester with benzyl alcohol." 5 < 400 > 35 Arg Val Asp 1 < 210 > 36? < 211 > 3 < 212 > PRT < 213 > Unknown < 220 > 5 < 221 > PEPTIDE < 222 > (1) < 223 > Arg (PMC) < 220 > 0 < 221 > PEPTIDE < 222 > (3) < 223 > Asp (OAIIyl) < 220 > < 221 > PEPTIDE < 222 > (3) < 223 > "This C-terminal amino acid is in the form of an ester with benzyl alcohol." < 400 > 36 Arg Val Asp 1 ? < 210 > 37 < 211 > 4 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > PEPTIDE < 222 > (1) < 223 > FMOC-Glu (OtBu) 0 < 220 > < 221 > PEPTIDE < 222 > (2) < 223 > Arg (PMC) < 220 > < 221 > PEPTIDE < 222 > (4) | < 223 > Asp (OAIIyl) 5 < 220 > < 221 > PEPTIDE < 222 > (4) < 223 > "This C-terminal amino acid is in the form of an ester with benzyl alcohol." < 400 > 37 Glu Arg Val Asp 1 < 210 > 38 < 211 > 4 f < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > PEPTIDE < 222 > (1) < 223 > Glu (OtBu) < 220 > < 221 > PEPTIDE < 222 > (2) < 223 > Arg (PMC) < 220 > < 221 > PEPTIDE < 222 > (4) < 223 > Asp (OAIIyl) < 220 > < 221 > PEPTIDE < 222 > (4) < 223 > "This C-terminal amino acid is in the form of an ester with benzyl alcohol." < 400 > 38 Glu Arg Val Asp 1 < 210 > 39 < 211 > 5 < 212 > PRT < 223 > Artificial Sequence < 220 > < 221 > PEPTIDO < 222 > (1) < 223 > FMOC-Lys (Alloc) < 220 > < 221 > PEPTIDE < 222 > (2) < 223 > Glu (OtBu) < 220 > < 221 > PEPTIDE < 222 > (3) < 223 > Arg (PMC) < 220 > < 221 > PEPTIDE < 222 > (5) < 223 > Asp (OAIIyl) < 220 > < 221 > PEPTIDE < 222 > (5) < 223 > "This C-terminal amino acid is in the form of an ester with benzyl alcohol" < 400 > 39 Lys Glu Arg Val Asp 1 5 < 210 > 40 < 211 > 5 < 212 > PRT h 0 < 223 > Artificial Sequence < 220 > < 221 > PEPTIDE < 222 > (1) 15 < 223 > FMOC-Lys I < 220 > < 221 > PEPTIDE < 222 > (2) 20 < 223 > Glu (OtBu) < 220 > < 221 > PEPTIDO < 222 > (3) < 223 > Arg (PMC) < 220 > < 221 > PEPTIDE < 222 > (5) < 223 > "This C-terminal amino acid is in the form of an ester with benzyl alcohol" < 400 > 40 Lys Glu Arg Val Asp 1 5 < 210 > 41 < 211 > 5 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > PEPTIDE < 222 > (1) < 223 > FMOC-Lys < 220 > < 221 > PEPTIDE < 222 > (1 ) . . (5) < 223 > "Synthetic Peptides: The side chains of Lys in position 1 and Asp in position 5 are linked by an amide bond." < 220 > < 221 > PEPTIDE < 222 > (2) < 223 > Glu (OtBu) < 220 > < 221 > PEPTIDE < 222 > (3) < 223 > Arg (PMC) < 220 > < 221 > PEPTIDE < 222 > (1) < 223 > "Synthetic Peptides: The side chains of Lys in position 1 and Asp in position 5 are linked by an amide bond." < 220 > < 221 > PEPTIDO < 222 > (5) < 223 > "This C-terminal amino acid is in the form of an ester with benzyl alcohol" < 400 > 41 Lys Glu Arg Val Asp 1 S < 210 > 42 < 211 > 5 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > PEPTIDE < 222 > (1) < 223 > FMOC-Lys < 220 > < 221 > PEPTIDE < 222 > (1) .. (5) < 223 > "Synthetic peptide: The side chains of Lys in position 1 and Asp in position 5 are linked by an amide bond " < 221 > PEPTIDE < 222 > (2) < 223 > Glu (OtBu) < 220 > < 221 > PEPTIDE < 222 > (3) < 223 > Arg (PMC) < 220 > < 221 > PEPTIDE < 222 > (5) < 223 > "Synthetic peptide: The