Method for preparation of PSMA conjugates

Abstract

The present disclosure relates to methods for preparing compounds useful in the treatment of diseases such as cancer in mammals. In particular, the invention described herein relates to methods for preparing compounds that can target PSMA-expressing cells and are useful in the treatment of diseases caused by PSMA-expressing cells, such as prostate cancer.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of and priority to U.S. Provisional Patent Application No. 62 / 850,119, filed May 20, 2019, U.S. Provisional Patent Application No. 62 / 910,777, filed October 4, 2019, and U.S. Provisional Patent Application No. 62 / 912,353, filed October 8, 2019, the entire disclosures of which are incorporated herein by reference.

[0002] The present disclosure relates to methods for preparing compounds useful for treating diseases such as cancer in mammals. In particular, the invention described herein relates to methods for preparing compounds that can target PSMA-expressing cells and are useful for treating diseases caused by PSMA-expressing cells, such as prostate cancer. [Background technology]

[0003] The prostate is a male reproductive organ that functions to produce and store semen, which provides nutrients and fluids for the survival of spermatozoa introduced into the vagina during reproduction. Like other tissues, the prostate can develop either malignant (cancerous) or benign (non-cancerous) tumors. In fact, prostate cancer is one of the most common male cancers in Western society and the second most common form of malignancy among American men. Current methods of treating prostate cancer include hormone therapy, radiation therapy, surgery, chemotherapy, photodynamic therapy, and combination therapy. However, many of these treatments affect the quality of life of male patients diagnosed with prostate cancer, especially those over the age of 50. For example, the use of hormonal drugs is often accompanied by side effects such as osteoporosis and liver damage. Such side effects can be mitigated by the use of treatments that are more selective or specific to the tissues causing the condition and that avoid non-target tissues such as bone or liver.

[0004] Prostate-specific membrane antigen (PSMA) is a biomarker that is overexpressed in prostate cancer. PSMA is overexpressed in malignant prostate tissues compared to other organs of the human body, such as the kidney, proximal small intestine, and salivary glands. PSMA is also expressed in neovasculature in many non-prostatic solid tumors, including lung, colon, breast, renal, hepatic, and pancreatic cancers, but not in normal vasculature. PSMA is also minimally expressed in the brain. PSMA is a type II cell surface membrane-associated glycoprotein with a molecular weight of approximately 110 kD, containing an intracellular segment (amino acids 1-18), a transmembrane domain (amino acids 19-43), and an extensive extracellular domain (amino acids 44-750). The function of the intracellular segment and the transmembrane domain is currently thought to be unimportant, while the extracellular domain is involved in several different activities. For example, PSMA plays a role in the central nervous system, where it metabolizes N-acetyl-aspartylglutamate (NAAG) to glutamate and N-acetylaspartate. PSMA also plays a role in the proximal small intestine, where it removes gamma-linked glutamate from poly-gamma-glutamate folates and alpha-linked glutamate from peptides and small molecules. However, the specific function of PSMA on prostate cancer cells remains unresolved.

[0005] Unlike many other membrane-bound proteins, PSMA is rapidly taken up into cells in a manner similar to cell surface-bound receptors such as vitamin receptors.PSMA can be internalized through clathrin-coated pits and then recycled to the cell surface or transported to lysosomes.Therefore, diagnostic, imaging, and therapeutic drugs can be targeted to PSMA for delivery into PSMA-expressing cells, such as prostate cancer cells.

[0006] Described herein are methods for preparing compounds capable of binding to PSMA. Also described herein are methods for preparing compounds capable of targeting PSMA for diagnosis, imaging, and delivery of therapeutic agents.

[0007] It has been unexpectedly discovered that the conjugates prepared by the methods described herein exhibit high affinity for PSMA. It has also been discovered that the compounds prepared by the methods described herein are effective in treating diseases caused by pathogenic cells that express PSMA, such as prostate cancer cells.

[0008] It is disclosed that certain conjugates show high affinity to PSMA. It is also disclosed that the conjugates are effective in treating diseases caused by pathogenic cells expressing PSMA, such as prostate cancer cells. One such conjugate is described in WO 2015 / 055318 A1: [ka] (also known as PSMA-617).

[0009] Due to the demand for pharmaceuticals such as PSMA-617, there is a need to provide synthetic methods that can supply large quantities of such products at lower cost and with high purity. Described herein are methods for preparing compounds capable of binding to PSMA, such as PSMA-617. Summary of the Invention

[0010] In one aspect, the disclosure provides methods for preparing compounds useful in the treatment of disease, particularly cancer, in mammals, including humans.

[0011] In one embodiment, the disclosure provides methods for preparing compounds for treating abnormal cell growth in a mammal, including a human, in need of such treatment.

[0012] In another embodiment, the abnormal cell growth is cancer, hi another embodiment, the cancer is prostate cancer, metastatic prostate cancer, and metastatic castration-resistant prostate cancer.

[0013] In another aspect, the present disclosure provides a compound of formula I [ka] A method for preparing the compound is provided.

[0014] Further embodiments, features, and advantages of the present disclosure will become apparent from the following detailed description and through the practice of the present disclosure. The compounds of the present disclosure may be described as embodiments in any of the following enumerated clauses. It will be understood that any of the embodiments described herein can be used in conjunction with any other embodiment described herein to the extent that the embodiments are not mutually exclusive.

[0015] 1. Formula I [ka] A process for preparing a compound of the formula: Formula A [ka] contacting the resin-based compound of formula A' with a first deprotecting agent in a polar aprotic solvent to produce a compound of formula A' [ka] or providing a resin-based compound of b. A resin-based compound of formula A' is mixed with a resin-based compound of formula B [ka] in the presence of an organic solvent and a base to form a compound of formula C [ka] or providing a resin-based compound of c. contacting the resin-based compound of formula C with a second deprotecting agent in the presence of an organic solvent to produce a compound of formula C' [ka] or providing a resin-based compound of d. The resin-based compound of formula C' is reacted with PG in the presence of a polar aprotic solvent, a coupling agent and a base. 1 -3-(2-naphthyl)-L-alanine to form a compound of formula D [ka] or providing a resin-based compound of e. contacting the resin-based compound of formula D with a first deprotecting agent in a polar aprotic solvent to produce a compound of formula D' [ka] or providing a resin-based compound of f. The resin-based compound of formula D' is reacted with PG in the presence of a polar aprotic solvent, a coupling agent and a base. 1 - contacting with tranexamic acid to form a compound of formula E [ka] or providing a resin-based compound of g. contacting the resin-based compound of formula E with a first deprotecting agent in a polar aprotic solvent to produce a compound of formula E' [ka] or providing a resin-based compound of h. Resin-based compound of formula E' with a compound of formula E 2’ [ka] in the presence of a polar aprotic solvent, a coupling agent, and a base to obtain a compound of formula F [ka] or providing a resin-based compound of i. contacting a resin-based compound of formula F with a third deprotecting and / or cleaving agent in an organic solvent; In the ceremony, P.G. 1 and P.G. 2 is an amine protecting group, LG is a leaving group, and R 1 is C1-C4 alkyl or cyclohexyl.

[0016] 2. The method of claim 1, wherein the first deprotecting agent in step (a) is selected from the group consisting of piperidine, morpholine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), diethylamine (DEA), dicyclohexamine, 4-methylpiperidine (4MP), tris(2-aminoethyl)amine, pyridine and collidine.

[0017] 3. The method according to any one of the preceding claims, wherein the first deprotecting agent in step (e) is selected from the group consisting of piperidine, morpholine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), diethylamine (DEA), dicyclohexamine, 4-methylpiperidine (4MP), tris(2-aminoethyl)amine, pyridine and collidine.

[0018] 4. The method according to any one of the preceding claims, wherein the first deprotecting agent in step (g) is selected from the group consisting of piperidine, morpholine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), diethylamine (DEA), dicyclohexamine, 4-methylpiperidine (4MP), tris(2-aminoethyl)amine, pyridine and collidine.

[0019] 5. The method of any one of the preceding claims, wherein the first deprotecting agent in step (a) is piperidine.

[0020] 6. The method of any one of the preceding claims, wherein the first deprotecting agent in step (e) is piperidine.

[0021] 7. The method of any one of the preceding claims, wherein the first deprotecting agent in step (g) is piperidine.

[0022] 8. The method according to any one of the preceding claims, wherein the polar aprotic solvent in step (a) is selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO).

[0023] 9. The method according to any one of the preceding claims, wherein the polar aprotic solvent in step (d) is selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO).

[0024] 10. The method according to any one of the preceding claims, wherein the polar aprotic solvent in step (e) is selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO).

[0025] 11. The method according to any one of the preceding claims, wherein the polar aprotic solvent in step (f) is selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO).

[0026] 12. The method according to any one of the preceding claims, wherein the polar aprotic solvent in step (g) is selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO).

[0027] 13. The method according to any one of the preceding claims, wherein the polar aprotic solvent in step (h) is selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO).

[0028] 14. The method according to any one of the preceding claims, wherein the polar aprotic solvent in step (a) is dimethylformamide (DMF).

[0029] 15. The method according to any one of the preceding claims, wherein the polar aprotic solvent in step (d) is dimethylformamide (DMF).

[0030] 16. The method according to any one of the preceding claims, wherein the polar aprotic solvent in step (e) is dimethylformamide (DMF).

[0031] 17. The method according to any one of the preceding claims, wherein the polar aprotic solvent in step (f) is dimethylformamide (DMF).

[0032] 18. The method according to any one of the preceding claims, wherein the polar aprotic solvent in step (g) is dimethylformamide (DMF).

[0033] 19. The method according to any one of the preceding claims, wherein the polar aprotic solvent in step (h) is dimethylformamide (DMF).

[0034] 20. The base in step (b) is N,N-diisopropylethylamine ( i 2. The method of any one of the preceding claims, wherein the cation exchanger is selected from the group consisting of PrNEt), triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine.

[0035] 21. The base in step (d) is N,N-diisopropylethylamine ( i 2. The method of any one of the preceding claims, wherein the cation exchanger is selected from the group consisting of PrNEt), triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine.

[0036] 22. The base in step (f) is N,N-diisopropylethylamine ( i 2. The method of any one of the preceding claims, wherein the cation exchanger is selected from the group consisting of PrNEt), triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine.

[0037] 23. The base in step (h) is N,N-diisopropylethylamine ( i 2. The method of any one of the preceding claims, wherein the cation exchanger is selected from the group consisting of PrNEt), triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine.

[0038] 24. The base in step (b) is N,N-diisopropylethylamine ( i 2. The method of any one of the preceding claims, wherein the cytoplasmic ion transporter is PrNEt.

[0039] 25. The base in step (d) is N,N-diisopropylethylamine ( i 2. The method of any one of the preceding claims, wherein the cytoplasmic ion transporter is PrNEt.

[0040] 26. The base in step (f) is N,N-diisopropylethylamine ( i 2. The method of any one of the preceding claims, wherein the cytoplasmic ion transporter is PrNEt.

[0041] 27. The base in step (h) is N,N-diisopropylethylamine ( i 2. The method of any one of the preceding claims, wherein the cytoplasmic ion transporter is PrNEt.

[0042] 28. The method of any one of the preceding claims, wherein the second deprotecting agent in step (c) is selected from the group consisting of trifluoroacetic acid (TFA), acetic acid, trifluoroethanol, hexafluoroisopropanol, dichloromethane, and combinations thereof.

[0043] 29. The method of any one of the preceding claims, wherein the second deprotecting agent in step (c) is trifluoroacetic acid (TFA) or a mixture of acetic acid and trifluoroethanol.

[0044] 30. The coupling agent in step (d) is benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT), N-[(5-chloro-3-oxide-1H-benzotriazol-1-yl)-4-morpholinylmethylene]-N-methylmethanaminium hexafluorophosphate (HBTU), 4,5-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM), 1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (HDMC), 1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (COMU), dimethylamino(triazolo[4,5-b]pyridin-3-yloxy)methylidene]-dimethylazanium; tetrafluoroborate (TATU), N,N,N',N'-tetramethyl-S-(1-oxido-2-pyridyl)thiouronium tetrafluoroborate (TOTT), N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), 1-propanephosphonic anhydride (T3P), and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM).

[0045] 31. The coupling agent in step (f) is benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT), N-[(5-chloro-3-oxide-1H-benzotriazol-1-yl)-4-morpholinylmethylene]-N-methylmethanaminium hexafluorophosphate (HBTU), 4,5-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM), 1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (HDMC), 1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (COMU), dimethylamino(triazolo[4,5-b]pyridin-3-yloxy)methylidene]-dimethylazanium; tetrafluoroborate (TATU), N,N,N',N'-tetramethyl-S-(1-oxido-2-pyridyl)thiouronium tetrafluoroborate (TOTT), N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), 1-propanephosphonic anhydride (T3P), and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM).

[0046] 32. The coupling agent in step (h) is benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT), N-[(5-chloro-3-oxide-1H-benzotriazol-1-yl)-4-morpholinylmethylene]-N-methylmethanaminium hexafluorophosphate (HBTU), 4,5-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM), 1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (HDMC), 1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (COMU), dimethylamino(triazolo[4,5-b]pyridin-3-yloxy)methylidene]-dimethylazanium; tetrafluoroborate (TATU), N,N,N',N'-tetramethyl-S-(1-oxido-2-pyridyl)thiouronium tetrafluoroborate (TOTT), N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), 1-propanephosphonic anhydride (T3P), and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM).

[0047] 33. The method according to any one of the preceding claims, wherein the coupling agent in step (d) is benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP).

[0048] 34. The method according to any one of the preceding claims, wherein the coupling agent in step (f) is benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP).

[0049] 35. The method according to any one of the preceding claims, wherein the coupling agent in step (h) is benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP).

