Solution-phase peptide-supported synthesis of peptides and peptidomimetics

Abstract

The present invention provides compounds that are useful as supports for solution phase organic synthesis, e.g., for the solution phase organic synthesis of peptides and peptidomimetics. In one aspect, the present invention provides compounds that are useful as supports for solution phase organic synthesis, e.g., for the solution phase organic synthesis of peptides and peptidomimetics, 1 , R 2 , R 3 [0010] Provided are compounds having the structure of Formula (I), where m, and n are as described herein, as well as methods of making and using the same. [Formula 1] TIFF2026508282000564.tif6183

Description

[Technical Field]

[0001] The present disclosure relates to liquid phase peptide support synthesis of peptides and peptidomimetics, pharmaceutical compositions, and methods for treating GLP1R-associated diseases. [Background technology]

[0002] The difference between liquid-phase peptide synthesis (LPPS) and solid-phase peptide synthesis (SPPS) is that the latter is supported on a large solid polymer, whereas the former is supported on a well-defined molecule. The difference between LPPS and CSPS is the difference between isolated precipitation in LPPS and phase separation in CSPS. SPPS is the well-established, most common, and preferred method for preparing polypeptides (Jaradat D., "Thirteen Decades of Peptide Synthesis: Key Developments in Solid Phase Peptide Synthesis and Amide Bond Formation Utilized in Peptide Ligation," Amino Acids 50(1):39-68 (2018)). However, due to its heterogeneity, large molar excesses of amino acid building blocks are required to ensure complete peptide chain elongation. CSPS, on the other hand, requires fewer molar equivalents of building blocks, but isolation and purification are laborious and time-consuming. It is not easy to generate suitable chemistry related to LPPS to eliminate the need for purification steps and to reduce the molar equivalents of building blocks, especially unnatural amino acids, in the synthesis of peptidomimetics, which is problematic for scale-up and overall material consumption.

[0003] Initial approaches using the known LPPS tag (tag 1 in Figure 1) to prepare a 10-mer peptidomimetic (M1) composed of five natural and five unnatural amino acid residues (Takahashi et al., "Novel Diphenylmethyl-Derived Amide Protecting Group for Efficient Liquid-Phase Peptide Synthesis: AJIPHASE," Org Lett. 14(17):4514-4517 (2012); Takahashi et al., "Novel Diphenylmethyl-Derived Amide Protecting Group for Efficient Liquid-Phase Peptide Synthesis: AJIPHASE," Org Lett. 14(17):4514-4517 (2012)) were unsuccessful. Peptide chain elongation could only reach the seventh amino acid residue due to insufficient separation of the tag-bearing peptide (TCP) from the reaction mixture.

[0004] Thus, there is a need in the art for new tag carriers that have better separation of tag-bearing peptides from reaction mixtures. In certain embodiments, the present disclosure fulfills this need and provides other advantages.

[0005] The foregoing discussion is presented solely to provide a better understanding of the nature of the problems facing the art and should not be construed in any way as an admission of prior art, nor should the recitation of any reference herein be construed as an admission that such reference constitutes "prior art" to the present application. Summary of the Invention

[0006] Various non-limiting aspects and embodiments of the disclosure are described below.

[0007] In one aspect, the present disclosure provides a compound of formula (I):

[0008] [ka] [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R 1 is C 1~5 is alkyl, R 2 is, independently at each occurrence, optionally replaced by C 10~30 is alkyl, R 3 is, independently at each occurrence, optionally replaced by C 10~30 alkyl] The present invention provides a compound having the structure:

[0009] In some embodiments, R 1 is methyl.

[0010] In some embodiments, R 2 is, independently at each occurrence, optionally replaced by C 18~26 alkyl, and R 3 is, independently at each occurrence, optionally replaced by C 18~26 In some embodiments, each R 2 are independently optionally substituted C 22 alkyl, and each R 3 are independently optionally substituted C 22 It is alkyl.

[0011] In some embodiments, the compound has the formula (Ia):

[0012] [ka] or a salt thereof.

[0013] In another aspect, the present disclosure provides a compound of formula (I):

[0014] [ka] [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R 1 is C 1~5 is alkyl, R 2 is, independently at each occurrence, optionally replaced by C 10~30 is alkyl, R 3 is, independently at each occurrence, optionally replaced by C 10~30 alkyl] or a salt thereof, the process comprising: (a) Structure:

[0015] [ka] [In the formula, wherein PG is a suitable protecting group; and (b) forming a compound of formula (I) from a compound of formula (II); Includes:

[0016] In some embodiments, PG is selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

[0017] In some embodiments, the compound of Formula (II) has the structure of Formula (IIa):

[0018] [ka]

[0019] In some embodiments, step (b) of forming the compound of Formula (I) comprises reacting the compound of Formula (II) with a deprotecting agent to produce the compound of Formula (I). In some embodiments, the deprotecting agent is selected from the group consisting of Pd(PPh)3, PhSiH3, H2, piperidine, and trifluoroacetic acid (TFA).

[0020] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0021] [ka] [In the formula, PG 1 is a suitable protecting group, PG 2 is a suitable protecting group; and Before step (a), forming a compound of formula (II) from a compound of formula (III). Further includes:

[0022] In some embodiments, PG 1 are independently selected from the group consisting of tert-butyldimethylsilyl (TBS), trimethylsilyl (TMS), triethylsilyl (TES), tert-butyldiphenylsilyl (TBDPS), and triisopropylsilyl (TIPS).

[0023] In some embodiments, PG 2 are independently selected from the group consisting of tert-butyldimethylsilyl (TBS), trimethylsilyl (TMS), triethylsilyl (TES), tert-butyldiphenylsilyl (TBDPS), and triisopropylsilyl (TIPS).

[0024] In some embodiments, each PG 1 and each PG 2 are identical. In some embodiments, PG 1 and P.G.2 are different from each other.

[0025] In some embodiments, the compound of Formula (III) has the structure of Formula (IIIa):

[0026] [ka]

[0027] In some embodiments, the step of forming a compound of formula (II) prior to step (a) comprises: reacting a compound of formula (III) with a deprotecting agent to obtain a deprotected intermediate; The intermediate is reacted with the compound of formula (IV): R-LG(IV) [In the formula, R is an optionally substituted C 10~30 is alkyl, wherein LG is a suitable leaving group, Producing a compound of formula (II) Includes:

[0028] In some embodiments, LG is selected from the group consisting of halogen, OTf, OMs, and OTs.

[0029] In some embodiments, the deprotecting agent is selected from the group consisting of tetrabutylammonium fluoride (TBAF), NH 4 F, SiF 4 , AcOH, HCl, LiAlH 4 , and K 2 CO 3 .

[0030] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0031] [ka] providing a compound of formula (V) having the formula: forming a compound of formula (III) from a compound of formula (V); Further includes:

[0032] In some embodiments, the step of forming a compound of formula (III) comprises: Compounds of formula (V) can be reacted with compounds of formula (VIa) PG-X(VIa) or Formula (VIb) PG2O(VIb) [In the formula, X is Cl or Br, Producing a compound of formula (III) Includes:

[0033] In some embodiments, the compound of Formula (VIa) or Formula (VIb) is selected from the group consisting of AllocCl, Alloc2O, Cbz2O, CbzCl, FmocCl, and Boc2O.

[0034] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0035] [ka] [In the formula, PG 3 is a suitable protecting group; and forming a compound of formula (V) from a compound of formula (VII) Further includes:

[0036] In some embodiments, PG 3 is selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

[0037] In some embodiments, the compound of formula (VII) has the following structure:

[0038] [ka]

[0039] In some embodiments, the step of forming a compound of Formula (V) comprises reacting a compound of Formula (VII) with a deprotecting agent to produce a compound of Formula (V). In some embodiments, the deprotecting agent is selected from the group consisting of Pd(PPh)3, PhSiH3, H2, piperidine, and trifluoroacetic acid (TFA).

[0040] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0041] [ka] providing a compound of formula (VIII) having the formula forming a compound of formula (VII) from a compound of formula (VIII) Further includes:

[0042] In some embodiments, the process for forming a compound of Formula (VII) comprises reacting a compound of Formula (VIII) with the structure:

[0043] [ka] with a compound of formula (IX) having the formula:

[0044] In some embodiments, the compound of formula (VIII) has the following structure:

[0045] [ka]

[0046] In some embodiments, the compound of formula (IX) has the following structure:

[0047] [ka]

[0048] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0049] [ka] [In the formula, PG 4 is a suitable protecting group; and forming a compound of formula (VIII) from a compound of formula (X); Further includes:

[0050] In some embodiments, PG 4 is selected from the group consisting of allyl, allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

[0051] In some embodiments, the compound of Formula (X) has the following structure:

[0052] [ka]

[0053] In some embodiments, the step of forming a compound of Formula (VIII) comprises reacting a compound of Formula (X) with a deprotecting agent, in some embodiments, the deprotecting agent is selected from the group consisting of Pd / K2CO3, 1,3-dimethylbarbituric acid / Pd(PPh3)4, Pd(PPh)3, PhSiH3, H2, piperidine, and trifluoroacetic acid (TFA).