side chains of Lys in position 1 and Asp in position 5 are linked by an amide bond" < 400 > 42 Lys Glu Arg Val Asp 1, 5 < 210 > 43 < 211 > 5 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > PEPTIDO < 222 > (1) < 223 > FMOC-Lys (Alloc) < 220 > < 221 > PEPTIDE < 222 > (2) < 223 > Glu (OtBu) < 220 > < 221 > PEPTIDE < 222 > (3) < 223 > Arg (PMC) < 220 > < 221 > PEPTIDE < 222 > (5) < 223 > Asp (OAIIyl) < 220 > < 221 > PEPTIDE < 222 > (5) < 223 > "The C-terminal Asp is bound to chlorotryril resin" < 400 > 43 Lys Glu Arg Val Asp 1 5 < 210 > 44 < 211 > 5 < 212 > PRT < 223 > Artificial Sequence < 220 > < 221 > PEPTIDE < 222 > (1) < 223 > FMOC-Lys < 220 > < 221 > PEPTIDE < 222 > (2) < 223 > Glu (OtBu) < 220 > < 221 > PEPTIDE < 222 > (3) < 223 > Arg (PMC) < 220 > < 221 > PEPTIDE < 222 > (5) < 223 > "The C-terminal Asp is bound to chlorotryril resin." < 400 > 44 Lys Glu Arg Val Asp 1 5 < 210 > 45 < 211 > 5 < 212 > PRT < 213 > Artificial Sequence < 220 > < 221 > PEPTIDE < 222 > (1) < 223 > FMOC-Lys < 220 > < 221 > PEPTIDE < 222 > (1) < 223 > "The C-terminal Asp is bound to chlorotryril resin." < 220 > f < 221 > PEPTIDE 5 < 222 > (2) < 223 > Glu (OtBu) < 220 > < 221 > PEPTIDE < 223 > Arg (PMC) < 220 > < 221 > PEPTIDO 15 < 222 > (5) < 223 > "The C-terminal Asp is bound to chlorotryril resin." < 220 > < 221 > PEPTIDE 20 < 222 > (1) .. (5) < 223 > The side chains of Lys in position 1 and Asp in position 5 are linked by an amide bond < 400 > 45 Lys Glu Arg Val Asp 1 5 | < 210 > 46 5 < 211 > 34 < 212 > PRT < 213 > Artificial Sequence < 220 > ? < 223 > Artificial sequence description: HPTH analog < 220 > < 221 > PEPTIDE < 222 > (1) 15 < 223 > Product = Wing I < 220 > < 221 > MOD_RES < 222 > (8) 20 < 223 > Xaa = Norleucine (NIe) < 220 > < 221 > PEPTIDO < 222 > (18) < 223 > Product = Lys < 220 > < 221 > PEPTIDE < 222 > (18) < 223 > The side chain of Lys is linked by an amide bridge to the side chain of Asp < 220 > through an amide bridge to the < 223 > Product = Lys < 220 > f < 221 > PEPTIDE 5 < 222 > (26) < 223 > The Lys side chain is linked by an amide bridge to the side chain of Asp. < 220 > flO < 221 > PEPTIDE < 222 > (27) < 223 > Product = Leu < 220 > 15 < 221 > PEPTIDE < 222 > (30) f < 223 > Product = Asp < 220 > 20 < 221 > PEPTIDE < 222 > (30) < 223 > The side chain of Asp is linked by an amide bridge to the side chain of Lys < 400 > 46 Wing Val Ser Glu He Gln Leu Xaa His Asn Leu Gly Lys His Leu Asn 1 5 10 15 Ser Lys Glu Arg Val Asp Trp Leu Arg Lys Leu Leu Gln Asp Val His 20 25 30

Claims

NOVELTY OF THE INVENTION CLAIMS

1. - A method to prepare: wherein J, L, and M are fragments of linear peptide, Ki is absent or is a fragment of cyclic peptide, and K2 is a fragment of cyclic peptide; this method comprises the steps of: (1) preparing M - (R) by successively attaching amino acid residues suitably protected to a resin to provide: M- (R) III wherein (R) is a suitable peptide synthesis resin and M is a polypeptide fragment; (2) prepare separately by conventional peptide synthesis the protected N-terminal cyclic polypeptide fragment of formula IV PHN ^ / CC ^ H K2 IV where P is a suitable amine protecting group, (3) coupling with IV for provide a peptide of formula V (4) when the cyclic peptide K is absent, then (a) prepare the polypeptide fragments J and L as a single polypeptide of formula VI PHN. .L. 5 and coupling polypeptide VI to peptide V to provide a peptide of formula VII PHN. , / - S) Vil or, optionally, (b) add the protected individual amino acid elements of the J and L polypeptides consecutively to the peptide fragment of formula V, or, optionally, (c) ) preparing separately fragments of either J and L or both as polypeptide fragments and coupling these fragments to the growing peptide starting with the fragment of formula V; (5) when the cyclic peptide Ki is present, then (a) prepare separately the protected N-terminal cyclic polypeptide fragment 15 of formula VIII VIII wherein P is a suitable amine protecting group, by conventional peptide synthesis methods; (b) preparing a peptide fragment of formula IX PHNV ^ C02H IX and coupling the peptide fragment IX with the peptide V fragment to provide a peptide fragment of formula X PHN-LN N3) X; and (c) coupling the cyclic peptide fragment VIII with the peptide fragment of formula X to provide a peptide fragment of formula XI (d) preparing the peptide fragment of formula XII • 10 PHN, ^. C02H XII and coupling fragment XII with fragment XI; and (6) segment the resin and deprotect.

2. The method according to claim 1, Further characterized in that the peptide of formula I is a cyclic peptide compound of the formula | X-A1o-A ??- A12-A13-A14-A15-A16-A17-A ?8-A19-A20-A21-A22-A23-A24-A25-A25-A26-A27-Y wherein X is selected from the group consisting of (a) R a-A0-A A2-A3-A-A5-A6-A7-A8-A9-, (b) R1a-A2-A3-A4-A5-A6-A7-A8-Ag -, (c) R1 -A3-A4-A5-A6-A7-A8- 20 Ag-, (d) R1a-A4-A5-A6-A7-Aβ-Ag-, (f) R, a- Aß- A -Ag-Asp, (g) Rα-A7-Aβ-Ag-, (h) R1a-Aβ-Ag-, (i) Ria-Ag-, and (j) Ría-; Y is selected from the group consisting of (a) -R3, (b) -A28-R3, (c) -A28-A29-R3, (d) -A28-A29-A30-R3, (e) -A28- A2 -A3o-A3i-R3, (f) -A28- A29-A30-A31-A32-R3, (g) -A28-A29-A30-A31-A32-A33-R3, and (h) -A28-A2g -A3o-A31-A32- A33-A34-R3; RIA is H, alkyl, aralkyl or -COR2; R- is R? A or a group of formula R2 is alkyl, alkenyl, alkynyl, aryl, or aralkyl; R3 is a group of formula A35-OR4 or A35-NR4R5; R4 and R5 are independently H or lower alkyl; Re and Rg are independently H or alkyl; R7 is alkyl; Rs is H, alkyl or COR2; R10 is H or halogen; Rn is alkyl or aralkyl; m is 1, 2 or 3; n is 3 or h? 4; Ao is absent or is a peptide of one to six amino acid residues; Ai is Ser, Ala, Gly or D-Pro, or an equivalent amino acid; A is Ala, Val or Gly, or an equivalent amino acid; A3 is Ala, Ser, Gly or D-Pro, or an equivalent amino acid; A4 is Glu, Ala or Gly, or an equivalent amino acid; A lie, His, Ala or Gly, or an equivalent amino acid thereof; A6 is Ala, Gln, Gly or 15 D-Pro, or an equivalent amino acid thereof; A7 is Ala, Leu, Gly or an equivalent amino acid thereof; Aß is Leu, NIe, Gly or D-Pro, or a I equivalent amino acid thereof; Ag is His, Ala, D-Pro or Gly, or an equivalent amino acid