[0050] 36. The method according to any one of the preceding claims, wherein the third deprotecting agent in step (i) is selected from the group consisting of trifluoroacetic acid (TFA), hydrofluoric acid (HF), triflic acid (TfOH), a trifluoroacetic acid (TFA) / water / triisopropylsilane mixture, a trifluoroacetic acid (TFA) / dichloromethane (DCM) mixture, triethylsaline (TES), indole, a phenol / anisole mixture, and thioanisole.

[0051] 37. The method according to any one of the preceding claims, wherein the third deprotecting agent in step (i) is a trifluoroacetic acid (TFA) / water / triisopropylsilane mixture.

[0052] 38. The method according to any one of the preceding claims, wherein the cleavage reagent in step (i) is trifluoroacetic acid (TFA) or a trifluoroacetic acid (TFA) / water / triisopropylsilane mixture.

[0053] 39. The method according to any one of the preceding claims, wherein the cleavage reagent in step (i) is a trifluoroacetic acid (TFA) / water / triisopropylsilane mixture.

[0054] 40. The method according to any one of the preceding claims, wherein the organic solvent in step (b) is selected from the group consisting of CH2Cl2, THF, diethyl ether, MeOH, EtOH, acetone, and acetonitrile.

[0055] 41. The method according to any one of the preceding claims, wherein the organic solvent in step (c) is selected from the group consisting of CH2Cl2, THF, diethyl ether, MeOH, EtOH, acetone, and acetonitrile.

[0056] 42. The method according to any one of the preceding claims, wherein the organic solvent in step (i) is selected from the group consisting of CH2Cl2, THF, diethyl ether, MeOH, EtOH, acetone, and acetonitrile.

[0057] 43. The method according to any one of the preceding claims, wherein the organic solvent in step (b) is CH2Cl2.

[0058] 44. The method according to any one of the preceding claims, wherein the organic solvent in step (c) is CH2Cl2.

[0059] 45. The method according to any one of the preceding claims, wherein the organic solvent in step (i) is CH2Cl2.

[0060] 46. ​​PG 1 Item 11. The method according to any one of the preceding claims, wherein is 9-fluorenylmethyl-carbonyl (Fmoc) or t-butylcarbonyl (Boc).

[0061] 47. PG 1 The method according to any one of the preceding claims, wherein is 9-fluorenylmethyl-carbonyl (Fmoc).

[0062] 48. PG 2 Item 11. The method of any one of the preceding, wherein is monomethoxytrityl (MMt) or 4-methyltrityl (Mtt).

[0063] 49. PG 2 Item 11. The method of any one of the preceding, wherein is 4-methyltrityl (Mtt).

[0064] 50. LG, [ka] and * Item 11. The method of any one of the preceding, wherein represents a point of attachment to the remainder of the compound.

[0065] 51. R 1Item 11. The method of any one of the preceding, wherein is t-butyl.

[0066] 52. Formula 1 [ka] A method for preparing a compound of the formula: a. Formula I [ka] contacting the resin-based compound of formula 2 with a first deprotecting agent in the presence of a solvent to form a compound of formula 2 [ka] or providing a resin-based compound of b. The resin-based compound of formula 2 is mixed with a compound of formula 3 [ka] in the presence of an organic solvent and a base to form a compound of formula 4 [ka] or providing a resin-based compound of c. contacting the resin-based compound of formula 4 with a second deprotecting agent in the presence of an organic solvent to produce a compound of formula 5 [ka] or providing a resin-based compound of d. The resin-based compound of formula 5 is reacted with a compound of formula 6 [ka] in the presence of a solvent, a coupling agent, and optionally a base and / or an additive to obtain a compound of formula 7 [ka] or providing a resin-based compound of e. contacting the resin-based compound of formula 7 with a third deprotecting agent in the presence of a solvent to produce a compound of formula 8. [ka] or providing a resin-based compound of f. The resin-based compound of formula 8 is reacted with a compound of formula 9 [ka] in the presence of a solvent, a coupling agent, and optionally a base and / or an additive to obtain a compound of formula 10 [ka] or providing a resin-based compound of g. contacting the resin-based compound of formula 10 with at least one third deprotecting and / or cleaving agent in the presence of an organic solvent to provide a compound of formula I. one or more of In the formula, PG, PG 1 and P.G. 4 is an amine protecting group, PG 2 , P.G. 3 , and P.G. 5 is a carbonyl protecting group, and LG of compound C is a leaving group.

[0067] 53. P.G., P.G. 1 and P.G. 4 53. The method of claim 52, wherein is selected from the group consisting of 9-fluorenylmethyl-carbonyl (Fmoc), benzyl carbamate (Cbz), t-butyloxycarbonyl (Boc), acetamide, trifluoroacetamide, p-toluenesulfonamide, triphemylmethyl, monomethoxytrityl (MMt), and 4-methyltrityl (Mtt).

[0068] 54. The method of claim 52 or 53, wherein PG is 9-fluorenylmethyl-carbonyl.

[0069] 55. PG 1 55. The method of any one of clauses 52 to 54, wherein is monomethoxytrityl (MMt) or 4-methyltrityl (Mtt).

[0070] 56. PG 1 56. The method of any one of clauses 52 to 55, wherein is 4-methyltrityl (Mtt).

[0071] 57. PG 4 57. The method according to any one of clauses 52 to 56, wherein is 9-fluorenylmethyl-carbonyl.

[0072] 58. PG 2 , P.G. 3 , and P.G. 5 58. The method of any one of clauses 52 to 57, wherein each is t-butyl.

[0073] 59. The method according to any one of clauses 52 to 58, wherein LG of compound C is selected from imidazolyl, 4-nitrophenoxy, and phenoxy.

[0074] 60. The method of any one of clauses 52 to 59, wherein the first deprotecting agent of step (a) comprises a reagent or mixture of reagents capable of removing a protecting group selected from the group consisting of 9-fluorenylmethyl-carbonyl (Fmoc), benzyl carbamate (Cbz), t-butyloxycarbonyl (Boc), acetamide, trifluoroacetamide, p-toluenesulfonamide, triphemylmethyl, monomethoxytrityl (MMt), and 4-methyltrityl (Mtt).

[0075] 61. The method of any one of clauses 52 to 60, wherein the first deprotecting agent of step (a) comprises a reagent or mixture of reagents capable of removing 9-fluorenylmethyl-carbonyl.

[0076] 62. The method of any one of clauses 52 to 61, wherein the first deprotecting agent in step (a) is selected from the group consisting of piperidine, morpholine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), diethylamine (DEA), dicyclohexamine, 4-methylpiperidine (4MP), tris(2-aminoethyl)amine, pyridine, and collidine.

[0077] 63. The method of any one of clauses 52-62, wherein the first deprotecting agent in step (a) is piperidine.

[0078] 64. The method of any one of clauses 52 to 63, wherein the solvent in step (a) is a polar aprotic solvent.

[0079] 65. The method of any one of clauses 52 to 64, wherein the solvent in step (a) is selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO).

[0080] 66. The base in step (b) is N,N-diisopropylethylamine ( i 66. The method of any one of clauses 52 to 65, wherein the aryl group is selected from the group consisting of PrNEt), triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine.

[0081] 67. The method of any one of clauses 52-66, wherein the organic solvent in step (b) is selected from the group consisting of CH2Cl2, THF, diethyl ether, MeOH, EtOH, acetone, and acetonitrile.

[0082] 68. The method of any one of clauses 52 to 67, wherein the second deprotecting agent of step (c) comprises a reagent or mixture of reagents capable of removing a protecting group selected from the group consisting of 9-fluorenylmethyl-carbonyl (Fmoc), benzyl carbamate (Cbz), t-butyloxycarbonyl (Boc), acetamide, trifluoroacetamide, p-toluenesulfonamide, triphemylmethyl, monomethoxytrityl (MMt), and 4-methyltrityl (Mtt).

[0083] 69. The method of any one of clauses 52-68, wherein the second deprotecting agent of step (c) comprises a reagent or mixture of reagents capable of removing 4-methyltrityl (Mtt).

[0084] 70. The method of any one of clauses 52-69, wherein the second deprotecting agent in step (c) is selected from the group consisting of trifluoroacetic acid (TFA), acetic acid, 2,2,2-trifluoroethanol, hexafluoroisopropanol, and combinations thereof.

[0085] 71. The method of any one of clauses 52-70, wherein the second deprotecting agent in step (c) is a mixture of acetic acid and 2,2,2-trifluoroethanol.

[0086] 72. The method of any one of clauses 52-71, wherein the organic solvent in step (c) is selected from the group consisting of CH2Cl2, THF, diethyl ether, MeOH, EtOH, acetone, and acetonitrile.

[0087] 73. The coupling agent in step (d) is selected from the group consisting of benzotriazol-1-yloxy-tris(dimethylamino)-phosphonium hexafluorophosphate (BOP), benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), 7-aza-benzotriazol-1-yloxy-tripyrrolidino-phosphonium hexafluorophosphate (PyAOP), ethylcyano(hydroxyimino)acetato-O2)-tri-(1-pyrrolidinyl)-phosphonium hexafluorophosphate (PyO xim), bromo-tripyrrolidino-phosphonium hexafluorophosphate (PyBrOP), 3-(diethoxy-phosphoryloxy)-1,2,3-benzo[d]triazin-4(3H)-one (DEPBT), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), N-[(5-chloro-3-oxido-1H-benzotriazol-1-yl)-4-morpholinylmethylene]-N-methylmethanaminium hexafluorophosphate (HDMC), 1-cyano-2-ethoxyethyl 1H-benzotriazol-1-yl, 2-(6-chloro-1H-benzotriazol-1-yl)-N,N,N',N'-tetramethylaminium hexafluorophosphate (HBTU), 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT), N-[(5-chloro-3-oxido-1H-benzotriazol-1-yl)-4-morpholinylmethylene]-N-methylmethanaminium hexafluorophosphate (HDMC), 1-cyano-2-ethoxyethyl 1H-benzotriazol-1-yl, 2-(6-chloro-1H-benzotriazol-1-yl)-4-morpholinylmethylene]-N-methylmethanaminium hexafluorophosphate (HDMC), Si-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (COMU), dimethylamino(triazolo[4,5-b]pyridin-3-yloxy)methylidene]-dimethylazanium tetrafluoroborate (TATU), N,N,N',N'-tetramethyl-S-(1-oxido-2-pyridyl)thiouronium tetrafluoroborate (TOTT), tetramethylfluoroformamidinium hexafluorophosphate (TFFH), N-ethoxycarbonyl-2-ethoxy-1,73. The method according to any one of clauses 52 to 72, wherein the aryl group is selected from the group consisting of 2-dihydroquinoline (EEDQ), 1-propanephosphonic anhydride (T3P), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM), dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), (N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDAC), and 1,1'-carbonyldimidizole (CDI).

[0088] 74. The base in step (d) is N,N-diisopropylethylamine ( i 74. The method of any one of clauses 52 to 73, wherein the aryl group is selected from the group consisting of PrNEt), triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine.

[0089] 75. The method of any one of clauses 52-74, wherein the additive in step (d), if present, is selected from the group consisting of 1-hydroxybenzotriazole (HOBt), 1-hydroxybenzotriazole-6-sulfonamidomethyl resin HCl (HOBt-6-sulfonamidomethyl resin HCl), hydroxy-3,4-dihydro-4-oxo-1,2,3-benzotriazine (HOOBt), N-hydroxysuccinimide (NHS), 1-hydroxy-7-aza-1H-benzotriazole (HOAt), ethyl 2-cyano-2-(hydroximino)acetate, and 4-(N,N-dimethylamino)pyridine (DMAP).

[0090] 76. The method of any one of clauses 52 to 75, wherein the solvent in step (d) is a polar aprotic solvent.

[0091] 77. The method of any one of clauses 52 to 76, wherein the solvent in step (d) is selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), a dichloromethane / dimethylformamide mixture, acetonitrile (ACN), an acetonitrile / dimethylformamide mixture, and dimethylsulfoxide (DMSO).

[0092] 78. The method of any one of clauses 52 to 77, wherein the third deprotecting agent of step (e) comprises a reagent or mixture of reagents capable of removing a protecting group selected from the group consisting of 9-fluorenylmethyl-carbonyl (Fmoc), benzyl carbamate (Cbz), t-butyloxycarbonyl (Boc), acetamide, trifluoroacetamide, p-toluenesulfonamide, triphemylmethyl, monomethoxytrityl (MMt), and 4-methyltrityl (Mtt).

[0093] 79. The method of any one of clauses 52-78, wherein the third deprotecting agent of step (e) comprises a reagent or mixture of reagents capable of removing 9-fluorenylmethyl-carbonyl.

[0094] 80. The method of any one of clauses 52-79, wherein the first deprotecting agent in step (e) is selected from the group consisting of piperidine, morpholine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), diethylamine (DEA), dicyclohexamine, 4-methylpiperidine (4MP), tris(2-aminoethyl)amine, pyridine, and collidine.

[0095] 81. The method of any one of clauses 52-80, wherein the first deprotecting agent in step (e) is piperidine.

[0096] 82. The method of any one of clauses 52 to 81, wherein the solvent in step (e) is a polar aprotic solvent.

[0097] 83. The method of any one of clauses 52 to 82, wherein the solvent in step (e) is selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO).