[0054] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0055] [ka] providing a compound of formula (XI) having the formula forming a compound of formula (X) from a compound of formula (XI) Further includes:

[0056] In one embodiment, the compound of formula (XI) has the structure:

[0057] [ka]

[0058] In some embodiments, the process for forming a compound of formula (X) comprises: The compound of formula (XI) may be reacted with a compound of formula (XII):

[0059] [ka] or a salt thereof, Producing a compound of formula (X) Includes:

[0060] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0061] [ka] providing a compound of formula (XIII) having the formula forming a compound of formula (XI) from a compound of formula (XIII) Further includes:

[0062] In some embodiments, the compound of formula (XIII) has the following structure:

[0063] [ka]

[0064] In some embodiments, the process for forming a compound of Formula (XI) comprises reacting a compound of Formula (XIII) with an oxidizing agent to produce a compound of Formula (XI). In some embodiments, the oxidizing agent may be selected from the group consisting of pyridinium chlorochromate (PCC), pyridinium dichromate (PDC), silver carbonate (AgCO), tetra-n-propylammonium perruthenate (TPAP), and Dess-Martin periodinane (DMP).

[0065] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0066] [ka] [In the formula, Y is -OC 1~6 wherein R is an alkyl group; and forming a compound of formula (XIII) from a compound of formula (XIV); Further includes:

[0067] In one embodiment, the compound of formula (XIV) has the structure:

[0068] [ka]

[0069] In some embodiments, the process for forming the compound of Formula (XIII) comprises reacting the compound of Formula (XIV) with a reducing agent, in some embodiments, the reducing agent is selected from the group consisting of LiAlH, DIBALH, and LiBH.

[0070] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0071] [ka] providing a compound of formula (XV) having the formula forming a compound of formula (XIV) from a compound of formula (XV) Further includes:

[0072] In some embodiments, the compound of formula (XV) has the structure:

[0073] [ka]

[0074] In some embodiments, the process for forming a compound of formula (XIV) comprises reacting a compound of formula (XV) with a protecting group introducing agent to produce a compound of formula (XIV). In some embodiments, the protecting group introducing agent is selected from the group consisting of TBSCl, TBSOTf, TMSCl, TMSOTf, TESCl, TESOTf, TBDPSCl, TBDPSOTf, TIPSCl, and TIPSOTf.

[0075] In yet another aspect, the present disclosure provides a product prepared according to any of the methods described in this disclosure.

[0076] In some embodiments, the product has formula (I):

[0077] [ka] [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R 1 is C 1~5 is alkyl, R 2 is, independently at each occurrence, optionally replaced by C 10~30 is alkyl, R 3is, independently at each occurrence, optionally replaced by C 10~30 alkyl] or a salt thereof.

[0078] In one embodiment, the product has formula (Ia):

[0079] [ka] or a salt thereof.

[0080] In a further aspect, the present disclosure provides a compound of formula (I):

[0081] [ka] [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R 1 is C 1~5 is alkyl, R 2 is, independently at each occurrence, optionally replaced by C 10~30 is alkyl, R 3 is, independently at each occurrence, optionally replaced by C 10~30 alkyl] or a salt thereof, Methods for making peptides or peptidomimetics are provided.

[0082] In some embodiments, the compounds of formula (I) are used as supports or tags for generating peptides or peptidomimetics.

[0083] In one embodiment, the peptidomimetic has the formula:

[0084] [ka] or a salt thereof.

[0085] In one embodiment, the peptidomimetic has the formula:

[0086] [ka] or a salt thereof.

[0087] In another aspect, the present disclosure provides a compound of formula (I):

[0088] [ka] [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R 1 is C 1~5 is alkyl, R 2 is optionally replaced by C 10~30 independently selected from alkyl, R 3 is optionally replaced by C for each occurrence. 10~30 alkyl] or a salt thereof.

[0089] In one embodiment, the compound has the formula:

[0090] [ka] or a salt thereof.

[0091] In one embodiment, the compound has the formula:

[0092] [ka] or a salt thereof.

[0093] In yet another aspect, provided herein is a pharmaceutical composition comprising a peptidomimetic or compound of any of the embodiments described herein.

[0094] In a further aspect, provided herein is a pharmaceutical dosage form comprising a peptidomimetic or compound of any of the embodiments described herein.

[0095] In another aspect, the present invention provides a method for selectively targeting GLP1R on the surface of a cell using any compound or peptidomimetic of any of the embodiments described herein.In some embodiments, the cell is a mammalian cell.In some embodiments, the cell is a human cell.

[0096] In another aspect, provided herein is a method of enhancing activity of GLP1R in an individual in need thereof, comprising administering to the individual an effective amount of a peptidomimetic or compound of any of the embodiments described herein, a composition described herein, or a dosage form described herein.

[0097] In yet another aspect, provided herein is a method of lowering blood glucose levels in an individual in need thereof, comprising administering to the individual an effective amount of a peptidomimetic or compound of any of the embodiments described herein, a composition described herein, or a dosage form described herein.

[0098] In a further aspect, provided herein is a method of reducing body weight in an individual in need thereof, comprising administering to the individual an effective amount of a peptidomimetic or compound of any of the embodiments described herein, a composition described herein, or a dosage form described herein.

[0099] In another aspect, provided herein is a method for treating a GLP1R-related disease in an individual in need thereof, comprising administering to the individual an effective amount of any of the peptidomimetics or compounds of the embodiments described herein, the compositions described herein, or the dosage forms described herein. In some embodiments, the GLP1R-related disease is type II diabetes, obesity, liver disease, coronary artery disease, or kidney disease. In some embodiments, the GLP1R-related disease is type II diabetes and / or obesity. In some embodiments, the peptidomimetics or compounds of the embodiments described herein, the compositions described herein, or the dosage forms described herein are administered subcutaneously, intravenously, intradermally, intraperitoneally, or intramuscularly.

[0100] In yet another aspect, there is provided a method of making a peptide or peptidomimetic, comprising: (a) Formula (I):

[0101] [ka] [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R 1 is C 1~5 is alkyl, R 2 is optionally replaced by C for each occurrence. 10~30 independently selected from alkyl, R 3 is optionally replaced by C for each occurrence. 10~30 alkyl] or a salt thereof, (b) providing a first amino acid (AA1), and (c) coupling a first amino acid (AA1) with the compound of formula (I) to form a peptide bond between the first amino acid (AA1) and the compound of formula (I); Provided herein is a method comprising:

[0102] In some embodiments, the first amino acid (AA1) contains a protected amino group.

[0103] In some embodiments, the first amino acid (AA1) is:

[0104] [ka]

[0105] In some embodiments, the method of making a peptide or peptidomimetic may include activating a chemical group on the first amino acid (AA1) to prepare the first amino acid (AA1) for coupling with a compound of formula (I) prior to step (c).

[0106] In some embodiments, the method of making a peptide or peptidomimetic comprises: (d) removing the protecting group from the first amino acid (AA1); (e) providing a second amino acid (AA2), and (f) coupling a second amino acid (AA2) with the first amino acid (I-AA1) to form a peptide bond (I-AA1-AA2) between the second amino acid (AA2) and the first amino acid; may include:

[0107] In some embodiments, the second amino acid (AA2) is:

[0108] [ka]

[0109] In some embodiments, the method of making a peptide or peptidomimetic comprises: (g) removing the protecting group from the second amino acid (AA2); (h) providing a third amino acid (AA3), and (i) coupling a third amino acid (AA3) with a second amino acid to form a peptide bond (I-AA1-AA2-AA3) between the third amino acid (AA3) and the second amino acid; may include:

[0110] In some embodiments, the third amino acid (AA3) is:

[0111] [ka]

[0112] In some embodiments, the method of making a peptide or peptidomimetic comprises: (j) removing the protecting group from the third amino acid (AA3); (k) providing a fourth amino acid (AA4), and (l) coupling a fourth amino acid (AA4) with a third amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4) between the fourth amino acid (AA4) and the third amino acid; may include:

[0113] In some embodiments, the fourth amino acid (AA4) is:

[0114] [ka]

[0115] In some embodiments, the method of making a peptide or peptidomimetic comprises: (m) removing the protecting group from the fourth amino acid (AA4); (n) providing a fifth amino acid (AA5), and (o) coupling a fifth amino acid (AA5) with a fourth amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5) between the fifth amino acid (AA5) and the fourth amino acid; may include:

[0116] In some embodiments, the fifth amino acid (AA5) is:

[0117] [ka]

[0118] In some embodiments, the method of making a peptide or peptidomimetic comprises: (p) removing the protecting group from the fifth amino acid (AA5); (q) providing a sixth amino acid (AA6), and (r) coupling the sixth amino acid (AA6) with the fifth amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5-AA6) between the sixth amino acid (AA6) and the fifth amino acid; may include:

[0119] In some embodiments, the sixth amino acid (AA6) is:

[0120] [ka]

[0121] In some embodiments, the method of making a peptide or peptidomimetic comprises: (s) removing the protecting group from the sixth amino acid (AA6); (t) providing a seventh amino acid (AA7), and (u) coupling the seventh amino acid (AA6) with the sixth amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5-AA6-AA7) between the seventh amino acid (AA7) and the sixth amino acid; may include:

[0122] In some embodiments, the seventh amino acid (AA7) is:

[0123] [ka]

[0124] In some embodiments, the method of making a peptide or peptidomimetic comprises: (v) removing the protecting group from the seventh amino acid (AA7); (w) providing an eighth amino acid (AA8), and (x) coupling the eighth amino acid (AA8) with the seventh amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5-AA6-AA7-AA8) between the eighth amino acid (AA8) and the seventh amino acid; may include:

[0125] In some embodiments, the eighth amino acid (AA8) is:

[0126] [ka]

[0127] In some embodiments, the method of making a peptide or peptidomimetic comprises: (y) removing the protecting group from the eighth amino acid (AA8); (z) providing a ninth amino acid (AA9), and (aa) coupling the ninth amino acid (AA9) with the eighth amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5-AA6-AA7-AA8-AA9) between the ninth amino acid (AA9) and the eighth amino acid; may include:

[0128] In some embodiments, the ninth amino acid (AA9) is:

[0129] [ka]

[0130] In some embodiments, the method of making a peptide or peptidomimetic comprises: (bb) removing the protecting group from the ninth amino acid (AA9); (cc) providing a tenth amino acid (AA10), and (dd) coupling the tenth amino acid (AA10) with the ninth amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5-AA6-AA7-AA8-AA9-AA10) between the tenth amino acid (AA10) and the ninth amino acid; may include:

[0131] In one embodiment, the tenth amino acid (AA10) is:

[0132] [ka]

[0133] In some embodiments, the method of making a peptide or peptidomimetic comprises: (i) Amino acids (AA n removing the protecting group from (ii) the subsequent amino acid (AA n+1 ), and (iii) the subsequent amino acid (AA n+1 ) to amino acids (AA n ) to form the next amino acid (AA n+1 ) and amino acids (AA n forming a peptide bond between and repeating The repeating step may be performed 1 to 100 times.

[0134] In some embodiments, the method of making a peptide or peptidomimetic comprises: (ee) removing the protecting group from the tenth amino acid (AA10); (ff) Formula (B):

[0135] [ka] wherein x is an integer from 1 to 15; and (gg) coupling the compound of formula (B) with one of the amino acids in the peptide or peptidomimetic to form a triazole ring. may include:

[0136] In some embodiments, the compound of Formula (B) has the formula:

[0137] [ka]

[0138] In some embodiments, the method of making a peptide or peptidomimetic according to any of the above embodiments comprises: (hh) further cleaving the peptide bond between the first amino acid (AA1) and the compound of formula (I) to obtain a peptide or peptidomimetic.

[0139] In some embodiments, the first amino acid (AA1), the second amino acid (AA2), the third amino acid (AA3), the fourth amino acid (AA4), the fifth amino acid (AA5), the sixth amino acid (AA6), the seventh amino acid (AA7), the eighth amino acid (AA8), the ninth amino acid (AA9), the tenth amino acid (AA10), and any subsequent amino acids (AA n+1 ) are, respectively,

[0140] [ka] [In the formula, PG 5 are suitable protecting groups. In some embodiments, PG5 is independently selected at each occurrence from the group consisting of triphenylmethyl (Trt), allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

[0141] In some embodiments, the first amino acid (AA1), the second amino acid (AA2), the third amino acid (AA3), the fourth amino acid (AA4), the fifth amino acid (AA5), the sixth amino acid (AA6), the seventh amino acid (AA7), the eighth amino acid (AA8), the ninth amino acid (AA9), the tenth amino acid (AA10), and any subsequent amino acids (AA n+1 ) are, respectively,

[0142] [ka] are independently selected from the group consisting of:

[0143] In one embodiment, the peptidomimetic has the formula:

[0144] [ka] or a salt thereof.

[0145] In one embodiment, the peptidomimetic has the formula:

[0146] [ka] or a salt thereof.

[0147] In another embodiment, the compound of formula (AI):

[0148] [ka] [In the formula, m and n are independently an integer of 0 to 5; R1a is C 1~5 is alkyl, R 2a is, independently at each occurrence, optionally replaced by C 10~30 is alkyl, R 3a is, independently at each occurrence, optionally replaced by C 10~30 alkyl] Provided herein are compounds having the structure: or a salt thereof.

[0149] In some embodiments, R 1a is methyl.

[0150] In some embodiments, R 2a is, independently at each occurrence, optionally replaced by C 18~26 alkyl, and R 3a is, independently at each occurrence, optionally replaced by C 18~26 In some embodiments, each R 2a are independently optionally substituted C 22 alkyl, and each R 3a are independently optionally substituted C 22 It is alkyl.

[0151] In yet another embodiment, a compound of formula (BI):

[0152] [ka] [In the formula, m is an integer from 1 to 5; R 1b is C 1~5 is alkyl, R 2b is, independently at each occurrence, optionally replaced by C 10~30 alkyl] Provided herein are compounds having the structure: or a salt thereof.

[0153] In some embodiments, R 1b is methyl.

[0154] In some embodiments, R 2b is, independently at each occurrence, optionally replaced by C 18~26 In some embodiments, each R 2b are independently optionally substituted C 22 It is alkyl.

[0155] In yet another embodiment, a compound of formula (CI):

[0156] [ka] [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R 2c is, independently at each occurrence, optionally replaced by C 10~30 is alkyl, R 3c is, independently at each occurrence, optionally replaced by C 10~30 alkyl] Provided herein are compounds having the structure: or a salt thereof.

[0157] In some embodiments, R 2c is, independently at each occurrence, optionally replaced by C 18~26 alkyl, and R 3c is, independently at each occurrence, optionally replaced by C 18~26 In some embodiments, each R 2c are independently optionally substituted C 22 alkyl, and each R 3c are independently optionally substituted C 22 It is alkyl.

[0158] In yet another embodiment, a compound of formula (DI):

[0159] [ka] [In the formula, k is an integer from 1 to 15, q is an integer from 0 to 10, R 1d is C 1~6 is alkyl, PG 6 is a suitable protecting group, PG 7 is a suitable protecting group] or a salt thereof, the method comprising: (a) Structure:

[0160] [ka] providing a compound of formula (DII) having the formula: (b) forming a compound of formula (DI) from a compound of formula (DII); Includes:

[0161] In some embodiments, PG 6 is selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

[0162] In some embodiments, PG 7 may be selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

[0163] In some embodiments, the compound of Formula (DI) has the structure of Formula (DIa) or Formula (DIb).

[0164] [ka]

[0165] In some embodiments, the compound of Formula (DII) has the structure of Formula (DIIa) or Formula (DIIb).

[0166] [ka]

[0167] In some embodiments, step (b) of forming a compound of formula (DI) comprises: The compound of formula (DII) may be reacted with a compound of formula (DIII):

[0168] [ka] with a compound of formula (II) Includes:

[0169] In one embodiment, the compound of Formula (DIII) has the structure of Formula (DIIIa):

[0170] [ka]

[0171] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0172] [ka] or a salt thereof; and Before step (a), forming a compound of formula (DII) from a compound of formula (DIV). may include:

[0173] In some embodiments, the compound of formula (DIV) has the structure of formula (DIVa) or formula (DIVb).

[0174] [ka]

[0175] In some embodiments, the step of forming a compound of formula (DII) comprises: The compound of formula (DIV) is reacted with a compound of formula (DVa) PG 6 -X(DVa) or Formula (DVb) PG 6 2O(DVb) [In the formula, X is OSu, OTf, Cl, or Br, Producing a compound of formula (DII) Includes:

[0176] In some embodiments, the compound of Formula (DVa) or Formula (DVb) is selected from the group consisting of AllocCl, Alloc2O, Cbz2O, CbzCl, FmocCl, FmocOSu, and Boc2O.

[0177] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0178] [ka] [In the formula, PG 8 is a suitable protecting group; and forming a compound of formula (DIV) from a compound of formula (DVI) may include:

[0179] In some embodiments, the step of forming a compound of Formula (DIV) comprises reacting a compound of Formula (DVI) with a deprotecting agent to produce a compound of Formula (DIV). In some embodiments, the deprotecting agent may be selected from the group consisting of Pd(PPh)3, PhSiH3, H2, HCl, piperidine, and trifluoroacetic acid (TFA).

[0180] In some embodiments, the compound of Formula (DVI) has the structure of Formula (DVIa) or Formula (DVIb).