thereof; A10 is Ala, Asn, Asp, Cys, homo-Cys, Glu, Gly, Lys, Orn, Ser, Thr, D-Pro, -NHCH (CH2) mNH2) CO- or -20 NHCH [(CH2) nC02H] CO -; An is Ala, Gly, Leu or Lys, or an equivalent amino acid thereof; A 2 is Ala or Gly, or an equivalent amino acid thereof; A13 is Ala, Asn, Asp, Cys, homo-Cys, Glu, Gly, Lys, Orn, Ser, Thr, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO-; A is Ala, Asn, Asp, Cys, homo-Cys, Glu, Gly, His, Lys, Orn, Ser, Thr, D-Pro, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO-; A15 is Ala, Gly, He, D-Pro or Leu, or an equivalent amino acid thereof; Aiß is Asn, Ala, Gly, D-Pro or Gln, or an amino acid ^ equivalent thereof; A17 is Ala, Asn, Asp, Cys, homo-Cys, Glu, Gly, 5 Lys, Orn, Ser, Thr, D-Pro, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO -; Ais is Asp, Cys, homo-Cys, Glu, His, Leu, Lys, Orn, NIe, Ser, Thr, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) "C02H] CO-; A19 is Arg or Glu, or an equivalent amino acid thereof; A2o is Arg or an equivalent amino acid thereof; A2? is Arg, Asp, Cys, homo-Cys, Glu, Lys Orn, Ser, h? Thr, Val, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO-; A22 is Asp, Cys, homo-Cys, Glu, His, Lys, Orn, Phe, Ser, Thr, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO-; A23 is Leu, Phe or Trp, or an equivalent amino acid thereof; A2 is Leu or an equivalent amino acid thereof; A25 is Arg, Asp, Cys, homo-Cys, Glu, His, Lys, Orn, D-Pro, Ser, Thr, -15 NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO-; A26 is Asp, Cys, homo- Cys, Glu, His, Lys, Orn, Ser, Thr, -NHCH (CH2) mNH2) CO- or - | NHCH [(CH2) nC02H] CO-; A27 is Leu or Lys, or an equivalent amino acid thereof; A28 is He or Leu; or an equivalent amino acid thereof; A2g is Ala, Asp, Cys, homo-Cys, Glu, Gln, Lys, Orn, Ser, Thr, -NHCH (CH2) mNH2) CO-20 or -NHCH [(CH2) nC02H] CO-; A30 is Asp, Cys, homo-Cys, Glu, Gly, Lys, Orn, Ser, Thr, -NHCH (CH2) mNH2) CO- or -NHCH [(CH2) nC02H] CO-; A31 is Lie, Leu or Val, or an equivalent amino acid thereof; A32 is His, or an equivalent amino acid thereof; A 3 is Asn or Thr, or an amino acid equivalent of same; and A34 is Ala or Phe, or an equivalent amino acid thereof; A35 is absent or is a peptide of 1 to 4 amino acids; and the side chains of at least one of the following pairs of amino acid residues, A 0 and AM, A13 and A17, A14 and A18, A17 and A21, A18 and A22, A21 and A25, A25 and A2g and A26 and A30 are 5 linked by an amide, ester, disulfide or lanthionine bond to form a bridge, and the side chain of each of the following amino acid residues, A10, A? 3, Au, A17, A? 8, A2 ?, A22 , A25, A2ß, A2g, and A 0 contribute, to the most, to the formation of a single bridge; provided that when the side chains of the following pairs of amino acid residues, A13 and h? A17 or A2ß and A30 are linked by an amide, disulfide or lanthionine bond to form a bridge, then the side chains of at least one of the following pairs of amino acid residues, A10 and Au, A14 and Ais, A17 and A2? , Aiß and A22, A21 and A25 and A25 and A2g are also linked by an amide, ester, disulfide or lanthionine linkage.