[0098] 84. The coupling agent in step (f) is benzotriazol-1-yloxy-tris(dimethylamino)-phosphonium hexafluorophosphate (BOP), benzotriazol-1-yloxy-tripyrrolidinophosphonium hexafluorophosphate (PyBOP), 7-aza-benzotriazol-1-yloxy-tripyrrolidino-phosphonium hexafluorophosphate (PyAOP), ethylcyano(hydroxyimino)acetato-O2)-tri-(1-pyrrolidinyl)-phosphonium hexafluorophosphate (PyOx im), bromo-tripyrrolidino-phosphonium hexafluorophosphate (PyBrOP), 3-(diethoxy-phosphoryloxy)-1,2,3-benzo[d]triazin-4(3H)-one (DEPBT), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), 2-(1H-benzotriazole -1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), 2-(6-chloro-1H-benzotriazol-1-yl)-N,N,N',N'-tetramethylaminium hexafluorophosphate (HCTU), 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT), N-[(5-chloro-3-oxido-1H-benzotriazol-1-yl)-4-morpholinylmethylene]-N-methylmethanaminium hexafluorophosphate (HDMC), 1-cyano-2-ethoxy -2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (COMU), dimethylamino(triazolo[4,5-b]pyridin-3-yloxy)methylidene]-dimethylazanium tetrafluoroborate (TATU), N,N,N',N'-tetramethyl-S-(1-oxido-2-pyridyl)thiouronium tetrafluoroborate (TOTT), tetramethylfluoroformamidinium hexafluorophosphate (TFFH), N-ethoxycarbonyl-2-ethoxy-1,84. The method according to any one of clauses 52 to 83, wherein the aryl group is selected from the group consisting of 2-dihydroquinoline (EEDQ), 1-propanephosphonic anhydride (T3P), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM), dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), (N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDAC), and 1,1'-carbonyldimidizole (CDI).

[0099] 85. When the base in step (f) is present, it is N,N-diisopropylethylamine ( i 85. The method of any one of clauses 52 to 84, wherein the aryl group is selected from the group consisting of PrNEt), triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine.

[0100] 86. The method according to any one of items 52 to 85, wherein the additive in step (f), if present, is selected from the group consisting of 1-hydroxybenzotriazole (HOBt), 1-hydroxybenzotriazole-6-sulfonamidomethyl resin HCl (HOBt-6-sulfonamidomethyl resin HCl), hydroxy-3,4-dihydro-4-oxo-1,2,3-benzotriazine (HOOBt), N-hydroxysuccinimide (NHS), 1-hydroxy-7-aza-1H-benzotriazole (HOAt), ethyl 2-cyano-2-(hydroximino)acetate, and 4-(N,N-dimethylamino)pyridine (DMAP).

[0101] 87. The method of any one of clauses 52 to 86, wherein the solvent in step (f) is a polar aprotic solvent.

[0102] 88. The method of any one of clauses 52 to 87, wherein the solvent in step (f) is a polar aprotic solvent selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO).

[0103] 89. The method of any one of clauses 52 to 88, wherein the third deprotecting agent of step (g) is a reagent or mixture of reagents capable of removing a protecting group selected from the group consisting of t-butyl, benzyl, 2-chlorotrityl (2-Cl-Trt), 2,4-dimethoxybenzyl (Dmb), allyl, 1,1-dimethylallyl (Dma), and p-nitrobenzyl (pNB).

[0104] 90. The method of any one of clauses 52 to 89, wherein the third deprotecting agent in step (g) is selected from the group consisting of trifluoroacetic acid (TFA), hydrofluoric acid (HF), triflic acid (TfOH), trifluoroacetic acid (TFA) / water / triisopropylsilane mixture, trifluoroacetic acid (TFA) / dichloromethane (DCM) mixture, triethylsaline (TES), indole, phenol / anisole mixture, and thioanisole.

[0105] 91. The method of any one of clauses 52 to 90, wherein the third deprotecting agent in step (g) is a trifluoroacetic acid (TFA) / water / triisopropylsilane mixture.

[0106] 92. The method of any one of clauses 52 to 91, wherein the cleavage reagent in step (g) is trifluoroacetic acid (TFA) or a trifluoroacetic acid (TFA) / water / triisopropylsilane mixture.

[0107] 93. The method of any one of clauses 52 to 92, wherein the cleavage reagent in step (g) is a trifluoroacetic acid (TFA) / water / triisopropylsilane mixture.

[0108] 94. The method of any one of clauses 52-93, wherein the organic solvent in step (g) is selected from the group consisting of CH2Cl2, THF, diethyl ether, MeOH, EtOH, acetone, and acetonitrile.

[0109] 95. Prior to contacting with the resin-based compound of formula 5, a compound of formula 6, a solvent, a coupling agent, and optionally a base and / or additives are combined to form a compound of formula 6' [ka] The method of any one of clauses 52-94, comprising forming a mixture containing an activated compound of the formula:

[0110] 96. Formula 6' [ka] The method of any one of clauses 52 to 94, wherein the compound of the resin-based compound of formula 5, the compound of formula 6, the solvent, the coupling agent, and optionally the base and / or the additive are contacted together prior to or simultaneously with the in situ formation of the activated compound of formula 6', wherein LG of compound 6' is a leaving group.

[0111] 97. LG of compound 6' is 1-(λ 1 -oxidanyl)-1H-benzo[d][1,2,3]triazole, 3-(λ 1 -Oxydanyl)-3H-[1,2,3]triazolo[4,5-b]pyridine, ethyl cyano(hydroxyimino)acetate (Oxyma), bromide, 3-(λ 1 -Oxidaneyl)benzo[d][1,2,3]triazin-4(3H)-one, 6-chloro-1-(λ 1 -Oxidaneyl)-1H-benzo[d][1,2,3]triazole, 2,4-dimethoxy-6-(lambda 1-Oxidaneyl)-1,3,5-triazine, ethabonic acid, propylphosphonic acid, dicyclohexylurea, 1-(3-(dimethyl-λ 4 97. The method of claim 95 or 96, wherein the aryl group is selected from the group consisting of -azanyl)propyl)-3-ethylurea, and imidazolyl.

[0112] 98. The method of any one of clauses 95 to 97, wherein LG of compound 6' is imidazolyl.

[0113] 99. Prior to contacting with the resin-based compound of formula H, a compound of formula 9, a solvent, a coupling agent, and optionally a base and / or additives are combined to form a compound of formula 9' [ka] The method of any one of the preceding claims, forming a mixture containing an activated compound of formula: wherein LG of compound 9' is a leaving group.

[0114] 100. Formula 9' [ka] The method of any one of clauses 52 to 98, wherein the resin-based compound of formula 8, the compound of formula 9, the solvent, the coupling agent, and optionally the base and / or additive are contacted together prior to or simultaneously with the in situ formation of the activated compound of formula (wherein LG of compound 9' is a leaving group).

[0115] 101. LG of compound 9' is 1-(λ 1 -oxidanyl)-1H-benzo[d][1,2,3]triazole, 3-(λ 1 -Oxydanyl)-3H-[1,2,3]triazolo[4,5-b]pyridine, ethyl cyano(hydroxyimino)acetate (Oxyma), bromide, 3-(λ 1 -Oxidaneyl)benzo[d][1,2,3]triazin-4(3H)-one, 6-chloro-1-(λ 1-Oxidaneyl)-1H-benzo[d][1,2,3]triazole, 2,4-dimethoxy-6-(lambda 1 -Oxidaneyl)-1,3,5-triazine, ethabonic acid, propylphosphonic acid, dicyclohexylurea, 1-(3-(dimethyl-λ 4 101. The method of claim 99 or 100, wherein the aryl group is selected from the group consisting of -azanyl)propyl)-3-ethylurea, and imidazolyl.

[0116] 102. The method of any one of clauses 99 to 101, wherein LG of compound 9' is imidazolyl.

[0117] 103. Formula 4 [ka] (In the formula, PG 1 is an amine protecting group, PG 2 and P.G. 3 are each independently a carboxyl protecting group. Resin-based compound.

[0118] 104. Formula 5 [ka] (In the formula, PG 2 and P.G. 3 are each independently a carboxyl protecting group. Resin-based compound.

[0119] 105. Formula 7 [ka] (In the formula, PG 2 and P.G. 3 are each independently a carboxy protecting group, and PG 4 is an amine protecting group) Resin-based compound.

[0120] 106. Formula 8 [ka] (In the formula, PG 2 and P.G. 3 are each independently a carboxy protecting group. Resin-based compound.

[0121] 107. Formula 10 [ka] (In the formula, each PG 2 , P.G. 3 , and P.G. 5 are independently carboxyl protecting groups. Resin-based compound.

[0122] 108. PG 1 and P.G. 4 108. The resin based compound of any one of clauses 103-107, wherein, if present, is independently selected from the group consisting of 9-fluorenylmethyl-carbonyl (Fmoc), benzyl carbamate (Cbz), t-butyloxycarbonyl (Boc), acetamide, trifluoroacetamide, p-toluenesulfonamide, triphemylmethyl, monomethoxytrityl (MMt), and 4-methyltrityl (Mtt).

[0123] 109. PG 1 109. The resinous compound of any one of clauses 103 to 108, wherein, if present, is monomethoxytrityl (MMt) or 4-methyltrityl (Mtt).

[0124] 110. PG 1 101. The resinous compound of any one of clauses 103 to 109, wherein, if present, is 4-methyltrityl (Mtt).

[0125] 111. PG 4111. The resinous compound according to any one of clauses 103 to 110, wherein, if present, is 9-fluorenylmethyl-carbonyl.

[0126] 112. P.G. 2 , P.G. 3 , and P.G. 5 and, if present, each is t-butyl.

[0127] 113. Formula C [ka] (In the formula, PG 2 is an amine protecting group, and each R 1 is C1 to C4 alkyl or cyclohexyl).

[0128] 114. Formula C' [ka] (In the formula, each R 1 is C1-C4 alkyl or cyclohexyl) Resin-based compound.

[0129] 115. Formula D [ka] (In the formula, PG 1 is an amine protecting group, and each R 1 is C1-C4 alkyl or cyclohexyl) Resin-based compound.

[0130] 116. Formula D' [ka] (In the formula, each R 1 is C1-C4 alkyl or cyclohexyl) Resin-based compound.

[0131] 117. Formula E [ka] (In the formula, PG 1 is an amine protecting group, and each R 1 is C1-C4 alkyl or cyclohexyl) Resin-based compound.

[0132] 118. Formula E' [ka] (In the formula, each R 1 is C1-C4 alkyl or cyclohexyl) Resin-based compound.

[0133] 119. Formula F [ka] (In the formula, each R 1 is C1-C4 alkyl or cyclohexyl) Resin-based compound.

[0134] 120. P.G. 1 118. The resin-based compound according to claim 115 or 117, wherein is 9-fluorenylmethyl-carbonyl (Fmoc) or t-butylcarbonyl (Boc).

[0135] 121. P.G. 1 121. The resin based compound according to any one of clauses 115, 117 or 120, wherein is 9-fluorenylmethyl-carbonyl (Fmoc).

[0136] 122. P.G. 2 114. The resin-based compound according to claim 113, wherein is monomethoxytrityl (MMt) or 4-methyltrityl (Mtt).

[0137] 123. PG 2 123. The resin-based compound according to claim 113 or 122, wherein is 4-methyltrityl (Mtt).

[0138] 124. R 1 Item 113. The resin compound according to any one of items 113 to 119, wherein is t-butyl.

[0139] definition As used herein, the term "alkyl" includes a chain of carbon atoms, which is optionally branched and contains from 1 to 4 carbon atoms, etc., and may be referred to as a "lower alkyl." Exemplary alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl.

[0140] As used herein, unless otherwise indicated, the term "abnormal cell growth" refers to cell growth that is independent of normal regulatory mechanisms (eg, loss of contact inhibition).

[0141] The term "subject" refers to a mammalian patient in need of such treatment, such as a human.

[0142] As used herein, the term "protecting group" or "PG" refers to any group generally known to one skilled in the art that can be introduced into a molecule by chemical modification of a functional group, such as an amine or hydroxyl, to obtain chemoselectivity in a subsequent chemical reaction. It will be appreciated that such protecting groups can then be removed from the functional group at a later point in the synthesis to provide further opportunities for reaction at such functional group, or to unmask such functional group in the case of the final product. Protecting groups are described, for example, in Wuts, PGM, Greene, TW, Greene, TW, & John Wiley & Sons. (2006). Greene's protective groups in organic synthesis. Hoboken, NJ: Wiley-Intersciences. One skilled in the art will readily recognize chemical process conditions under which such protecting groups can be introduced onto functional groups. Suitable amine protecting groups useful in the context of the present disclosure include, but are not limited to, 9-fluorenylmethyl-carbonyl (Fmoc) and t-butylcarbonyl (Boc).

[0143] As used herein, the term "leaving group" of "LG" refers to any group commonly known to those skilled in the art that leaves with a pair of electrons in a heterolytic bond cleavage. The leaving group can be an anion or a neutral molecule, but in either case, it is important that the leaving group is capable of stabilizing the additional electron density that results from the heterolytic bond cleavage. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0144] Before the present disclosure is further described, it is to be understood that this disclosure is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present disclosure will be limited only by the appended claims.

[0145] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents, applications, published applications and other publications referenced herein are incorporated by reference in their entirety. If a definition set forth in this section contradicts or otherwise conflicts with a definition set forth in a patent, application, or other publication incorporated herein by reference, the definition set forth in this section shall take precedence over the definition incorporated herein by reference.

[0146] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It is further noted that the claims may be drafted to exclude any optional element. Thus, this statement is intended to serve as a prelude for using exclusive terminology, such as "sole," "only," and the like, in connection with reciting claim elements or using a "negative" limitation.