[0181] [ka]

[0182] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0183] [ka] providing a compound of formula (DVII) having the formula: forming a compound of formula (DVI) from a compound of formula (DVII) may include:

[0184] In some embodiments, the step of forming a compound of Formula (DVI) comprises reacting a compound of Formula (DVII) with a base to produce a compound of Formula (DVI). In some embodiments, the base is selected from the group consisting of LiOH, NaOH, and KOH.

[0185] In some embodiments, the compound of Formula (DVII) has the structure of Formula (DVIIa) or Formula (DVIIb).

[0186] [ka]

[0187] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0188] [ka] [In the formula, and LG′ is a suitable living group; and forming a compound of formula (DVII) from a compound of formula (DVIII) may include:

[0189] In some embodiments, the step of forming the compound of Formula (DVII) comprises reacting the compound of Formula (DVIII) with an azide group introducing agent to produce the compound of Formula (DVII). In some embodiments, the azide group introducing agent is NaN3, TMSN3, (PhO)2P(O)N3, Zn(N3)2 * 2Py, and n-Bu4NN3.

[0190] In some embodiments, LG' is selected from the group consisting of halogen, OTf, OMs, and OTs.

[0191] In some embodiments, the compound of Formula (DVIII) has the structure of Formula (DVIIIa) or Formula (DVIIIb).

[0192] [ka]

[0193] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0194] [ka] [In the formula, wherein LG″ is a suitable living group; and forming a compound of formula (DVIII) from a compound of formula (DIX) may include:

[0195] In some embodiments, LG″ is selected from the group consisting of halogen, OTf, OMs, and OTs.

[0196] In some embodiments, the step of forming a compound of formula (DVIII) comprises: The compound of formula (DIX) may be reacted with a compound of formula (DX):

[0197] [ka] [In the formula, PG 8 is a suitable protecting group, Producing a compound of formula (DVII) Includes:

[0198] In some embodiments, PG 8 may be selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc). 8 is tert-butyloxycarbonyl (Boc).

[0199] In some embodiments, the compound of Formula (DIX) has the structure of Formula (DIXa):

[0200] [ka]

[0201] In some embodiments, the compound of formula (DX) has the structure of formula (DXa).

[0202] [ka]

[0203] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0204] [ka] [In the formula, R 2d is C 1~6 wherein R is an alkylene; and forming a compound of formula (DIX) from a compound of formula (DXI) may include:

[0205] In some embodiments, the step of forming a compound of Formula (DIX) comprises reacting a compound of Formula (DXI) with a reducing agent to produce a compound of Formula (DIX). In some embodiments, the reducing agent is selected from the group consisting of H, NHHCO, NABH(OAc), and LiAlH.

[0206] In some embodiments, the compound of Formula (DXI) has the structure of Formula (DXIa):

[0207] [ka]

[0208] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0209] [ka] providing a compound of formula (DXII) having the formula: forming a compound of formula (DXI) from a compound of formula (DXII) may include:

[0210] In some embodiments, the step of forming a compound of formula (DXI) comprises: The compound of formula (DXII) may be reacted with a compound of formula (DXIII): PPh3 * C1~6 Alkyl-Hal(DXIII) [In the formula, where Hal is a halogen, Producing a compound of formula (DXI) Includes:

[0211] In some embodiments, the compound of Formula (DXII) has the structure of Formula (DXIIa).

[0212] [ka]

[0213] In some embodiments, the compound of Formula (DXIII) has the structure of Formula (DXIIIa): PPh3 * MeBr(DXIIIa)

[0214] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0215] [ka] providing a compound of formula (DXIV) having the formula: forming a compound of formula (DXII) from a compound of formula (DXIV) may include:

[0216] In some embodiments, the step of forming a compound of formula (DXII) comprises: The compound of formula (DXIV) may be reacted with a compound of formula (DXV):

[0217] [ka] [In the formula, wherein LG"' is a suitable living group, Producing a compound of formula (DXII) Includes:

[0218] In some embodiments, LG"' is selected from the group consisting of halogen, OTf, OMs, and OTs.

[0219] In some embodiments, the compound of Formula (DXIV) has the structure of Formula (DXIVa):

[0220] [ka]

[0221] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0222] [ka] [In the formula, LG * is a suitable living group; and forming a compound of formula (DX) from a compound of formula (DXVI) may include:

[0223] In some embodiments, LG * is selected from the group consisting of halogens, OTf, OMs, and OTs.

[0224] In some embodiments, the step of forming a compound of formula (DX) comprises: The compound of formula (DXVI) may be reacted with a compound of formula (DXVII):

[0225] [ka] with a compound of formula (DX) Includes:

[0226] In some embodiments, the compound of Formula (DXVI) has the structure of Formula (DXVIc):

[0227] [ka]

[0228] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0229] [ka] providing a compound of formula (DXVIII) having the formula: forming a compound of formula (DXVI) from a compound of formula (DXVIII) may include:

[0230] In some embodiments, the step of forming a compound of Formula (DXVI) comprises: The compound of formula (DXVIII) is reacted with a compound of formula (DXIXa) LG * -X(DXIXa) or Formula (DXIXb) LG * 2O(DXIXb) [In the formula, X is OSu, Cl, or Br, Producing a compound of formula (DXVI) Includes:

[0231] In some embodiments, the compound of Formula (DXIXa) or Formula (DXIXb) is selected from the group consisting of TfCl, Tf2O, MsCl, and Ms2O.

[0232] In a further embodiment, the compound of formula (DII):

[0233] [ka] [In the formula, q is an integer from 0 to 10, R 1d is C 1~6 is alkyl, PG 6 is a suitable protecting group] or a salt thereof, said process comprising: (a) Structure:

[0234] [ka] or a salt thereof; and (b) forming a compound of formula (DII) from a compound of formula (DXXI); A method is provided herein, comprising:

[0235] In some embodiments, PG 6 is selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

[0236] In some embodiments, the compound of formula (DXXI) is treated with the following compound:

[0237] [ka]

[0238] In some embodiments, the compound of Formula (DXXI) is treated with a catalyst. In some embodiments, the catalyst is Pd(dppf)Cl-DCM.

[0239] In some embodiments, the compound of Formula (DXXI) is treated with a base. In some embodiments, the base is K2CO3.

[0240] In some embodiments, the preparation is carried out in a solvent mixture composed of H2O / iPrOH.

[0241] In some embodiments, the compound of Formula (DII) has the structure of Formula (DIIa) or Formula (DIIb).

[0242] [ka]

[0243] In some embodiments, the compound of formula (DXXI) is

[0244] [ka] of

[0245] [ka] It is formed by exposure to

[0246] In some embodiments, the compound of Formula (DXXI) is formed in the presence of PPh3.

[0247] In some embodiments, the compound of Formula (DXXI) is formed in the presence of DIAD.

[0248] In some embodiments, the compound of formula (DXXI) is formed in the presence of 4 Å molecular sieves.

[0249] In some embodiments, the compound of Formula (DXXI) is formed in an organic solvent. In some embodiments, the organic solvent is THF.

[0250] In some embodiments,

[0251] [ka] teeth,

[0252] [ka] is formed by exposing it to PBin2.

[0253] In some embodiments,

[0254] [ka] is formed in the presence of Pd(OAc)2.

[0255] In some embodiments,

[0256] [ka] is formed in the presence of PCy3.

[0257] In some embodiments,

[0258] [ka] is formed in the presence of KOAc.

[0259] In some embodiments,

[0260] [ka] is formed in the presence of dioxane.

[0261] In some embodiments,

[0262] [ka] is formed at elevated temperatures over a period of time.

[0263] In some embodiments,

[0264] [ka] teeth,

[0265] [ka] Bu4N + Br3 - It is formed by exposure to

[0266] In some embodiments,

[0267] [ka] is formed in an organic solvent. In some embodiments, the organic solvent is a mixture of DCM and methanol.

[0268] In a further embodiment, the compound of formula (DII):

[0269] [ka] [In the formula, q is an integer from 0 to 10, R 1d is C 1~6 is alkyl, PG 6 is a suitable protecting group] or a salt thereof, said process comprising: (a) Structure:

[0270] [ka] or a salt thereof; and (b) forming a compound of formula (DII) from a compound of formula (DIV); A method is provided herein, comprising:

[0271] In some embodiments, PG 6may be selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

[0272] In some embodiments, the compound of Formula (DII) has the structure of Formula (DIIa) or Formula (DIIb).

[0273] [ka]

[0274] In some embodiments, the compound of formula (DIV) has the structure of formula (DIVa) or formula (DIVb).

[0275] [ka]

[0276] In some embodiments, the process for forming the compound of Formula (DII) comprises: The compound of formula (DIV) is reacted with a compound of formula (DVa) PG 6 -X(DVa) or Formula (DVb) PG 6 2O(DVb) [In the formula, X is OSu, OTf, Cl, or Br, Producing a compound of formula (DII) Includes:

[0277] In some embodiments, the compound of Formula (DVa) or Formula (DVb) is selected from the group consisting of AllocCl, Alloc2O, Cbz2O, CbzCl, FmocCl, FmocOSu, and Boc2O.