3. The method according to claim 1 for preparing cycle (Lys18-Asp22) [Ala1, NIe8, Lys18, Asp22, Leu27] hPTH (1-31) NH2 whose The method comprises the steps of (a) preparing the peptide fragment bound to resin A: Fmoc-Trp resin (Boc) -Leu-Arg (PMC) -Lys (Boc) -Leu-Leu-Gln (Trt) - Asp (OtBu) -Val-NH (A); (b) preparing the cyclic peptide fragment Fmoc-Lys-Glu (OtBu) -Gar (Pmc) -Val-Asp-OH (B); (c) coupling the peptide A fragment from step (a) with the peptide B fragment from step (b) to form the intermediate peptide AB Resin of Fmoc-Lys-Glu-Arg-Val-Asp-Trp (Boc) -Leu-Arg (Pmc) - Lys (Boc) -Leu-Leu-Gln (Trt) -Asp (OtBu) -Val-NH (d) sequentially add peptide AB Ser (OtBu), Asn (Trt), Leu, His (Trt) and Lys (Boc) to form the intermediate peptide C: Resin of Fmoc-Lys (Boc) -His (Trt) -Leu-Asn (Trt) -Ser (OtBu) -Lys-Glu-Arg-Val-Asp-Tf (Boc) -Leu-Arg (PMC) -Lys ( Boc) -Leu-Leu-Gln (Trt) -Asp (OtBu) -Val-NH-resin (C); (e) prepare the peptide fragment D: Fmoc-Ala-Val-Ser (OtBu) -Glu (OtBu) -lle-Gln (Trt) -Leu-Nle-His (Trt) -Asn (Trt) -Leu-Gly -OH D); (f) coupling the C and D fragments to form the peptide attached to resin E: Resin of Fmoc-Ala-Val-Ser (? TBu) -Glu (? TBu) -lle-Gln (Trt) -Leu-Nle-His (Trt) -Asn (Trt) -Leu-Gly-Lys (Boc) - His (Trt) -Leu-Asn (Trt) -Ser (? TBu) -Lys-Glu-Arg-Val-Asp-T (Boc) -Leu-Arg (PMC) -Lys (Boc) -Leu-Leu-Gln (Trt) -Asp (? TBu) -Val-NH (E); and (g) segment the resin and deprotect.

4. A suitable peptide as an intermediate for carrying out the method of claim 3, selected from the group consisting of: resin of Fmoc-Trp (OtBu) -Leu-Arg (Pmc) -Lys- (Boc) -Leu-Leu-Gln (Trt) -Asp (OtBu) -Val- NH, Cyclo-resin (Lys18-Asp22) Fmoc-Lys -Glu (OtBu) -Arg (Pmc) -Val-Asp- Trp (Boc) -Leu-Arg (Pmc) -Lys (Boc) -Leu-Leu-Gln (Trt) -Asp (OtBu) -Val-NH, resin ) cycle (Lys18-Asp22) Fmoc-Lys- (Boc) -His (Trt) -Leu-Asn (Trt) -Ser (OtBu) -Lys- 5 Glu (OtBu) -Arg (Pmc) -Val-Asp- Trp (Boc) -Leu-Arg (Pmc) -Lys (Boc) -Leu-Leu-Gln (Trt) -Asp (OtBu) -Val-NH, Fmoc-Ala-Val-Ser (OtBu) -Glu (OtBu) -lle-Gln (Trt) - Leu-Nle-His (Trt) -Asn (Trt) -Leu-Gly-OH, cycle resin (Lys18-Asp22) Fmoc-Ala-Val-Ser (OtBu) -Glu (OtBu) ) -lle-Gln (Trt) -Leu-Nle-His (Trt) -Asn (Trt) -Leu-Gly-Lys (Boc) -His (Trt) -Leu-Asn (Trt) -Ser (OtBu) -Lys -Glu (OtBu) -Arg (Pmc) -Val-Asp-h? Trp (Boc) -Leu-Arg (Pmc) -Lys (Boc) -Leu-Leu-Gln (Trt) -Asp (OtBu) -Val-NH, Boc-Val-Asp (OAIIyl) -OBzl, HCI.Val- Asp (OAIIyl) -OBzl, Fmoc-Arg (Pmc) -Val-Asp (OAIIyl) -OBzl, Arg (Pmc) -Val-Asp (OAIIyl) -OBzl, Fmoc-Glu (OtBu) -Arg (Pmc) - Val -Asp (OAIIyl) -OBzl, Glu (OtBu) -Arg (Pmc) -Val-Asp (OAIIyl) -OBzl, Fmoc- Lys (Alloc) -Glu (OtBu) -Arg (Pmc) -Val-Asp (OAIIyl) -OBzl, Fmoc-Lys-Glu (OtBu) -15 Arg (Pmc) -Val-Asp-OBzl, Fmoc-cycle (Lys-Asp) Lys-Glu (OtBu) -Arg (Pmc) -Val- Asp-OBzl, Fmoc-cycle (Lys-Asp) Lys-Glu (OtBu) -Arg (Pmc) -Val-Asp-OH, resin of Fmoc-Lys (Alloc) -Glu (OtBu) -Arg (Pmc) -Val-Asp (OAIIyl ) -chlorotritil, Fmoc-Lys-Glu resin (OtBu) -Arg (Pmc) -Val-Asp-chlorotritil, and Fmoc-cyclo (Lys-Asp) resin Lys-Glu (OtBu) -Arg (Pmc) -Val -Asp-chlorotritil.