[0147] Synthesis method Method 1: In some embodiments, the present disclosure provides a compound of formula I [ka] The present invention provides a method for preparing a compound of the formula: Formula A [ka] contacting the resin-based compound of formula A' with a first deprotecting agent in a polar aprotic solvent to produce a compound of formula A' [ka] or providing a resin-based compound of b. A resin-based compound of formula A' is mixed with a resin-based compound of formula B [ka] in the presence of an organic solvent and a base. [ka] or providing a resin-based compound of formula C c. contacting the resin-based compound of formula C with a second deprotecting agent in the presence of an organic solvent to produce a compound of formula C' [ka] or providing a resin-based compound of d. The resin-based compound of formula C' is reacted with PG in the presence of a polar aprotic solvent, a coupling agent, and a base. 1 -3-(2-naphthyl)-L-alanine to form a compound of formula D [ka] or providing a resin-based compound of e. contacting the resin-based compound of formula D with a first deprotecting agent in a polar aprotic solvent to produce a compound of formula D' [ka] or providing a resin-based compound of f. The resin-based compound of formula D' is reacted with PG in the presence of a polar aprotic solvent, a coupling agent, and a base. 1 - contacting with tranexamic acid to form a compound of formula E [ka] or providing a resin-based compound of g. contacting the resin-based compound of formula E with a first deprotecting agent in a polar aprotic solvent to produce a compound of formula E' [ka] or providing a resin-based compound of h. Resin-based compound of formula E' with a compound of formula E 2’ [ka] in the presence of a polar aprotic solvent, a coupling agent, and a base to obtain a compound of formula F [ka] or providing a resin-based compound of i. contacting the resin-based compound of formula F with a third deprotecting and / or cleaving reagent in an organic solvent; Including, In the ceremony, P.G. 1 and P.G. 2 is an amine protecting group, LG is a leaving group, and R 1 is C1-C4 alkyl or cyclohexyl.

[0148] Method 2: Alternatively, in some embodiments, the present disclosure provides a compound represented by formula 1 [ka] The present invention provides a method for preparing a compound of the formula: a. Formula 1 [ka] contacting the resin-based compound of formula 2 with a first deprotecting agent in the presence of a solvent to form a compound of formula 2 [ka] or providing a resin-based compound of b. The resin-based compound of formula 2 is mixed with a compound of formula 3 [ka] in the presence of an organic solvent and a base to form a compound of formula 4 [ka] or providing a resin-based compound of c. contacting the resin-based compound of formula 4 with a second deprotecting agent in the presence of an organic solvent to produce a compound of formula 5 [ka] or providing a resin-based compound of d. The resin-based compound of formula 5 is reacted with a compound of formula 6 [ka] in the presence of a solvent, a coupling agent, and optionally a base and / or an additive to obtain a compound of formula 7 [ka] or providing a resin-based compound of e. contacting the resin-based compound of formula 7 with a third deprotecting agent in the presence of a solvent to produce a compound of formula 8. [ka] or providing a resin-based compound of f. The resin-based compound of formula 8 is reacted with a compound of formula 9 [ka] in the presence of a solvent, a coupling agent, and optionally a base and / or an additive to obtain a compound of formula 10 [ka] or providing a resin-based compound of g. contacting the resin-based compound of formula 10 with at least one third deprotecting and / or cleaving agent in the presence of an organic solvent to provide a compound of formula I. one or more of In the formula, PG, PG 1 and P.G. 4 is an amine protecting group, PG 2 , P.G. 3 , and P.G. 5 is a carboxyl protecting group, and LG of compound C is a leaving group.

[0149] The present disclosure provides a method for preparing a compound of formula I described in the above paragraph (by method 1 or method 2), and it will be recognized that the method alternatively includes more than one of the steps listed. Thus, the present disclosure provides a method for preparing a compound of formula I, comprising steps (a) and (b). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (b) and (c). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (c) and (d). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (d) and (e). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (e) and (f). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (f) and (g). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (g) and (h). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (h) and (i). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (a), (b) and (c). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (b), (c) and (d). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (c), (d) and (e). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (d), (e) and (f). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (e), (f) and (g). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (d), (e) and (f). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (f), (g) and (h). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (d), (e) and (f). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (g), (h) and (i). Alternatively, the present disclosure provides a method for preparing a compound of formula I, comprising steps (a), (b), (c), (d), (e), (f), (g) and (h).

[0150] In various embodiments described herein, it will be recognized by those skilled in the art that the selection of the protecting group used in the preparation of compound (I) can be selected from various alternatives known in the art. Furthermore, it will be recognized that any protecting group scheme is selected such that the protecting groups used are an orthogonal protection strategy. As used herein, "orthogonal protection" refers to a strategy that allows the deprotection of one or more protecting groups among multiple protecting groups, where the protecting groups are removed one at a time, each without affecting the others, using a dedicated set of reaction conditions, or are removed together as a subset of all protecting groups used in the synthesis, without affecting the others, using a dedicated set of reaction conditions.

[0151] In step (a) of method 1, PG on a resin-based compound of formula A 1 can be a protecting group such as 9-fluorenylmethyl-carbonyl (Fmoc) or t-butylcarbonyl (Boc), and PG 2 can be an orthogonal protecting group such as 4-methyltrityl (Mtt) or monomethoxytrityl (MMt). The first deprotecting agent in step (a) of method 1 is PG 1 and / or PG 2The first deprotecting agent may be an acidic or basic deprotecting agent, depending on the identity of the first deprotecting agent. In particular, the first deprotecting agent may be a basic deprotecting agent, such as piperidine, morpholine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), diethylamine (DEA), dicyclohexamine, 4-methylpiperidine (4MP), tris(2-aminoethyl)amine, pyridine, or collidine. In particular, the first deprotecting agent may be an acidic deprotecting agent, such as TFA, HCl, or H3PO4. Suitable deprotecting agents and conditions for step (a) of method 1 can be found, for example, in Wuts, PGM, Greene, TW, Greene, TW, & John Wiley & Sons. (2006). Greene's protective groups in organic synthesis. Hoboken, NJ: Wiley-Interscience. In step (a) of method 1, the base used to remove the Fmoc group is piperidine. The polar aprotic solvent in step (a) of method 1 can be any polar aprotic solvent known in the art. Suitable polar aprotic solvents include, but are not limited to, dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO). It will be appreciated that step (a) of method 1 can be carried out at any temperature commonly used in solid phase synthesis, such as at room temperature, under cooling, or under warm conditions. The deprotection step (a) of method 1 can be carried out for a period of time, such as about 5 minutes to about 200 minutes, or about 10 minutes to about 100 minutes, or about 40 minutes to about 60 minutes to about 15 minutes, or about 30 minutes, or about 90 minutes. Following the deprotection step (a) of Method 1, the resin-based compound of Formula A' prepared in step (a) may be washed with reagents commonly known in the art, such as DMF, IPA, CH2Cl2, and the like.

[0152] In step (b) of method 1, R on the compound of formula B 1R may be a C1-C4 alkyl, preferably tert-butyl, or cyclohexyl, and LG is a leaving group, such as an active ester. Suitable active esters known in the art include, but are not limited to, 4-nitrophenyl, 2,4,5-trichlorophenyl, NHS ester, benzotriazyl, and the like. In some embodiments, LG is a 4-nitrophenyl ester. R 1 The choice is PG 1 , P.G. 2 It will be appreciated that the choice of urea formation reaction and / or resin bead attachment is orthogonal. In step (b) of method 1, the base can be any base commonly known in the art for urea formation reactions. Suitable bases include N,N-diisopropylethylamine ( i Examples of suitable solvents include, but are not limited to, Pr2NEt), triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine. Step (b) of method 1 can be carried out in a solvent, such as an organic solvent, which can be any solvent commonly known in the art for resin-based synthesis. Suitable organic solvents include, but are not limited to, CHCl2, THF, diethyl ether, MeOH, EtOH, acetone, acetonitrile, and the like. In some embodiments, the solvent is CHCl2. It will be appreciated that step (b) of method 1 can be carried out at any temperature commonly used in solid phase synthesis, such as at room temperature, under cooling, or under warming conditions. The deprotection step (b) of method 1 can be carried out for a period of time, such as about 5 minutes to about 200 minutes, or about 10 minutes to about 100 minutes, or about 40 minutes to about 60 minutes to about 15 minutes, or about 30 minutes, or about 90 minutes. Following the deprotection step (a) of Method 1, the resin-based compound of Formula C prepared in step (b) may be washed with reagents commonly known in the art, such as DMF, IPA, CH2Cl2, and the like.

[0153] In step (c) of method 1, the second deprotecting agent is PG 1 and / or PG 2Depending on the identity of the second deprotection agent, the second deprotection agent may be an acidic deprotection agent, a basic deprotection agent, or other deprotection agent. In some embodiments, the second deprotection agent may be an acidic deprotection agent. Suitable acidic deprotection agents include, but are not limited to, TFA, HCl, or H3PO4. In some embodiments, the second deprotection agent in step (c) of method 1 is selected from the group consisting of trifluoroacetic acid (TFA), acetic acid, trifluoroethanol, hexafluoroisopropanol, dichloromethane, and combinations thereof. In some embodiments, the second deprotection agent is a PG 2 is TFA, such as when is Mtt. In some embodiments, the second deprotection is 2 In some embodiments, the second deprotecting agent is a mixture of acetic acid and trifluoroethanol, such as when PG is Mtt. 2 may be a palladium deprotecting agent, such as when is alloc. Suitable deprotecting agents and conditions for step (c) of method 1 can be found, for example, in Wuts, PGM, Greene, TW, Greene, TW, & John Wiley & Sons. (2006). Greene's protective groups in organic synthesis. Hoboken, NJ: Wiley-Interscience. An acidic deprotecting agent (TFA) is used to cleave Mtt. Other suitable acidic deprotecting agents include, but are not limited to, acetic acid, trifluoroethanol, dichloromethane, hexafluoroisopropanol, dichloromethane, and combinations thereof. Step (c) of method 1 may be carried out in a solvent, such as an organic solvent. The organic solvent may be any solvent generally known in the art for resin-based synthesis. Suitable organic solvents include, but are not limited to, CH2Cl2, THF, diethyl ether, MeOH, EtOH, acetone, acetonitrile, and the like. In some embodiments, the solvent is CH2Cl2.

[0154] In step (d), PG 1 -PG on 3-(2-naphthyl)-L-alanine 1 [ka] may be a protecting group such as 9-fluorenylmethyl-carbonyl (Fmoc) or t-butylcarbonyl (Boc). 1 For the selection of, see step (a) of method 1 above. In step (d) of method 1, the polar aprotic solvent can be any polar aprotic solvent commonly known in the art. Suitable polar aprotic solvents include, but are not limited to, N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO). In some embodiments, the polar aprotic solvent is DMF. Suitable bases include, but are not limited to, triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine. In some embodiments, the base in step (d) of method 1 is N,N-diisopropylethylamine ( i The coupling agent is (Pr2NEt). The coupling agent can be any coupling agent commonly known in the art for resin-based synthesis. Suitable coupling agents include, but are not limited to, HATU, TBTU, HBTU, CDMT, HDMC, COMU, TATU, TOTT, T3P, DMTMM. In some embodiments, the coupling agent in step (d) of method 1 is (PyBOP). It will be appreciated that step (d) of method 1 can be carried out at any temperature commonly used in solid phase synthesis, such as at room temperature, under cooling, or under warm conditions. The coupling step (d) of method 1 can be carried out for a period of time such as about 5 minutes to about 200 minutes, or about 10 minutes to about 100 minutes, or about 40 minutes to about 60 minutes to about 15 minutes, or about 30 minutes, or about 90 minutes. Following coupling step (d) of method 1, the resin-based compound of formula D produced in step (d) of method 1 can be washed with a reagent commonly known in the art, such as DMF, IPA, CH2Cl2, etc.

[0155] In step (e) of method 1, PG on a resin-based compound of formula D 1 can be a protecting group such as 9-fluorenylmethyl-carbonyl (Fmoc) or t-butylcarbonyl (Boc), and PG 2 can be an orthogonal protecting group such as 4-methyltrityl (Mtt) or monomethoxytrityl (MMt). The first deprotecting agent in step (e) of method 1 is PG 1 and / or PG 2The first deprotecting agent may be an acidic or basic deprotecting agent, depending on the identity of the first deprotecting agent. In particular, the first deprotecting agent may be a basic deprotecting agent, such as piperidine, morpholine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), diethylamine (DEA), dicyclohexamine, 4-methylpiperidine (4MP), tris(2-aminoethyl)amine, pyridine, or collidine. In particular, the first deprotecting agent may be an acidic deprotecting agent, such as TFA, HCl, or H3PO4. Suitable deprotecting agents and conditions for step (e) of method 1 can be found, for example, in Wuts, PGM, Greene, TW, Greene, TW, & John Wiley & Sons. (2006). Greene's protective groups in organic synthesis. Hoboken, NJ: Wiley-Interscience. The polar aprotic solvent of step (e) of method 1 may be any polar aprotic solvent known in the art. Suitable polar aprotic solvents include, but are not limited to, dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO). It will be appreciated that step (e) of method 1 can be carried out at any temperature commonly used in solid phase synthesis, such as at room temperature, under cooling, or under warm conditions. The deprotection step (e) of method 1 can be carried out for a period of time, such as about 5 minutes to about 200 minutes, or about 10 minutes to about 100 minutes, or about 40 minutes to about 60 minutes to about 45 minutes, or about 60 minutes, or about 90 minutes. Following the deprotection step (e) of method 1, the resin-based compound of formula D' prepared in step (e) of method 1 can be washed with a reagent commonly known in the art, such as DMF, IPA, CHCl.