[0278] In some embodiments, the process for preparing a compound of formula (DII) comprises: structure:

[0279] [ka] [In the formula, PG 8 is a suitable protecting group; and forming a compound of formula (DIV) from a compound of formula (DXVI) may include:

[0280] In some embodiments, PG 8 is selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

[0281] In some embodiments, the step of forming a compound of Formula (DIV) comprises reacting a compound of Formula (DXVI) with a deprotecting agent to produce a compound of Formula (DIV). In some embodiments, the deprotecting agent is selected from the group consisting of HCl, Pd(PPh), PhSiH, H, piperidine, and trifluoroacetic acid (TFA).

[0282] In some embodiments, the compound of Formula (DXVI) has the structure of Formula (DXVIa) or Formula (DXVIb).

[0283] [ka]

[0284] In some embodiments, the process for preparing a compound of formula (DII) comprises: structure:

[0285] [ka] [In the formula, and LG′ is a suitable living group; and forming a compound of formula (DXVI) from a compound of formula (DXVII) may include:

[0286] In some embodiments, the step of forming a compound of formula (DXVI) comprises reacting a compound of formula (DXVII) with an azide group introducing agent to produce a compound of formula (DXVI). In some embodiments, the azide group introducing agent is NaN3, TMSN3, (PhO)2P(O)N3, Zn(N3)2 * 2Py, and n-Bu4NN3.

[0287] In some embodiments, LG' is selected from the group consisting of halogen, OTf, OMs, and OTs.

[0288] In some embodiments, the compound of formula (DXVII) has the structure of formula (DXVIIa) or formula (DXVIIb).

[0289] [ka]

[0290] In some embodiments, the process for preparing a compound of formula (DII) comprises: structure:

[0291] [ka] [In the formula, wherein LG″ is a suitable living group; and forming a compound of formula (DXVII) from a compound of formula (DIX) may include:

[0292] In some embodiments, LG″ is selected from the group consisting of halogen, OTf, OMs, and OTs.

[0293] In some embodiments, the step of forming a compound of formula (DXVII) comprises: The compound of formula (DIX) may be reacted with a compound of formula (DXVIII):

[0294] [ka] with a compound of formula (DXVII) Includes:

[0295] In some embodiments, the compound of Formula (DIX) has the structure of Formula (DIXa):

[0296] [ka]

[0297] In some embodiments, the compound of Formula (DXVIII) has the structure of Formula (DXVIIIa):

[0298] [ka]

[0299] In some embodiments, the process for preparing a compound of formula (DII) comprises: structure:

[0300] [ka] providing a compound of formula (DXIX) having the formula: forming a compound of formula (DIX) from a compound of formula (DXIX) may include:

[0301] In some embodiments, the step of forming a compound of formula (DIX) comprises: The compound of formula (DXIX) may be reacted with a compound of formula (DXV):

[0302] [ka] [In the formula, wherein LG"' is a suitable living group, Producing a compound of formula (DXIX) Includes:

[0303] In some embodiments, LG"' is selected from the group consisting of halogen, OTf, OMs, and OTs.

[0304] In some embodiments, the compound of Formula (DXIX) has the structure of Formula (DXIXa):

[0305] [ka]

[0306] In some embodiments, the compound of formula (DXV) has the structure of formula (DXVa):

[0307] [ka]

[0308] In some embodiments, the process for preparing a compound of formula (DII) comprises: structure:

[0309] [ka] providing a compound of formula (DXX) having the formula: forming a compound of formula (DXIX) from a compound of formula (DXX) may include:

[0310] In some embodiments, The step of forming a compound of formula (DXIX) comprises: reacting a compound of formula (DXX) with an LG"-introducing agent to produce a compound of formula (DXIX); Includes:

[0311] In some embodiments, the LG″ introducing agent is NBu4Br3.

[0312] In some embodiments, the compound of Formula (DXX) has the structure of Formula (DXXa).

[0313] [ka]

[0314] In another aspect, provided herein is a pharmaceutical composition comprising a peptidomimetic or compound of any of the embodiments described herein.

[0315] In another aspect, provided herein is a pharmaceutical dosage form comprising a peptidomimetic or compound of any of the embodiments described herein.

[0316] In another aspect, the present disclosure provides a method for selectively targeting GLP1R on the surface of a cell by using any of the peptidomimetics or compounds of the embodiments described herein.In some embodiments, the cell is a mammalian cell.In some embodiments, the cell is a human cell.In some embodiments, the cell is a pancreatic cell, a brain cell, a cardiac cell, a vascular tissue cell, a kidney cell, an adipose tissue cell, a liver cell, or a muscle cell.

[0317] In another aspect, provided herein is a method of enhancing activity of GLP1R in an individual in need thereof, comprising administering to the individual an effective amount of a peptidomimetic or compound of any of the embodiments described herein, a composition described herein, or a dosage form described herein.

[0318] In another aspect, provided herein is a method of lowering blood glucose levels in an individual in need thereof, comprising administering to the individual an effective amount of a peptidomimetic or compound of any of the embodiments described herein, a composition described herein, or a dosage form described herein.

[0319] In another aspect, provided herein is a method of reducing body weight in an individual in need thereof, comprising administering to the individual an effective amount of a peptidomimetic or compound of any of the embodiments described herein, a composition described herein, or a dosage form described herein.

[0320] In another aspect, provided herein is a method for treating a GLP1R-related disease in an individual in need thereof, comprising administering to the individual an effective amount of any of the peptidomimetics or compounds of the embodiments described herein, the compositions described herein, or the dosage forms described herein. In some embodiments, the GLP1R-related disease is type II diabetes, obesity, liver disease, coronary artery disease, or kidney disease. In some embodiments, the GLP1R-related disease is type II diabetes and / or obesity.

[0321] In various embodiments of any of the methods described herein, the disclosed compounds, compositions, or dosage forms are administered subcutaneously, intravenously, intradermally, intraperitoneally, or intramuscularly.

[0322] These and other aspects of the present disclosure will become apparent to those skilled in the art upon reading the following detailed description of the present disclosure, including the appended claims.

[0323] The patent or application file contains at least one color drawing. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee. [Brief explanation of the drawings]

[0324] [Figure 1-1] FIG. 1 shows a liquid phase peptide support (LPPS) approach to making M1. [Figure 1-2] FIG. 1 shows a liquid phase peptide support (LPPS) approach to making M1. [Figure 1-3] FIG. 1 shows a liquid phase peptide support (LPPS) approach to making M1. [Figure 2-1] FIG. 1 shows the synthesis of Tag1-M2. [Figure 2-2] FIG. 1 shows the synthesis of Tag1-M2. [Figure 2-3] FIG. 1 shows the synthesis of Tag1-M2. [Figure 3-1] FIG. 1 shows the synthesis of Tag2-M2. [Figure 3-2] FIG. 1 shows the synthesis of Tag2-M2. [Figure 3-3] FIG. 1 shows the synthesis of Tag2-M2. [Figure 4-1] FIG. 1 shows the synthesis of tag 3-M2. [Figure 4-2] FIG. 1 shows the synthesis of tag 3-M2. [Figure 4-3] FIG. 1 shows the synthesis of tag 3-M2. [Figure 4-4] FIG. 1 shows the synthesis of tag 3-M2. [Figure 5-1] FIG. 1 shows the synthesis of tag 4-M2. [Figure 5-2] FIG. 1 shows the synthesis of tag 4-M2. [Figure 5-3] FIG. 1 shows the synthesis of tag 4-M2. [Figure 5-4] FIG. 1 shows the synthesis of tag 4-M2. [Figure 6-1] FIG. 1 shows the synthesis of tag 5-M1. [Figure 6-2] FIG. 1 shows the synthesis of tag 5-M1. [Figure 6-3] FIG. 1 shows the synthesis of tag 5-M1. [Figure 6-4] FIG. 1 shows the synthesis of tag 5-M1. [Figure 6-5]FIG. 1 shows the synthesis of tag 5-M1. [Figure 6-6] FIG. 1 shows the synthesis of tag 5-M1. [Figure 7-1] FIG. 1 shows the synthesis of tag 5-M3. [Figure 7-2] FIG. 1 shows the synthesis of tag 5-M3. [Figure 7-3] FIG. 1 shows the synthesis of tag 5-M3. [Figure 7-4] FIG. 1 shows the synthesis of tag 5-M3. [Figure 7-5] FIG. 1 shows the synthesis of tag 5-M3. [Figure 7-6] FIG. 1 shows the synthesis of tag 5-M3. [Figure 7-7] FIG. 1 shows the synthesis of tag 5-M3. [Figure 8-1] FIG. 1 shows some examples of known LPPS tags. [Figure 8-2] FIG. 1 shows some examples of known LPPS tags. [Figure 8-3] FIG. 1 shows some examples of known LPPS tags. [Figure 9] FIG. 1 shows SPPS for peptide elongation. [Figure 10A] 1 is a table showing a summary of the procedures and conditions used to prepare M1 using Tag 5. [Figure 10B] 1 is a table showing a summary of the procedures and conditions used to prepare M1 using Tag 5. [Figure 11A] 1 is a table showing the extension conditions used to generate M1 using tag 5 and the results. [Figure 11B] 1 is a table showing the extension conditions used to generate M1 using tag 5 and the results. [Figure 12] 1 is a table showing the extension conditions used to generate M3 using tag 5 and the results. [Figure 13-1] 1 is a table showing a comparison of five routes used to prepare AA2 and AA2+ linkers. [Figure 13-2]1 is a table showing a comparison of five routes used to prepare AA2 and AA2+ linkers. [Figure 14-1] FIG. 1 shows the synthesis of AA2 using Route 1. [Figure 14-2] FIG. 1 shows the synthesis of AA2 using Route 1. [Figure 14-3] FIG. 1 shows the synthesis of AA2 using Route 1. [Figure 15] FIG. 1 shows an alternative process for generating AA2 and AA2+ linkers. DETAILED DESCRIPTION OF THE INVENTION