[0156] In step (f) of the first method, PG 1 -PG on tranexamic acid reagent 1 teeth, [ka] It may be a protecting group such as 9-fluorenylmethyl-carbonyl (Fmoc) or t-butylcarbonyl (Boc). 1 For the selection of, see step (a) of method 1 above. In step (f) of method 1, the polar aprotic solvent can be any polar aprotic solvent commonly known in the art. Suitable polar aprotic solvents include, but are not limited to, N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO). In some embodiments, the polar aprotic solvent is DMF. Suitable bases include, but are not limited to, triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine. In some embodiments, the base in step (f) of method 1 is N,N-diisopropylethylamine ( i The coupling agent is (Pr2NEt). The coupling agent can be any coupling agent commonly known in the art for resin-based synthesis. Suitable coupling agents include, but are not limited to, HATU, TBTU, HBTU, CDMT, HDMC, COMU, TATU, TOTT, T3P, DMTMM. In some embodiments, the coupling agent in step (f) of method 1 is (PyBOP). It will be appreciated that step (f) of method 1 can be carried out at any temperature commonly used in solid phase synthesis, such as at room temperature, under cooling, or under warm conditions. The coupling step (f) of method 1 can be carried out for a period of time such as about 5 minutes to about 200 minutes, or about 10 minutes to about 100 minutes, or about 40 minutes to about 60 minutes to about 15 minutes, or about 30 minutes, or about 90 minutes. Following coupling step (f) of method 1, the resin-based compound of formula E produced in step (f) of method 1 can be washed with a reagent commonly known in the art, such as DMF, IPA, CH2Cl2, etc.

[0157] In step (g) of method 1, PG on the resin-based compound of formula E1 can be a protecting group such as 9-fluorenylmethyl-carbonyl (Fmoc) or t-butylcarbonyl (Boc), and PG 2 can be an orthogonal protecting group such as 4-methyltrityl (Mtt) or monomethoxytrityl (MMt). The first deprotecting agent in step (g) of method 1 is PG 1 and / or PG 2The first deprotecting agent may be an acidic or basic deprotecting agent, depending on the identity of the first deprotecting agent. In particular, the first deprotecting agent may be a basic deprotecting agent, such as piperidine, morpholine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), diethylamine (DEA), dicyclohexamine, 4-methylpiperidine (4MP), tris(2-aminoethyl)amine, pyridine, or collidine. In particular, the first deprotecting agent may be an acidic deprotecting agent, such as TFA, HCl, or H3PO4. Suitable deprotecting agents and conditions for step (g) of method 1 can be found, for example, in Wuts, PGM, Greene, TW, Greene, TW, & John Wiley & Sons. (2006). Greene's protective groups in organic synthesis. Hoboken, NJ: Wiley-Interscience. The polar aprotic solvent of step (g) of method 1 may be any polar aprotic solvent known in the art. Suitable polar aprotic solvents include, but are not limited to, dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO). It will be appreciated that step (g) of method 1 can be carried out at any temperature commonly used in solid phase synthesis, such as at room temperature, under cooling, or under warm conditions. The deprotection step (g) of method 1 can be carried out for a period of time such as about 5 minutes to about 200 minutes, or about 10 minutes to about 100 minutes, or about 40 minutes to about 60 minutes to about 45 minutes, or about 60 minutes, or about 90 minutes. Following the deprotection step (g) of method 1, the resin-based compound of formula E' prepared in step (g) of method 1 can be washed with a reagent commonly known in the art, such as DMF, IPA, CHCl.

[0158] In step (h) of method 1, the polar aprotic solvent can be any polar aprotic solvent commonly known in the art. Suitable polar aprotic solvents include, but are not limited to, N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO). In some embodiments, the polar aprotic solvent is DMF. Suitable bases include, but are not limited to, triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine. In some embodiments, the base in step (h) of method 1 is N,N-diisopropylethylamine ( i The coupling agent is (Pr2NEt). The coupling agent can be any coupling agent commonly known in the art for resin-based synthesis. Suitable coupling agents include, but are not limited to, HATU, TBTU, HBTU, CDMT, HDMC, COMU, TATU, TOTT, T3P, DMTMM. In some embodiments, the coupling agent in step (h) of method 1 is (PyBOP). It will be appreciated that step (h) of method 1 can be carried out at any temperature commonly used in solid phase synthesis, such as at room temperature, under cooling, or under warm conditions. The coupling step (h) of method 1 can be carried out for a period of time such as about 5 minutes to about 200 minutes, or about 10 minutes to about 100 minutes, or about 40 minutes to about 60 minutes to about 15 minutes, or about 30 minutes, or about 90 minutes. Following coupling step (h) of method 1, the resin-based compound of formula F produced in step (h) of method 1 can be washed with a reagent commonly known in the art, such as DMF, IPA, CH2Cl2, etc.

[0159] In step (i) of method 1, the third deprotecting agent may be the same as or different from the cleavage reagent. In some embodiments, the third deprotecting agent is, for example, R 1 may be HF or TfOH, such as when R is cyclohexyl and the cleavage reagent may be TFA. 1The third deprotecting agent and cleaving reagent can both be TFA, such as when is tert-butyl. In some embodiments, the third deprotecting agent and cleaving reagent can both be a mixture of TFA, water, and triisopropylsilane. Other suitable deprotecting agents include, but are not limited to, trifluoroacetic acid (TFA) / dichloromethane (DCM) mixtures, triethylsaline (TES), indole, phenol / anisole mixtures, and thioanisole. Step (i) of method 1 can be carried out in a solvent, such as an organic solvent. The organic solvent can be any solvent generally known in the art for resin-based synthesis. Suitable organic solvents include, but are not limited to, CH2Cl2, THF, diethyl ether, MeOH, EtOH, acetone, acetonitrile, and the like. In some embodiments, the solvent is CH2Cl2.

[0160] In step (a) of method 2, PG on the resin-based compound of formula 1 can be a protecting group, particularly an amine protecting group such as 9-fluorenylmethyl-carbonyl (Fmoc), benzyl carbamate (Cbz), t-butyloxycarbonyl (Boc), acetamide, trifluoroacetamide, p-toluenesulfonamide, triphemylmethyl, monomethoxytrityl (MMt), or 4-methyltrityl (Mtt). Also, in step (a) of method 2, PG can be a protecting group, particularly an amine protecting group such as 9-fluorenylmethyl-carbonyl (Fmoc), benzyl carbamate (Cbz), t-butyloxycarbonyl (Boc), acetamide, trifluoroacetamide, p-toluenesulfonamide, triphemylmethyl, monomethoxytrityl (MMt), or 4-methyltrityl (Mtt). 1 can be a protecting group, particularly an amine protecting group such as 9-fluorenylmethyl-carbonyl (Fmoc), benzyl carbamate (Cbz), t-butyloxycarbonyl (Boc), acetamide, trifluoroacetamide, p-toluenesulfonamide, triphemylmethyl, monomethoxytrityl (MMt), or 4-methyltrityl (Mtt), which is orthogonal to PG. In some embodiments, PG can be 9-fluorenylmethyl-carbonyl (Fmoc) or t-butylcarbonyl (Boc), and PG can be 10-methyltrityl (Mtt), 15-methyltrityl (Mtt), 16-methyltrityl (Mtt), 17-methyltrityl (Mtt), 18-methyltrityl (Mtt), 19-methyltrityl (Mtt), 20-methyltrityl (Mtt), 21-methyltrityl (Mtt), 22-methyltrityl (Mtt), 23-methyltrityl (Mtt), 24-methyltrityl (Mtt), 25-methyltrityl (Mtt), 26-methyltrityl (Mtt), 27-methyltrityl (Mtt), 28-methyltrityl (Mtt), 29-methyltrityl (Mtt), 30-methyltrityl (Mtt), 31-methyltrityl (Mtt), 32-methyltrityl (Mtt), 33-methyltrityl (Mtt), 34-methyltrityl (Mtt), 35-methyltrityl (Mtt), 36-methyltrityl (Mtt), 37-methyltrityl (Mtt), 38-methyltrityl (Mtt), 39-methyltrityl (Mtt), 38-methyltrityl (Mtt), 39-methyltrityl (Mtt), 32-methyltrityl (Mtt), 34-methyltrityl (Mtt), 35-methyltrit 1 may be an orthogonal protecting group such as 4-methyltrityl (Mtt) or monomethoxytrityl (MMt).

[0161] The first deprotecting agent in step (a) of method 2 can be a reagent or mixture of reagents capable of removing an amine protecting group, such as 9-fluorenylmethyl-carbonyl (Fmoc), benzyl carbamate (Cbz), t-butyloxycarbonyl (Boc), acetamide, trifluoroacetamide, p-toluenesulfonamide, triphemylmethyl, monomethoxytrityl (MMt), or 4-methyltrityl (Mtt). In some embodiments, the first deprotecting agent in step (a) of method 2 comprises a reagent or mixture of reagents capable of removing 9-fluorenylmethyl-carbonyl. In some embodiments, the first deprotecting agent in step (a) of method 2 comprises PG and / or PG 1 The first deprotecting agent may be an acidic or basic deprotecting agent, depending on the identity of the first deprotecting agent. In particular, the first deprotecting agent may be a basic deprotecting agent, such as piperidine, morpholine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), diethylamine (DEA), dicyclohexamine, 4-methylpiperidine (4MP), tris(2-aminoethyl)amine, pyridine, or collidine. In particular, the first deprotecting agent may be an acidic deprotecting agent, such as TFA, HCl, or H3PO4. Suitable deprotecting agents and conditions for step (a) of method 2 can be found, for example, in Wuts, PGM, Greene, TW, Greene, TW, & John Wiley & Sons. (2006). Greene's protective groups in organic synthesis. Hoboken, NJ: Wiley-Interscience.

[0162] In some embodiments of step (a) of method 2, the base used to remove the Fmoc group can be an amine base, such as piperidine. In some embodiments, the polar aprotic solvent of step (a) of method 2 can be any polar aprotic solvent known in the art. Suitable polar aprotic solvents include, but are not limited to, dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO). It will be appreciated that step (a) can be carried out at any temperature commonly used in solid phase synthesis, such as room temperature, under cooling, or under warm conditions. The deprotection step (a) of Method 2 can be carried out for a period of time such as about 5 minutes to about 200 minutes, or about 10 minutes to about 100 minutes, or about 40 minutes to about 60 minutes to about 15 minutes, or about 30 minutes, or about 90 minutes, etc. Following the deprotection step (a) of Method 2, the resin-based compound of Formula B prepared in step (a) of Method 2 can be washed with a reagent commonly known in the art such as DMF, IPA, CH2Cl2, etc.

[0163] In step (b) of method 2, PG on the compound of formula 3 2 and / or PG 3 may each independently be a carboxy protecting group, and LG is a leaving group, such as an active ester. Suitable active esters known in the art include, but are not limited to, imidazolyl, 4-nitrophenoxy, phenoxy, 2,4,5-trichlorophenyl, NHS ester, benzotriazyl, and the like. In some embodiments, LG of compound 3 is selected from imidazolyl, 4-nitrophenoxy, and phenoxy. In some embodiments, LG of compound 3 is a 4-nitrophenyl ester. PG 2 and / or PG 3 The choice is PG and PG 1 It will be appreciated that the selection of PG is orthogonal. 2 and / or PG 3In step (b) of method 2, the base can be any base commonly known in the art for urea-forming reactions. Suitable bases include N,N-diisopropylethylamine ( i Examples of suitable amines include, but are not limited to, PrNEt), triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine.

[0164] Step (b) of method 2 can be carried out in a solvent, such as an organic solvent. The organic solvent can be any solvent commonly known in the art for resin-based synthesis. Suitable organic solvents include, but are not limited to, CH2Cl2, THF, diethyl ether, MeOH, EtOH, acetone, acetonitrile, and the like. In some embodiments, the solvent is CH2Cl2. It will be appreciated that step (b) of method 2 can be carried out at any temperature commonly used in solid phase synthesis, such as at room temperature, under cooling, or under warm conditions. The deprotection step (b) of method 2 can be carried out for a period of time, such as about 5 minutes to about 200 minutes, or about 10 minutes to about 100 minutes, or about 40 minutes to about 60 minutes to about 15 minutes, or about 30 minutes, or about 90 minutes. Following step (b) of method 2, the resin-based compound of formula 4 produced in step (b) can be washed with a reagent commonly known in the art, such as DMF, IPA, CH2Cl2, and the like.

[0165] In step (c) of method 2, the second deprotecting agent can be a reagent or mixture of reagents capable of removing a protecting group selected from the group consisting of 9-fluorenylmethyl-carbonyl (Fmoc), benzyl carbamate (Cbz), t-butyloxycarbonyl (Boc), acetamide, trifluoroacetamide, p-toluenesulfonamide, triphemylmethyl, monomethoxytrityl (MMt), and 4-methyltrityl (Mtt). In some embodiments, the second deprotecting agent can be a reagent or mixture of reagents capable of removing 4-methyltrityl (Mtt). In some embodiments, the second deprotecting agent is PG 1 , P.G. 2and / or PG 3 Depending on the identity of the second deprotection agent, the second deprotection agent may be an acidic deprotection agent, a basic deprotection agent, or other deprotection agent. In some embodiments, the second deprotection agent may be an acidic deprotection agent. Suitable acidic deprotection agents include, but are not limited to, TFA, HCl, or H3PO4. In some embodiments, the second deprotection agent in step (c) of method 2 is selected from the group consisting of trifluoroacetic acid (TFA), acetic acid, trifluoroethanol, hexafluoroisopropanol, and combinations thereof. In some embodiments, the second deprotection agent is selected from the group consisting of PG 2 In some embodiments, the second deprotection in step (c) of method 2 is TFA, such as when PG 2 is a mixture of acetic acid and trifluoroethanol, such as when Mtt is. Suitable deprotection agents and conditions for step (c) of method 2 can be found, for example, in Wuts, PGM, Greene, TW, Greene, TW, & John Wiley & Sons. (2006). Greene's protective groups in organic synthesis. Hoboken, NJ: Wiley-Interscience. Step (c) of method 2 can be carried out in a solvent, such as an organic solvent. The organic solvent can be any solvent commonly known in the art for resin-based synthesis. Suitable organic solvents include, but are not limited to, CH2Cl2, THF, diethyl ether, MeOH, EtOH, acetone, acetonitrile, and the like. In some embodiments, the solvent is CH2Cl2. Following the deprotection step (c) of method 2, the resin-based compound of formula 5 produced in step (c) of method 2 can be washed with reagents commonly known in the art, such as DMF, IPA, CH2Cl2, and the like.

[0166] In step (d) of method 2, PG 4can be a protecting group, particularly an amine protecting group, 9-fluorenylmethyl-carbonyl (Fmoc), benzyl carbamate (Cbz), t-butyloxycarbonyl (Boc), acetamide, trifluoroacetamide, p-toluenesulfonamide, triphemylmethyl, monomethoxytrityl (MMt), and 4-methyltrityl (Mtt). In some embodiments, PG 4 can be 9-fluorenylmethyl-carbonyl (Fmoc) or t-butylcarbonyl (Boc). 4 For the selection of, see step (a) of method 2 above. In step (d) of method 2, the solvent can be a polar aprotic solvent. In some embodiments, the polar aprotic solvent can be any polar aprotic solvent commonly known in the art. Suitable polar aprotic solvents include, but are not limited to, dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO). In some embodiments, the polar aprotic solvent is DMF. Suitable bases for use in step (d) of method 2 include N,N-diisopropylethylamine ( i In some embodiments, the base in step (d) includes, but is not limited to, N,N-diisopropylethylamine (N ... i PrNEt).

[0167] The coupling agent may be any coupling agent commonly known in the art for resin-based synthesis. Suitable coupling agents for use in step (d) of method 2 include benzotriazol-1-yloxy-tris(dimethylamino)-phosphonium hexafluorophosphate (BOP), benzotriazol-1-yloxy-tripyrrolidinophosphonium hexafluorophosphate (PyBOP), 7-aza-benzotriazol-1-yloxy-tripyrrolidino-phosphonium hexafluorophosphate (PyAOP), ethylcyano(hydroxyimino)acetato-O2 )-tri-(1-pyrrolidinyl)-phosphonium hexafluorophosphate (PyOxim), bromo-tripyrrolidino-phosphonium hexafluorophosphate (PyBrOP), 3-(diethoxy-phosphoryloxy)-1,2,3-benzo[d]triazin-4(3H)-one (DEPBT), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), 2-(1H-benzothiazolyl)-phosphonium hexafluorophosphate (PyOxim), bromo-tripyrrolidino-phosphonium hexafluorophosphate (PyBrOP), 3-(diethoxy-phosphoryloxy)-1,2,3-benzo[d]triazin-4(3H)-one (DEPBT), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), 2-(6-chloro-1H-benzotriazol-1-yl)-N,N,N',N'-tetramethylaminium hexafluorophosphate (HCTU), 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT), N-[(5-chloro- N,N',N'-methylmethanaminium hexafluorophosphate (HDMC), 1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (COMU), dimethylamino(triazolo[4,5-b]pyridin-3-yloxy)methylidene]-dimethylazanium tetrafluoroborate (TATU),N'-Tetramethyl-S-(1-oxido-2-pyridyl)thiouronium tetrafluoroborate (TOTT), tetramethylfluoroformamidinium hexafluorophosphate (TFFH), N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), 1-propanephosphonic anhydride (T3P), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM), dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide Coupling agents include, but are not limited to, (DIC), (N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDAC), or 1,1'-carbonyldimidizole (CDI). In some embodiments, the coupling agent in step (d) of method 2 can be HATU, TBTU, HBTU, CDMT, HDMC, COMU, TATU, TOTT, T3P, or DMTMM. In some embodiments, the coupling agent in step (d) of method 2 is (PyBOP).

[0168] It will be appreciated that step (d) of method 2 can be carried out in the presence of an additive. Suitable additives include, but are not limited to, 1-hydroxybenzotriazole (HOBt), 1-hydroxybenzotriazole-6-sulfonamidomethyl resin HCl (HOBt-6-sulfonamidomethyl resin HCl), hydroxy-3,4-dihydro-4-oxo-1,2,3-benzotriazine (HOOBt), N-hydroxysuccinimide (NHS), 1-hydroxy-7-aza-1H-benzotriazole (HOAt), ethyl 2-cyano-2-(hydroximino)acetate, or 4-(N,N-dimethylamino)pyridine (DMAP). It will be appreciated that step (d) of method 2 can be carried out at any temperature commonly used in solid phase synthesis, such as room temperature, under cooling, or under warm conditions. The coupling step (d) of Method 2 can be carried out for a period of time such as about 5 minutes to about 200 minutes, or about 10 minutes to about 100 minutes, or about 40 minutes to about 60 minutes to about 15 minutes, or about 30 minutes, or about 90 minutes, etc. Following the coupling step (d) of Method 2, the resin-based compound of Formula 7 prepared in step (d) of Method 2 can be washed with a reagent commonly known in the art such as DMF, IPA, CH2Cl2, etc.

[0169] In step (e) of method 2, PG on the resin-based compound of formula 7 4can be an amine protecting group such as 9-fluorenylmethyl-carbonyl (Fmoc), benzyl carbamate (Cbz), t-butyloxycarbonyl (Boc), acetamide, trifluoroacetamide, p-toluenesulfonamide, triphemylmethyl, monomethoxytrityl (MMt), and 4-methyltrityl (Mtt). The third deprotecting agent in step (e) of method 2 can be a reagent or mixture of reagents capable of removing a protecting group selected from the group consisting of 9-fluorenylmethyl-carbonyl (Fmoc), benzyl carbamate (Cbz), t-butyloxycarbonyl (Boc), acetamide, trifluoroacetamide, p-toluenesulfonamide, triphemylmethyl, monomethoxytrityl (MMt), and 4-methyltrityl (Mtt). In some embodiments, PG 4 can be 9-fluorenylmethyl-carbonyl (Fmoc). In some embodiments, the third deprotecting agent can be the third deprotecting agent of step (e) of method 2, and includes a reagent or mixture of reagents capable of removing 9-fluorenylmethyl-carbonyl. In some embodiments, the third deprotecting agent can be an acidic or basic deprotecting agent. In some embodiments, the third deprotecting agent can be a basic deprotecting agent, such as piperidine, morpholine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), diethylamine (DEA), dicyclohexamine, 4-methylpiperidine (4MP), tris(2-aminoethyl)amine, pyridine, or collidine. In some embodiments, the third deprotecting agent can be an acidic deprotecting agent, such as TFA, HCl, or H3PO4. Suitable deprotecting agents and conditions for step (e) of method 2 can be found, for example, in Wuts, PGM, Greene, TW, Greene, TW, & John Wiley & Sons. (2006). Greene's protective groups in organic synthesis. Hoboken, NJ: Wiley-Interscience.

[0170] In some embodiments, step (e) of method 2 can be carried out in a solvent such as a polar aprotic solvent. The polar aprotic solvent of step (e) of method 2 can be any polar aprotic solvent known in the art. Suitable polar aprotic solvents include, but are not limited to, dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO). It will be appreciated that step (e) of method 2 can be carried out at any temperature commonly used in solid phase synthesis, such as at room temperature, under cooling, or under warm conditions. The deprotection step (e) of method 2 can be carried out for a period of time such as about 5 minutes to about 200 minutes, or about 10 minutes to about 100 minutes, or about 40 minutes to about 60 minutes to about 45 minutes, or about 60 minutes, or about 90 minutes. Following the deprotection step (e) of Method 2, the resin-based compound of Formula 9 prepared in step (e) of Method 2 can be washed with reagents commonly known in the art, such as DMF, IPA, CH2Cl2, and the like.

[0171] In step (f) of method 2, PG 5 can be a carboxy protecting group. In some embodiments, PG 5 can be t-butyl. PG 5 For the selection of, see step (b) of method 2 above. In step (f) of method 2, the solvent can be a polar aprotic solvent. In some embodiments, the polar aprotic solvent can be any polar aprotic solvent commonly known in the art. Suitable polar aprotic solvents include, but are not limited to, dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO). In some embodiments, the polar aprotic solvent is DMF. Suitable bases for use in step (f) include N,N-diisopropylethylamine (i In some embodiments, the base in step (f) of method 2 is N,N-diisopropylethylamine (N,N-diisopropylethylamine), but is not limited to N,N-diisopropylethylamine (N,N-diisopropylethylamine). iThe coupling agent in step (f) of method 2 can be any coupling agent commonly known in the art for resin-based synthesis. Suitable coupling agents for use in step (f) of method 2 include benzotriazol-1-yloxy-tris(dimethylamino)-phosphonium hexafluorophosphate (BOP), benzotriazol-1-yloxy-tripyrrolidinophosphonium hexafluorophosphate (PyBOP), 7-aza-benzotriazol-1-yloxy-tripyrrolidino-phosphonium hexafluorophosphate (PyAOP), ethylcyano(hydroxyimino)acetato-O2 )-tri-(1-pyrrolidinyl)-phosphonium hexafluorophosphate (PyOxim), bromo-tripyrrolidino-phosphonium hexafluorophosphate (PyBrOP), 3-(diethoxy-phosphoryloxy)-1,2,3-benzo[d]triazin-4(3H)-one (DEPBT), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), 2-(1H-benzothiazolyl)-phosphonium hexafluorophosphate (PyOxim), bromo-tripyrrolidino-phosphonium hexafluorophosphate (PyBrOP), 3-(diethoxy-phosphoryloxy)-1,2,3-benzo[d]triazin-4(3H)-one (DEPBT), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), 2-(6-chloro-1H-benzotriazol-1-yl)-N,N,N',N'-tetramethylaminium hexafluorophosphate (HCTU), 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT), N-[(5-chloro- N,N',N'-methylmethanaminium hexafluorophosphate (HDMC), 1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (COMU), dimethylamino(triazolo[4,5-b]pyridin-3-yloxy)methylidene]-dimethylazanium tetrafluoroborate (TATU),N'-Tetramethyl-S-(1-oxido-2-pyridyl)thiouronium tetrafluoroborate (TOTT), tetramethylfluoroformamidinium hexafluorophosphate (TFFH), N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), 1-propanephosphonic anhydride (T3P), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM), dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide Coupling agents include, but are not limited to, (DIC), (N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDAC), and 1,1'-carbonyldimidizole (CDI). In some embodiments, the coupling agent in step (f) of method 2 can be HATU, TBTU, HBTU, CDMT, HDMC, COMU, TATU, TOTT, T3P, or DMTMM. In some embodiments, the coupling agent in step (f) of method 2 is (PyBOP).

[0172] It will be appreciated that step (f) of method 2 can be carried out in the presence of an additive. Suitable additives include, but are not limited to, 1-hydroxybenzotriazole (HOBt), 1-hydroxybenzotriazole-6-sulfonamidomethyl resin HCl (HOBt-6-sulfonamidomethyl resin HCl), hydroxy-3,4-dihydro-4-oxo-1,2,3-benzotriazine (HOOBt), N-hydroxysuccinimide (NHS), 1-hydroxy-7-aza-1H-benzotriazole (HOAt), ethyl 2-cyano-2-(hydroximino)acetate, or 4-(N,N-dimethylamino)pyridine (DMAP). It will be appreciated that step (f) of method 2 can be carried out at any temperature commonly used in solid phase synthesis, such as room temperature, under cooling, or under warm conditions. The coupling step (f) of Method 2 can be carried out for a period of time such as about 5 minutes to about 200 minutes, or about 10 minutes to about 100 minutes, or about 40 minutes to about 60 minutes to about 15 minutes, or about 30 minutes, or about 90 minutes, etc. Following the coupling step (f) of Method 2, the resin-based compound of Formula 10 prepared in step (f) of Method 2 can be washed with a reagent commonly known in the art such as DMF, IPA, CH2Cl2, etc.

[0173] In step (g) of method 2, PG on the resin-based compound of formula 10 2 , P.G. 3 and / or PG 5 may each be a protecting group, such as a carboxyl protecting group. In some embodiments, the third deprotecting agent in step (g) of method 2 may be a reagent or mixture of reagents capable of removing a protecting group selected from the group consisting of t-butyl, benzyl, 2-chlorotrityl (2Cl-Trt), 2,4-dimethoxybenzyl (Dmb), allyl, 1,1-dimethylallyl (Dma), and p-nitrobenzyl (pNB). In some embodiments, PG 2 , P.G. 3 , and P.G. 5 Each of is a t-butyl group. The third deprotecting agent in step (g) of method 2 is PG 2 , P.G. 3 , and / or P.G.5 Depending on the identity of the third deprotecting agent, it may be an acidic or basic deprotecting agent. In some embodiments, the third deprotecting agent may be an acidic deprotecting agent, such as TFA, HCl, or H3PO4. In some embodiments, the third deprotecting agent may be selected from the group consisting of trifluoroacetic acid (TFA), hydrofluoric acid (HF), triflic acid (TfOH), trifluoroacetic acid (TFA) / water / triisopropylsilane mixture, trifluoroacetic acid (TFA) / dichloromethane (DCM) mixture, triethylsaline (TES), indole, phenol / anisole mixture, and thioanisole. Suitable deprotecting agents and conditions for step (g) can be found, for example, in Wuts, PGM, Greene, TW, Greene, TW, & John Wiley & Sons. (2006). Greene's protective groups in organic synthesis. Hoboken, NJ: Wiley-Interscience. In some embodiments, the third deprotecting agent of step (g) of method 2 is a trifluoroacetic acid (TFA) / water / triisopropylsilane mixture. In some embodiments, the cleavage reagent of step (g) of method 2 is a trifluoroacetic acid (TFA) / water / triisopropylsilane mixture.