[0325] In some embodiments, the present disclosure provides compounds useful as supports for solution-phase organic synthesis. Several novel LPPS tags (Tag 2 to Tag 5 in Figure 1) were designed by a) increasing the hydrophobicity of the tag for solidification of TCP constructs to remove residual reactants and reagents and impurities in solution for easy separation and purification, and b) introducing a 2,4-bismethoxy-benzylamine amide moiety (e.g., Tag 5) for easy C-N bond cleavage. After C->N synthesis of Fmoc / tBu, GLP1 peptide mimetics were prepared on the new tags.

[0326] A new liquid-phase peptide support (LPPS) tag was designed that combines the benefits of solid-phase peptide support (SPPS) synthesis, which allows separation of the solid-supported peptide from the extension mixture, with the benefits of traditional solution peptide synthesis (CSPS), which uses fewer molar equivalents of reactants and reagents to grow the peptide chain. The increased hydrophobicity of the LPPS tag allowed i) more steps of peptide extension on a soluble tag in the extension solvent, and ii) precipitation of the tagged supported / growing peptide in polar solvents. Tag 5, with a six-member long alkyl chain, allowed LPPS of the hydrophilic GLP1 agonist to use fewer molar equivalents of reactants and reagents compared to SPPS synthesis.

[0327] Although detailed embodiments of the present disclosure are disclosed herein, it should be understood that the disclosed embodiments are merely exemplary of the present disclosure, which may be embodied in various forms. In addition, each example presented in connection with various embodiments of the present disclosure is illustrative and not intended to be limiting. Therefore, the specific structural and functional details disclosed herein should not be construed as limiting, but merely representative examples based on teachings to those skilled in the art who may utilize the present disclosure in various ways.

[0328] definition 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.

[0329] As used in this application and the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, reference to "a method" includes one or more methods, and / or steps of the type described herein and / or that will become apparent to those skilled in the art upon reading this disclosure.

[0330] "Subject" or "patient" or "individual" or "animal," as used herein, refers to humans, domestic animals (e.g., cats, dogs, cows, horses, sheep, pigs, etc.), and experimental animal models of disease (e.g., mice, rats). In a preferred embodiment, the subject is a human.

[0331] The phrase "pharmaceutically acceptable salt" when used in connection with the compositions of the present disclosure refers to any salt suitable for administration to a patient. Suitable salts include, but are not limited to, those described in Berge et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977, 66:1, which is incorporated herein by reference. Examples of salts include, but are not limited to, calcium salts, magnesium salts, potassium salts, sodium salts, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, para-toluenesulfonic acid, salicylic acid, and the like, and include, but are not limited to, acid-derived, base-derived, organic, inorganic, amine, and alkali or alkaline earth metal salts.

[0332] Ranges may be expressed herein as from "about" or "approximately" one particular value and / or to "about" or "approximately" another particular value. When such a range is expressed, another embodiment includes from the one particular value and / or to the other particular value.

[0333] "Comprising" or "containing" or "including" means that at least the named compounds, elements, particles, or method steps are present in the composition or article or method, but does not exclude the presence of other compounds, materials, particles, or method steps, even if they have the same function as the named ones.

[0334] Compounds of the present disclosure, such as tags, include those generally described herein, and are further exemplified by the classes, subclasses, and species disclosed herein.As used herein, the following definitions apply unless otherwise indicated.For the purpose of this disclosure, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th edition.In addition, the general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5th edition, Smith, MB and March, J. (eds.), John Wiley & Sons, New York: 2001, which are incorporated herein by reference in their entirety.

[0335] As used herein, the term "alkyl" is given its ordinary meaning in the art and can include saturated aliphatic groups, including straight-chain and branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups. In certain embodiments, a straight-chain or branched-chain alkyl has from about 1 to 30 carbon atoms in its backbone (e.g., C1 to C6 for a straight chain). 30 , C2 to C for branched chains 30 ), alternatively having about 1-10 carbon atoms, or about 1-6 carbon atoms. In some embodiments, cycloalkyl rings have from about 3-10 carbon atoms in their ring structure, and such rings are monocyclic or bicyclic, alternatively having about 5, 6 or 7 carbons in the ring structure. In some embodiments, alkyl groups can be lower alkyl groups, which lower alkyl groups have from 1-5 carbon atoms (e.g., C1-C5 for a straight chain lower alkyl).

[0336] The term "halogen" means F, Cl, Br, or I, and the term "halide" refers to a halogen radical or substituent, i.e., -F, -Cl, -Br, or -I.

[0337] As described herein, compounds (e.g., tags) of the present disclosure may contain "optionally substituted" moieties. Generally, the term "substituted," whether preceded by the term "optionally," means that one or more hydrogen atoms in the specified moiety have been replaced with a suitable substituent. For example, up to three H atoms in each residue are replaced with alkyl, halogen, haloalkyl, hydroxy, lower alkoxy, carboxy, carboalkoxy (also referred to as alkoxycarbonyl), carboxamido (also referred to as alkylaminocarbonyl), cyano, carbonyl, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, sulfoxide, sulfone, acylamino, amidino, phenyl, benzyl, heteroaryl, phenoxy, benzyloxy, or heteroaryloxy. Unless otherwise indicated, an "optionally substituted" group can have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure can be substituted with more than one substituent selected from a specified group, the substituents can be the same or different at all positions. Combinations of substituents envisioned by this disclosure are preferably those that result in the formation of stable or chemically feasible compounds.

[0338] The term "stable," as used herein with reference to a compound, refers to a compound that is not substantially altered when subjected to conditions that allow for its production, detection, and, in certain embodiments, recovery, purification, and use for one or more purposes disclosed herein.

[0339] Unless otherwise stated, structures depicted herein are also intended to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure, such as the R and S configurations at each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) stereoisomers. Accordingly, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the disclosure.

[0340] Unless otherwise stated, the cycloadducts depicted herein, e.g., products of a cycloaddition reaction, e.g., an azide acetylene cycloaddition reaction or a Diels-Alder reaction, include all regioisomers, i.e., structural isomers that differ only in the position of functional groups or substituents. By way of example, the following structures represent triazole regioisomers, which differ only in the position of substituents on the triazole ring:

[0341] [ka] The triazole positional isomers may also be represented by the following structures:

[0342] [ka]

[0343] Unless otherwise stated, all tautomeric forms of the compounds of the present disclosure are within the scope of the present disclosure.

[0344] Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms, for example, the replacement of hydrogen by deuterium or tritium, or 11 C- or 13 C- or 14 Compounds having the present structures except for the replacement of a carbon with a C-enriched carbon are within the scope of this disclosure.

[0345] It should also be understood that the reference to one or more method steps does not exclude the presence of additional or intervening method steps between the explicitly identified steps. Similarly, it should also be understood that the reference to one or more components in a device or system does not exclude the presence of additional or intervening components between the explicitly identified components.

[0346] Unless otherwise stated, all crystalline forms of the compounds of the present disclosure and their salts are also within the scope of the present disclosure.The compounds of the present disclosure can be isolated in various amorphous and crystalline forms, including, but not limited to, anhydrous, hydrated, non-solvated, or solvated forms.Examples of hydrates include hemihydrate, monohydrate, dihydrate, etc.In some embodiments, the compounds of the present disclosure are anhydrous and non-solvated."Anhydrous" means that the crystalline form of the compound essentially does not contain bound water in the crystal lattice structure, i.e., the compound does not form crystalline hydrate.