[0174] It will be appreciated that the organic solvent in step (g) of method 2 can be any organic solvent known in the art. Suitable organic solvents include, but are not limited to, CH2Cl2, THF, diethyl ether, MeOH, EtOH, acetone, acetonitrile, and the like. It will be appreciated that step (g) of method 2 can be carried out at any temperature commonly used in solid phase synthesis, such as at room temperature, under cooling, or under warm conditions. The deprotection step (g) of method 2 can be carried out for a period of time, such as about 5 minutes to about 200 minutes, or about 10 minutes to about 100 minutes, or about 40 minutes to about 60 minutes to about 45 minutes, or about 60 minutes, or about 90 minutes.

[0175] Method 2, specific embodiment: PSMA 617(I) can be prepared using solid phase peptide synthesis techniques, as shown in the following scheme. [ka] [ka]

[0176] The following reagents and intermediates can be used: [ka]

[0177] Three chemical transformations are performed. The Fmoc protecting group is removed from the Fmoc-Lys(Mtt)-Wang resin (2-1) by mixing the resin with a base in a polar aprotic solvent, and then the resin is washed with the solvent. In an organic solvent, L-glutamic acid di-tert-butyl ester hydrochloride (3') is converted to the activated urea or carbamate (2-3), for example, using a base and carbonyldiimidazole, p-nitrophenyl chloroformate, phenyl chloroformate, etc. The deprotected compound 2-2 is mixed with a solution of 2-3. Once the transformation to 2-4 is complete, the resin is washed with an organic solvent.

[0178] The methyltrityl protecting group is removed from 2-4 by mixing the resin with a mixture of 2,2,2-trifluoroethanol, acetic acid and dichloromethane. After removal of the trityl group, the resin is washed with a polar aprotic solvent. The deprotected compound 2-5 is mixed with (2S)-2-(9-fluorenylmethyloxycarbonylamino)-3-naphthalen-2-yl-propionic acid (2-6), a base, and an activating agent such as, but not limited to, a phosphonium salt activating agent in a polar aprotic solvent. After conversion to 2-7, the resin is washed with a polar aprotic solvent.

[0179] The Fmoc protecting group is removed from 2-7 by mixing the resin with a base in a polar aprotic solvent and washing the resin with the solvent. The deprotected compound 2-8 is mixed with 2-9, a base, and an activating agent, such as but not limited to a phosphonium salt activating agent, in a polar aprotic solvent. After conversion to 2-10, the resin is washed with a polar aprotic solvent and an organic solvent.

[0180] The tert-butyl groups are removed and the resin is cleaved by mixing with a mixture of trifluoroacetic acid, triisopropylsilane, and water. The liquid from the cleaved resin is separated, added to an organic solvent, and filtered. The crude PSMA 617 is purified by column chromatography and the material is desalted using a C18 column and an acetonitrile / water gradient eluent. The acetonitrile is evaporated or distilled from the aqueous solution under vacuum, and PSMA 617(I) is isolated from the solution by lyophilization. EXAMPLES

[0181] The examples and preparations provided below further describe and illustrate certain aspects of the embodiments of the present disclosure. It should be understood that the scope of the present disclosure is in no way limited by the scope of the following examples.

[0182] Abbreviation The examples described herein use materials that are known to those of skill in the art, including, but not limited to, those described by the following abbreviations:

[0183] [Table 1-1]

[0184] Example 1a: Preparation of AB2 Compound AB2 was prepared according to the following synthetic scheme: [ka] 403 mg of 4-nitrophenyl chloroformate (2.0 mmol, 0.95 equiv.) was added to a stirred solution of 621 mg of L-glutamic acid di-tert-butyl ester hydrochloride (2.1 mmol, 1.0 equiv.) in 21 mL of dichloromethane (CHCl) in a round-bottom flask under argon. The solution was cooled to 0° C., and then 0.730 mL of diisopropylethylamine ( i A solution of PrNEt) (4.2 mmol, 2.0 equiv) was added dropwise using a pressure equalizing addition funnel. i After the addition of PrNEt was complete, the reaction mixture was stirred for 30 min, warmed to room temperature, and stirred for an additional 30 min. The reaction mixture was then evaporated to give the crude product as an oily residue, which was used without further purification or kept under high vacuum until desired.

[0185] Example 1b: Alternative preparation of AB2 A reaction vessel was charged with 0.891 g of 4-nitrophenyl chloroformate (4.42 mmol) and 15 mL of dichloromethane (CHCl) and purged with argon. The reaction vessel was fitted with a temperature probe and the solution was cooled to below -10°C using an IPA / ice bath. In a separate flask, 1.44 g of L-glutamic acid di-tert-butyl ester hydrochloride (4.87 mmol) was dissolved in 15 mL of dichloromethane (CHCl), followed by 1.54 mL of diisopropylethylamine ( i L-Glutamic acid di-tert-butyl ester hydrochloride solution was added dropwise to the chilled 4-nitrophenyl chloroformate solution, maintaining the temperature at -11°C to -5°C. After the addition was complete, the ice bath was removed and the reaction mixture was stirred at room temperature for 4 hours, at which time the reaction was shown to be complete using UPLC / MS. The reaction mixture was used without further purification.

[0186] Example 2: Preparation of PSMA-617 The compound of formula I was prepared by SPPS starting from Fmoc-Lys(N-4-methoxytrityl)-2-chlorotrityl-resin. The compound of formula I was prepared according to the following synthesis scheme: [ka] [ka]

[0187] Tables 1 and 2 below summarize the above synthesis schemes, which were carried out according to the methods described herein:

[0188] [Table 1-2]

[0189] [Table 2]

[0190] Pretreatment process: Prior to use in the synthesis process, a peptide synthesis vessel was charged with 2.00 g (1.08 mmol) of Fmoc-Lys(Mtt)-Wang resin (resin loading = 0.54 mmol / g). The resin was washed with 20 mL of dimethylformamide (DMF) for 5 min and then drained from the vessel. The washing step was repeated one additional time.

[0191] Step 1: Urea formation The lysine resin was added to a peptide synthesis vessel. A solution of 20% piperidine in DMF (approximately 20 mL) for Fmoc deprotection was added. Argon was bubbled through the solution for 15 min and then vented. 20% piperidine in DMF (approximately 20 mL) was added and aeration was continued for 5 min and then vented (repeated twice). The resin was washed with DMF (approximately 20 mL x 3), followed by IPA (approximately 20 mL x 3) and finally with CH2Cl2 (approximately 20 mL x 3). The crude residue of AB2 was dissolved in 60 mL of CH2Cl2 and half the volume was added to the deprotected lysine resin followed by i PrNEt was added. Argon was bubbled through the solution for 45 min and then vented. The remaining crude AB2 solution was then i PrNEt was added to the resin and argon was bubbled through the solution for an additional 30 min. The reaction vessel was then evacuated and the resin was washed with CHCl (about 20 mL x 3), followed by IPA (about 20 mL x 3) and again with CHCl (about 20 mL x 3).

[0192] Alternative urea formation: The lysin resin was added to the peptide synthesis vessel. The AB2 reaction mixture from Example 1b was added to 0.188 mL of i The resin was added with PrNEt (1.08 mmol). The reaction mixture was stirred overnight (about 18 h). The resin was washed with CHCl (about 20 mL x 3).

[0193] Step 2: Mtt deprotection 2% TFA in CH2Cl2 (approximately 20 mL) was added and argon was bubbled through the solution for 5 min. Small amounts of CH2Cl2 were added to the reaction vessel to maintain the same volume in case of vigorous aeration. The yellow solution was then drained and repeated 5 times. The resin was washed with fresh CH2Cl2 until the filtrate remained clear. A 2% solution of TFA in CH2Cl2 was added once more. If the solution remained clear, the reaction mixture was drained and the next coupling step was carried out. If the solution turned yellow, the resin was washed with fresh CH2Cl2 until clear and this process was repeated until a clear reaction solution was obtained. The resin was washed with DMF (approximately 20 mL x 3).

[0194] Alternative Mtt deprotection A solution of 112 mL of CHCl, 32 mL of 2,2,2-trifluoroethanol (TFE), and 16 mL of acetic acid (AcOH) was prepared. 20 mL of the AcOH-TFE solution was added to the resin in the peptide synthesis vessel. The mixture was stirred for approximately 1 hour, and the liquid was drained from the peptide synthesis vessel. This procedure was repeated one more time. 20 mL of the AcOH-TFE solution was added to the resin in the peptide synthesis vessel, and the mixture was stirred overnight. The liquid was drained from the peptide synthesis vessel. 20 mL of the AcOH-TFE solution was added to the resin in the peptide synthesis vessel, and the mixture was stirred for approximately 1 hour. The liquid was drained from the peptide synthesis vessel. This procedure was repeated one more time. The resin was washed with 3×CHCl, 3×2-propanol (IPA), and 3×DMF.

[0195] Step 3: Amino coupling A solution of amino acid in DMF (approximately 20 mL), i Pr2NEt and PyBOP were added to a peptide synthesis vessel. Argon was bubbled through the solution for 1 h and then vented. The resin was washed with DMF (approximately 20 mL x 3), followed by IPA (approximately 20 mL x 3) and again with DMF (approximately 20 mL x 3).

[0196] Alternative Amino Coupling A solution of amino acid in DMF (approximately 20 mL), PyBOP in DMF (approximately 20 mL), and i Pr2NEt was added to the peptide synthesis vessel containing the resin. The mixture was stirred for about 4 hours, and then the liquid was drained from the peptide synthesis vessel. The resin was washed with DMF (about 20 mL x 3), followed by IPA (about 20 mL x 3).

[0197] Step 4: Fmoc deprotection A deprotection step was performed before each amino acid coupling step (in addition to Mtt deprotection using 2% TFA in CH2Cl2). For Fmoc deprotection, a solution of 20% piperidine in DMF (about 20 mL) was added. Argon was bubbled through the solution for 15 min and then evacuated. 20% piperidine in DMF (about 20 mL) was added and aeration was continued for 5 min before evacuating (2x). The resin was washed with DMF (about 20 mL x 3), followed by IPA (about 20 mL x 3) and again with DMF (about 20 mL x 3).

[0198] Alternative FMOC deprotection An alternative deprotection step was performed before each amino acid coupling step (in addition to Mtt deprotection). To the resin was added a solution of 20% piperidine in DMF (about 20 mL). The reaction was stirred for about 10 minutes. The liquid was drained from the reaction vessel and this process was repeated two more times. The resin was washed with DMF (about 20 mL x 3), followed by IPA (about 20 mL x 3). In some cases, the resin was further washed with DMF (about 20 mL x 3) or alternatively with CH2Cl2 (about 20 mL x 3).

[0199] Process 5: Resin cutting The resin was washed with CH2Cl2 (approximately 20 mL x 3) and drained. 25 mL of cleavage reagent (95% TFA, 2.5% H20, 2.5% triisopropylsilane) was added to the peptide synthesis vessel, bubbled with argon for 1 h, evacuated, and repeated twice with fresh cleavage reagent (10 mL, 15 min). The filtrate was stirred overnight at room temperature under argon. The reaction mixture was concentrated under reduced pressure until 10 mL remained. The product was triturated in 40 mL of diethyl ether and centrifuged. The solution was decanted from the resulting pellet. The previous step was repeated twice by resuspending the pellet in 50 mL of diethyl ether and centrifuging. The pellet was dried over a stream of argon and then dried under high vacuum.

[0200] Alternative Resin Cutting A mixture of 42 mL of trifluoroacetic acid (TFA), 0.90 mL of water, and 3.0 mL of triisopropylsilane (TIPS) was prepared and added to the resin in a peptide synthesis vessel. The mixture was stirred for 6 hours (the last hour was warmed to 30° C. in a water bath). The resin was filtered and the solid was washed with 2 mL of TFA. The product containing the filtrate and washings was combined and washed with 420 mL of methyl-t-butyl ether (MTBE) cooled in an ice-water bath. The solid product was collected by filtration. The solid was washed with 5 mL of MTBE. The solid was dried under vacuum to provide 1.19 g of crude compound of formula I.

[0201] Purification step 1 The resulting powder was dissolved in a small amount of DMSO and loaded onto a C18 column. The desired product was purified by reverse phase chromatography (0-55% acetonitrile in 50 mM ammonium carbonate buffer, pH 7.0). Acetonitrile was evaporated under reduced pressure and the remaining aqueous buffer solution was frozen and removed by lyophilization. 256 mg (49.2%) of the desired product was recovered as a white powder.

[0202] Purification step 2 The compound of formula I was purified using a Biotage 60g C18 ULTRA column with mobile phase A = 0.1% TFA in water and mobile phase B = acetonitrile (ACN) (flow rate = 45 mL / min, 1 CV = 90 mL, collection wavelength at 275 nm). Crude PSMA-617 in 20 mL of 0.1% TFA in water with 3 mL of ACN is loaded onto the column. Purification was performed with 1 CV 15% B, 7 CV 15% B to 25% B, held at 25% B. The product peak began to elute at approximately 5 CV and fractions were collected. Fractions were examined using UPLC-MS and acceptable product fractions (>= 98% at 275 nm) were combined and the product solution was concentrated using a rotary evaporator to remove ACN (final volume = 59 mL). The compound of formula I was desalted using a Biotage 30g C18 ULTRA column, which was set up with mobile phase A=0.1% AcOH in water and mobile phase B=ACN (flow rate=25mL / min, 1CV=45mL, collection wavelength at 275nm). To aqueous PSMA-617 (59mL) was added 0.28g ammonium acetate, 0.36mL AcOH, and 3mL ACN. The PSMA-617 solution was loaded onto the column and the column was washed with 5CV 5% B. The product was eluted from the column with 3CV 39% B. The product peak eluted in 6CV and 1 fraction was collected. The product solution was concentrated using a rotary evaporator to remove ACN. The solid was isolated by lyophilization to provide 0.675g of the compound of formula I.