[0347] As used herein, "crystalline form" is intended to refer to a particular lattice configuration of a crystalline substance. Different crystalline forms of the same substance typically have different crystal lattices (e.g., unit cells), which contribute to the different physical properties characteristic of each crystalline form. In some cases, different lattice configurations have different water or solvent contents. Different crystal lattices can be confirmed by solid-state characterization methods such as X-ray powder diffraction (PXRD). Other characterization methods, such as differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), dynamic vapor sorption (DVS), solid-state NMR, etc., further aid in the identification of crystalline forms and also aid in the determination of stability and solvent / water content.

[0348] Crystalline forms of a substance include both solvated (e.g., hydrated) and non-solvated (e.g., anhydrous) forms. Hydrated forms are crystalline forms that contain water in the crystal lattice. Hydrated forms may be stoichiometric hydrates, in which water is present in the lattice at a specific water / molecule ratio, e.g., hemihydrate, monohydrate, dihydrate, etc. Hydrated forms may also be non-stoichiometric, in which the amount of water varies and is dependent on external conditions such as humidity.

[0349] In some embodiments, the compounds of the present disclosure are substantially isolated. "Substantially isolated" means that a particular compound is at least partially isolated from impurities. For example, in some embodiments, the compounds of the present disclosure contain less than about 50%, less than about 40%, less than about 30%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, less than about 2.5%, less than about 1%, or less than about 0.5% impurities. Impurities generally include any of the compounds that are not substantially isolated, including, for example, other crystalline forms and other substances.

[0350] The term "GLP1R" refers to glucagon-like peptide 1 receptor and includes recombinant GLP1R protein or fragments thereof. GLP1R has a sequence of 463 residues. Donnelly, Br J Pharmacol, 166(1):27-41 (2011). Glucagon-like peptide 1 (GLP1) is a 31-amino acid peptide hormone released from small intestinal L-cells after nutrient consumption. Binding of GLP1 to GLP1R enhances glucose-induced insulin secretion from pancreatic beta cells, increases insulin expression, inhibits beta cell apoptosis, promotes beta cell neogenesis, reduces glucagon secretion, delays gastric emptying, promotes satiety, and increases peripheral glucose clearance.

[0351] tag In accordance with the above objectives and others, the present disclosure provides compounds that can be used as tags for the preparation of GLP1 peptide mimetics, as well as precursors and intermediates thereof. The present disclosure also provides methods for making these compounds, as well as precursors and intermediates thereof.

[0352] Also provided in the present disclosure are GLP1 peptide mimetics prepared using the tags described herein, and pharmaceutical compositions comprising the GLP1 peptide mimetics, as well as methods for treating certain diseases in subjects in need of such treatment.

[0353] In one aspect, the present disclosure provides a compound of formula (I):

[0354] [ka] [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R 1 is C 1~5 is alkyl, R 2 is, independently at each occurrence, optionally replaced by C 10~30 is alkyl, R 3 is, independently at each occurrence, optionally replaced by C 10~30 alkyl] The present invention provides a compound having the structure:

[0355] According to the present disclosure, m can be an integer from 1 to 5, and n can be an integer from 1 to 5, provided that (m+n) > 3. For example, m can be 1, 2, 3, 4, or 5, and n can be 1, 2, 3, 4, or 5.

[0356] In at least one embodiment, m is 3 and n is 3.

[0357] According to this disclosure, R 1 can be methyl, ethyl, propyl, butyl, or pentyl. For example, R 1 can be n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, or n-pentyl. 1 is methyl.

[0358] According to this disclosure, R 2 is, independently at each occurrence, optionally replaced by C 10~30 It is an alkyl group. For example, C 10~30 The alkyl may be optionally substituted 1 to 10 times with a substituent selected from the group consisting of halogen, hydroxy, lower alkoxy, carboxy, carboalkoxy (also called alkoxycarbonyl), carboxamido (also called alkylaminocarbonyl), cyano, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, phenyl, benzyl, heteroaryl, phenoxy, or benzyloxy. 10~30 The alkyl may be optionally substituted 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times with any of the above substituents, each substituent independently selected from halogen, hydroxy, lower alkoxy, carboxy, carboalkoxy, carboxamido, cyano, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, phenyl, benzyl, heteroaryl, phenoxy, or benzyloxy.

[0359] According to this disclosure, R 3 is, independently at each occurrence, optionally replaced by C 10~30 It is an alkyl group. For example, C 10~30 Alkyl may be optionally substituted 1 to 10 times with substituents selected from halogen, hydroxy, lower alkoxy, carboxy, carboalkoxy (also called alkoxycarbonyl), carboxamido (also called alkylaminocarbonyl), cyano, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, phenyl, benzyl, heteroaryl, phenoxy, or benzyloxy. For example, C 10~30The alkyl may be optionally substituted 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times with any of the above substituents, each substituent independently selected from halogen, hydroxy, lower alkoxy, carboxy, carboalkoxy, carboxamido, cyano, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, phenyl, benzyl, heteroaryl, phenoxy, or benzyloxy.

[0360] In at least one embodiment, R 2 is, independently at each occurrence, optionally replaced by C 18~26 alkyl, and R 3 is, independently at each occurrence, optionally replaced by C 18~26 For example, each R 2 are independently optionally substituted C 22 alkyl, and each R 3 are independently optionally substituted C 22 It is alkyl.

[0361] In one embodiment, a compound of formula (Ia):

[0362] [ka] Provided herein are compounds having the structure: or a salt thereof.

[0363] In another aspect, the present disclosure provides a compound of formula (I):

[0364] [ka] [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R 1 is C 1~5 is alkyl, R 2 is, independently at each occurrence, optionally replaced by C 10~30 is alkyl, R3 is, independently at each occurrence, optionally replaced by C 10~30 alkyl] or a salt thereof, the process comprising: (a) Structure:

[0365] [ka] [In the formula, wherein PG is a suitable protecting group; and (b) forming a compound of formula (I) from a compound of formula (II); Includes:

[0366] According to the present disclosure, PG is any suitable protecting group that can be used to protect an amino group. Suitable protecting groups that can be used are known to those skilled in the art. A detailed description of these groups and their selection and chemistry is contained in "The Peptides, Vol. 3," edited by Gross and Meinenhofer, Academic Press, New York, 1981, which is incorporated herein by reference in its entirety. For example, PG can be selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc). In some embodiments, PG is allyloxycarbonyl (Alloc).

[0367] In some embodiments, the compound of Formula (II) has the structure of Formula (IIa):

[0368] [ka]

[0369] In some embodiments, step (b) of forming a compound of Formula (I) may include reacting a compound of Formula (II) with a deprotecting agent to produce a compound of Formula (I).

[0370] According to the present disclosure, any suitable deprotecting agent can be used to remove the protecting group PG from the amino group of the compound of formula (II). Suitable deprotecting agents that can be used to remove specific protecting groups are known to those skilled in the art. A detailed description of these groups and their selection and chemistry is contained in "The Peptides, Vol. 3," edited by Gross and Meinenhofer, Academic Press, New York, 1981, which is incorporated herein by reference in its entirety. For example, the deprotecting agent can be selected from the group consisting of Pd(PPh)3, PhSiH3, H2, piperidine, and trifluoroacetic acid (TFA).

[0371] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0372] [ka] [In the formula, PG 1 is a suitable protecting group, PG 2 is a suitable protecting group; and Before step (a), forming a compound of formula (II) from a compound of formula (III). may include:

[0373] According to this disclosure, PG 1 is any suitable protecting group that can be used to protect a hydroxy group. Suitable protecting groups that can be used are known to those skilled in the art and can be selected from those described in "Handbook of Reagents for Organic Synthesis. Activating Reagents and Protecting Groups," edited by Pearson and Roush, John Wiley & Sons, 2005, which is incorporated herein by reference in its entirety. For example, PG1 may be selected from the group consisting of tert-butyldimethylsilyl (TBS), trimethylsilyl (TMS), triethylsilyl (TES), tert-butyldiphenylsilyl (TBDPS), and triisopropylsilyl (TIPS).

[0374] According to this disclosure, PG 2 is any suitable protecting group that can be used to protect a hydroxy group. Suitable protecting groups that can be used are known to those skilled in the art and can be selected from those described in "Handbook of Reagents for Organic Synthesis. Activating Reagents and Protecting Groups," edited by Pearson and Roush, John Wiley & Sons, 2005, which is incorporated herein by reference in its entirety. For example, PG 2 may be selected from the group consisting of tert-butyldimethylsilyl (TBS), trimethylsilyl (TMS), triethylsilyl (TES), tert-butyldiphenylsilyl (TBDPS), and triisopropylsilyl (TIPS).