[0203] Example 3: Characterization of PSMA-617 by Nuclear Magnetic Resonance and High Resolution MS ES+(TOF): 1H NMR (D6-DMSO): 8.03(t, 1H), 7.88(t, 1H), 7.84(d, 1H), 7.69-7.80(m, 2H), 7.69(s, 1H), 7.43-7.47(m, 2 H), 7.39(dd, 1H), 6.29-6.33(m, 2H), 4.50-4.52(m, 1H), 4.08-4.11(m, 1H), 3.99-4.02(m, 1H), 3.46(br s, 6H), 3.13(br s, 2H), 3.10(d, 1H), 2.89-3.04(m, 17H), 2.68(s, 4H), 2.22-2.27(m, 2H), 2.07(s, 1H), 1.91(s, 1H), 1.57~1 .74(m,5H),1.46~1.50(m,2H),1.31~1.35(m,3H),1.22~1.26(m,3H),1.06~1.12(m,1H),0.80~0.82(m,2H).

[0204] NOTE: Six carboxylic acid protons are spread across the spectrum, with additional resonances from residual acetic acid (1.91 ppm) and acetonitrile (2.07 ppm).

[0205] 13 C NMR(D6-DMSO):175.09, 174.52, 174.14, 173.73, 170.99, 170.55, 170.11, 169.64, 157 .26, 135.78, 132.86, 131.70, 127.81, 127.37, 127.32, 127.24, 127.22, 125.83, 125.24 , 57.80, 55.34, 54.86, 53.85, 52.33, 51.78, 51.24, 50.52, 49.96, 49.84, 44.54, 43.77, 38.14, 37.86, 36.81, 31.65, 30.05, 29.61, 29.50, 28.63, 28.56, 28.38, 27.69, 22.41.

[0206] Note: Additional resonances are present from residual acetic acid (171.88 ppm, 20.98 ppm) and acetonitrile (1.068 ppm).

[0207] High resolution MS ES+(TOF):M+H +=1042.5067m / z (theoretical value 1042.5096m / z) The present invention includes the following embodiments. [Claim 1] Formula I [ka] A process for preparing a compound of the formula: a) Formula A [ka] contacting the resin-based compound of formula A' with a first deprotecting agent in a polar aprotic solvent to produce a compound of formula A' [ka] or providing a resin-based compound of b) reacting a resin-based compound of formula A' with a compound of formula B [ka] in the presence of an organic solvent and a base to form a compound of formula C [ka] or providing a resin-based compound of c) contacting the resin-based compound of formula C with a second deprotecting agent in the presence of an organic solvent to produce a compound of formula C' [ka] or providing a resin-based compound of d) reacting a resin-based compound of formula C' with PG in the presence of a polar aprotic solvent, a coupling agent, and a base. 1 -3-(2-naphthyl)-L-alanine to form a compound of formula D [ka] or providing a resin-based compound of e) contacting the resin-based compound of formula D with a first deprotecting agent in a polar aprotic solvent to produce a resin-based compound of formula D' [ka] or providing a resin-based compound of f) reacting a resin-based compound of formula D' with PG in the presence of a polar aprotic solvent, a coupling agent and a base. 1 - contacting with tranexamic acid to form a compound of formula E

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Claims

1. Formula I 【Chemistry 1】 A process for preparing a compound of the formula: a) Formula A 【Chemistry 2】 contacting the resin-based compound of formula A' with a first deprotecting agent in a polar aprotic solvent to form a 【Chemistry 3】 To provide a resin-based compound according to the present invention; b) reacting a resin-based compound of formula A′ with a compound of formula B 【Chemistry 4】 in the presence of an organic solvent and a base to form a compound of formula C 【Chemistry 5】 To provide a resin-based compound according to the present invention; c) contacting the resin-based compound of formula C with a second deprotecting agent in the presence of an organic solvent to produce a compound of formula C' 【Chemistry 6】 To provide a resin-based compound according to the present invention; d) reacting a resin-based compound of formula C′ with PG in the presence of a polar aprotic solvent, a coupling agent, and a base. １ -3-(2-naphthyl)-L-alanine to form a compound of formula D 【Chemistry 7】 To provide a resin-based compound according to the present invention; e) contacting the resin-based compound of formula D with a first deprotecting agent in a polar aprotic solvent to produce a compound of formula D' 【Chemistry 8】 To provide a resin-based compound according to the present invention; f) reacting a resin-based compound of formula D' with PG in the presence of a polar aprotic solvent, a coupling agent and a base. １ - tranexamic acid to form a compound of formula E 【Chemistry 9】 To provide a resin-based compound according to the present invention; g) contacting the resin-based compound of formula E with a first deprotecting agent in a polar aprotic solvent to produce a compound of formula E' 【Chemistry 10】 To provide a resin-based compound according to the present invention; h) reacting a resin-based compound of formula E′ with a compound of formula E ２’ 【Chemistry 11】 in the presence of a polar aprotic solvent, a coupling agent, and a base to obtain a compound of formula F 【Chemistry 12】 To provide a resin-based compound according to the present invention; i) contacting said resin-based compound of formula F with a third deprotecting and / or cleaving agent in an organic solvent; In the formula, P.G. １ is 9-fluorenylmethyl-carbonyl (Fmoc), and PG ２ is 4-methyltrityl (Mtt), LG is 4-nitrophenoxy, R １ is C １ ~C ４ alkyl or cyclohexyl.

2. 2. The method of claim 1, wherein the first deprotecting agent in step (a) is selected from the group consisting of piperidine, morpholine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), diethylamine (DEA), dicyclohexamine, 4-methylpiperidine (4MP), tris(2-aminoethyl)amine, pyridine, and collidine.

3. 3. The method of claim 2, wherein the first deprotecting agent in step (a) is piperidine.

4. 4. The method of claim 3, wherein the polar aprotic solvent in step (a) is selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO).

5. 5. The method of claim 4, wherein the polar aprotic solvent in step (a) is dimethylformamide (DMF).

6. The base in step (b) is N,N-diisopropylethylamine ( ｉ P ２ 2. The method of claim 1, wherein the aryl group is selected from the group consisting of aryl ether (AEE), aryl ether (BEE), triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine.

7. The base in the step (b) is N,N-diisopropylethylamine ( ｉ P ２ 7. The method of claim 6, wherein said aryloxycarbonyl group is aryloxycarbonyl, ...

8. The organic solvent in step (b) is CH ２ C ２ 8. The method of claim 7, wherein the solvent is selected from the group consisting of THF, diethyl ether, MeOH, EtOH, acetone, and acetonitrile.

9. The organic solvent in step (b) is CH ２ C ２ The method according to claim 8, wherein

10. 2. The method of claim 1, wherein the second deprotecting agent in step (c) is selected from the group consisting of trifluoroacetic acid (TFA), acetic acid, trifluoroethanol, dichloromethane, hexafluoroisopropanol, dichloromethane, and combinations thereof.

11. 11. The method of claim 10, wherein the second deprotecting agent in step (c) is trifluoroacetic acid (TFA) or a mixture of acetic acid and trifluoroethanol.

12. The organic solvent in step (c) is CH ２ C ２ 12. The method of claim 11, wherein the solvent is selected from the group consisting of THF, diethyl ether, MeOH, EtOH, acetone, and acetonitrile.

13. The organic solvent in step (c) is CH ２ C ２ The method of claim 12, wherein

14. 2. The method of claim 1, wherein the polar aprotic solvent in step (d) is selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO).

15. 15. The method of claim 14, wherein the polar aprotic solvent in step (d) is dimethylformamide (DMF).

16. The base in step (d) is N,N-diisopropylethylamine ( ｉ P ２ 16. The method of claim 15, wherein the aryl group is selected from the group consisting of: triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine.

17. The base in step (d) is N,N-diisopropylethylamine ( ｉ P ２ 17. The method of claim 16, wherein said aryloxycarbonyl group is aryloxycarbonyl, ...

18. The coupling agent in step (d) is selected from the group consisting of benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT), N-[(5-chloro-3-oxide-1H-benzotriazol-1-yl)-4-morpholinylmethylene]-N-methyl ...

18. The method of claim 17, wherein the dimethylamino group is selected from the group consisting of thanaminium hexafluorophosphate (HDMC), 1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (COMU), dimethylamino(triazolo[4,5-b]pyridin-3-yloxy)methylidene]-dimethylazanium; tetrafluoroborate (TATU), N,N,N',N'-tetramethyl-S-(1-oxido-2-pyridyl)thiouronium tetrafluoroborate (TOTT), N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), 1-propanephosphonic anhydride (T3P), and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM).

19. 19. The method of claim 18, wherein the coupling agent in step (d) is benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP).

20. 2. The method of claim 1, wherein the first deprotecting agent in step (e) is selected from the group consisting of piperidine, morpholine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), diethylamine (DEA), dicyclohexamine, 4-methylpiperidine (4MP), tris(2-aminoethyl)amine, pyridine, and collidine.

21. 21. The method of claim 20, wherein the first deprotecting agent in step (e) is piperidine.

22. 22. The method of claim 21, wherein the polar aprotic solvent in step (e) is selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO).

23. 23. The method of claim 22, wherein the polar aprotic solvent in step (e) is dimethylformamide (DMF).

24. 2. The method of claim 1, wherein the polar aprotic solvent in step (f) is selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO).

25. 25. The method of claim 24, wherein the polar aprotic solvent in step (f) is dimethylformamide (DMF).

26. The base in step (f) is N,N-diisopropylethylamine ( ｉ P ２ 26. The method of claim 25, wherein the aryl group is selected from the group consisting of: triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine.

27. The base in step (f) is N,N-diisopropylethylamine ( ｉ P ２ 27. The method of claim 26, wherein said aryloxycarbonyl group is aryloxycarbonyl, ...

28. The coupling agent in step (f) is selected from the group consisting of benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT), N-[(5-chloro-3-oxide-1H-benzotriazol-1-yl)-4-morpholinylmethylene]-N-methylmethanamine 28. The method of claim 27, wherein the dimethylamino-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride is selected from the group consisting of N,N,N',N'-tetramethyl-S-(1-oxido-2-pyridyl)thiouronium tetrafluoroborate (TATU), N,N,N',N'-tetramethyl-S-(1-oxido-2-pyridyl)thiouronium tetrafluoroborate (TOTT), N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), 1-propanephosphonic anhydride (T3P), and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM).

29. 29. The method of claim 28, wherein the coupling agent in step (f) is benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP).

30. 2. The method of claim 1, wherein the first deprotecting agent in step (g) is selected from the group consisting of piperidine, morpholine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), diethylamine (DEA), dicyclohexamine, 4-methylpiperidine (4MP), tris(2-aminoethyl)amine, pyridine, and collidine.

31. 31. The method of claim 30, wherein the first deprotecting agent in step (g) is piperidine.

32. 32. The method of claim 31 , wherein the polar aprotic solvent in step (g) is selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO).

33. 33. The method of claim 32, wherein the polar aprotic solvent in step (g) is dimethylformamide (DMF).

34. 2. The method of claim 1, wherein the polar aprotic solvent in step (h) is selected from the group consisting of dimethylformamide (DMF), N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), dichloromethane / dimethylformamide mixtures, acetonitrile (ACN), acetonitrile / dimethylformamide mixtures, and dimethylsulfoxide (DMSO).

35. 35. The method of claim 34, wherein the polar aprotic solvent in step (h) is dimethylformamide (DMF).

36. The base in step (h) is N,N-diisopropylethylamine ( ｉ P ２ 36. The method of claim 35, wherein the aryl group is selected from the group consisting of: triethylamine (TET), triethylamine (TEA), 4-methylmorpholine (NMM), imidazole, pyridine, and collidine.

37. The base in step (h) is N,N-diisopropylethylamine ( ｉ P ２ NEt).

38. The coupling agent in step (h) is selected from the group consisting of benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT), N-[(5-chloro-3-oxide-1H-benzotriazol-1-yl)-4-morpholinylmethylene]-N-methylmethanamine 38. The method of claim 37, wherein the dimethylamino-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride is selected from the group consisting of N,N,N',N'-tetramethyl-S-(1-oxido-2-pyridyl)thiouronium tetrafluoroborate (TATU), N,N,N',N'-tetramethyl-S-(1-oxido-2-pyridyl)thiouronium tetrafluoroborate (TOTT), N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), 1-propanephosphonic anhydride (T3P), and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM).

39. 39. The method of claim 38, wherein the coupling agent in step (h) is benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP).

40. 2. The method of claim 1, wherein the third deprotecting agent of step (i) is selected from the group consisting of trifluoroacetic acid (TFA), hydrofluoric acid (HF), triflic acid (TfOH), trifluoroacetic acid (TFA) / water / triisopropylsilane mixture, trifluoroacetic acid (TFA) / dichloromethane (DCM) mixture, triethylsaline (TES), indole, phenol / anisole mixture, and thioanisole.

41. 41. The method of claim 40, wherein the third deprotecting agent of step (i) is a trifluoroacetic acid (TFA) / water / triisopropylsilane mixture.

42. 42. The method of claim 41, wherein the cleavage reagent in step (i) is trifluoroacetic acid (TFA) or a trifluoroacetic acid (TFA) / water / triisopropylsilane mixture.

43. The organic solvent in step (i) is CH ２ C ２ 43. The method of claim 42, wherein the solvent is selected from the group consisting of THF, diethyl ether, MeOH, EtOH, acetone, and acetonitrile.

44. 44. The method of claim 43, wherein the cleavage reagent in step (i) is a trifluoroacetic acid (TFA) / water / triisopropylsilane mixture.

45. The organic solvent in step (i) is CH ２ C ２ 45. The method of claim 44, wherein:

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