[0375] According to this disclosure, each PG 1 and each PG 2 may be the same or different. In some embodiments, PG 1 is PG 2 For example, if m is 3 and n is 3, then three PG 1 The groups are identical, and the three PG 2 The groups are identical, and 1 =PG 2 In some embodiments, PG 1 is TBS and PG 2 is TBS. Or PG 1 and P.G. 2 For example, when m is 3 and n is 3, the three PGs 1 Group (PG 11 , P.G. 12 , and P.G.13 ) are the same (PG 1 =PG 11 =PG 12 =PG 13 ) and three PG 2 Group (PG 21 , P.G. 22 , and P.G. 23 ) are the same (PG 2 =PG 21 =PG 22 =PG 23 ), but PG 1 and P.G. 2 are different. In some embodiments, PG 1 is TBS and PG 2 is a TIPS. Or, each PG 1 and each PG 2 For example, when m is 3 and n is 3, the three PGs 1 Each of the groups is different from each other (PG 11 is PG 12 Unlike PG 12 is PG 13 Unlike PG 11 is PG 13 (different from the above), 3 PG 2 The groups are the same (PG 21 is PG 22 Unlike PG 22 is PG 23 Unlike PG 21 is PG 23 (Different from PG) 11 , P.G. 12 , P.G. 13 , P.G. 21 , P.G. 22 , and P.G. 23 There are no identical ones.

[0376] In some embodiments, the compound of Formula (III) has the structure of Formula (IIIa):

[0377] [ka] In some embodiments, the step of forming a compound of formula (II) prior to step (a) comprises: reacting a compound of formula (III) with a deprotecting agent to obtain a deprotected intermediate; The intermediate is reacted with the compound of formula (IV): R-LG(IV) [In the formula, R is an optionally substituted C 10~30 is alkyl, wherein LG is a suitable leaving group, Producing a compound of formula (II) Includes:

[0378] According to the present disclosure, any suitable deprotecting agent can be used to remove the protecting group PG from the hydroxyl group of the compound of formula (III). 1 and P.G. 2 Suitable protecting group removing agents that can be used to remove a particular protecting group are known to those skilled in the art and can be selected from those described in "Handbook of Reagents for Organic Synthesis. Activating Reagents and Protecting Groups," edited by Pearson and Roush, John Wiley & Sons, 2005, which is incorporated herein by reference in its entirety. For example, the protecting group removing agent can be selected from the group consisting of tetrabutylammonium fluoride (TBAF), NHF, SiF, AcOH, HCl, LiAlH, and KCO.

[0379] In some embodiments, PG 1 and P.G. 2 are different from each other, different deprotecting agents can be used. In this case, one deprotecting agent can be used to remove all PG present in the compound of formula (III). 1 The intermediate formed can then be reacted with a compound of formula (IVa), which is the first compound of formula (IV). Then, a separate protecting group removing agent can be used to selectively remove all of the PG groups. 2The group can be selectively removed and the deprotected hydroxyl group can then be modified using another compound of formula (IV), a compound of formula (IVb), where the compound of formula (IVa) can be different from the compound of formula (IVb).

[0380] According to the present disclosure, LG is a suitable leaving group. Suitable leaving groups that can be used are known to those skilled in the art. For example, LG can be selected from the group consisting of halogen, OTf, OMs, and OTs.

[0381] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0382] [ka] providing a compound of formula (V) having the formula: forming a compound of formula (III) from a compound of formula (V); may include:

[0383] In some embodiments, the step of forming a compound of formula (III) comprises: Compounds of formula (V) can be reacted with compounds of formula (VIa) PG-X(VIa) or Formula (VIb) PG2O(VIb) [In the formula, X is Cl or Br, Producing a compound of formula (III) may include:

[0384] According to the present disclosure, suitable compounds of formula (VIa) or formula (VIb) may be selected from the group consisting of, but not limited to, AllocCl, Alloc2O, Cbz2O, CbzCl, FmocCl, and Boc2O.

[0385] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0386] [ka] [In the formula, PG 3 is a suitable protecting group; and forming a compound of formula (V) from a compound of formula (VII) may include:

[0387] According to this disclosure, PG 3 is any suitable protecting group that can be used to protect an amino group. Suitable protecting groups that can be used are known to those skilled in the art. A detailed description of these groups, as well as their selection and chemistry, is contained in "The Peptides, Vol. 3," edited by Gross and Meinenhofer, Academic Press, New York, 1981, which is incorporated herein by reference in its entirety. For example, PG 3 may be selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc). 3 is 9-fluorenylmethoxycarbonyl (Fmoc).

[0388] In some embodiments, the compound of formula (VII) has the following structure:

[0389] [ka]

[0390] In some embodiments, the process for preparing a compound of formula (V) comprises reacting a compound of formula (VII) with a deprotecting agent to produce a compound of formula (V).

[0391] According to the present disclosure, any suitable deprotecting agent can be used to remove the protecting group PG from the amino group of the compound of formula (VII).3 Suitable protecting group removing agents that can be used to remove specific protecting groups are known to those skilled in the art. A detailed description of these groups, as well as their selection and chemistry, is contained in "The Peptides, Vol. 3," edited by Gross and Meinenhofer, Academic Press, New York, 1981, which is incorporated herein by reference in its entirety. For example, the protecting group removing agent is selected from the group consisting of Pd(PPh)3, PhSiH3, H2, piperidine, and trifluoroacetic acid (TFA).

[0392] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0393] [ka] providing a compound of formula (VIII) having the formula: forming a compound of formula (VII) from a compound of formula (VIII) may include:

[0394] In some embodiments, the process for forming a compound of Formula (VII) comprises reacting a compound of Formula (VIII) with the structure:

[0395] [ka] with a compound of formula (IX) having the formula (VII)

[0396] In one embodiment, the compound of formula (VIII) has the following structure:

[0397] [ka]

[0398] In another embodiment, the compound of formula (IX) has the structure:

[0399] [ka]

[0400] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0401] [ka] [In the formula, PG 4 is a suitable protecting group; and forming a compound of formula (VIII) from a compound of formula (X); may include:

[0402] According to this disclosure, PG 4 is any suitable protecting group that can be used to protect an amino group. Suitable protecting groups that can be used are known to those skilled in the art. A detailed description of these groups, as well as their selection and chemistry, is contained in "The Peptides, Vol. 3," edited by Gross and Meinenhofer, Academic Press, New York, 1981, which is incorporated herein by reference in its entirety. For example, PG 4 may be selected from the group consisting of allyl, allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc). 4 is an allyl.

[0403] In one embodiment, the compound of formula (X) has the structure:

[0404] [ka]

[0405] In some embodiments, the process of forming a compound of Formula (VIII) comprises reacting a compound of Formula (X) with a deprotecting agent.

[0406] According to the present disclosure, any suitable deprotecting agent can be used to remove the protecting group PG from the amino group of the compound of formula (X). 4 Suitable protecting group removing agents that can be used to remove specific protecting groups are known to those skilled in the art. A detailed description of these groups, as well as their selection and chemistry, is contained in "The Peptides, Vol. 3," edited by Gross and Meinenhofer, Academic Press, New York, 1981, which is incorporated herein by reference in its entirety. For example, the protecting group removing agent is selected from the group consisting of Pd / K2CO3, 1,3-dimethylbarbituric acid / Pd(PPh3)4, Pd(PPh)3, PhSiH3, H2, piperidine, and trifluoroacetic acid (TFA).

[0407] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0408] [ka] providing a compound of formula (XI) having the formula: forming a compound of formula (X) from a compound of formula (XI) may include:

[0409] In one embodiment, the compound of formula (XI) has the structure:

[0410] [ka]

[0411] In some embodiments, the process for forming a compound of formula (X) comprises: The compound of formula (XI) may be reacted with a compound of formula (XII):

[0412] [ka] or a salt thereof, Producing a compound of formula (X) Includes:

[0413] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0414] [ka] providing a compound of formula (XIII) having the formula: forming a compound of formula (XI) from a compound of formula (XIII) may include:

[0415] In one embodiment, the compound of formula (XIII) has the structure:

[0416] [ka]

[0417] In some embodiments, the process of forming a compound of formula (XI) comprises reacting a compound of formula (XIII) with an oxidizing agent to produce a compound of formula (XI).

[0418] According to the present disclosure, any suitable oxidizing agent capable of converting a primary alcohol into an aldehyde group can be used. Suitable oxidizing agents that can be used are known to those skilled in the art. For example, the oxidizing agent can be selected from the group consisting of pyridinium chlorochromate (PCC), pyridinium dichromate (PDC), silver carbonate (AgCO), tetra-n-propylammonium perruthenate (TPAP), and Dess-Martin periodinane (DMP).

[0419] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0420] [ka] [In the formula, Y is -OC 1~6 wherein R is an alkyl group; and forming a compound of formula (XIII) from a compound of formula (XIV); may include:

[0421] In one embodiment, the compound of formula (XIV) has the structure:

[0422] [ka]

[0423] In some embodiments, the process of forming a compound of Formula (XIII) comprises reacting a compound of Formula (XIV) with a reducing agent.

[0424] According to the present disclosure, any suitable reducing agent capable of converting an ester into a primary alcohol group can be used. Suitable reducing agents that can be used are known to those skilled in the art. For example, the reducing agent is selected from the group consisting of LiAlH4, DIBALH, and LiBH4.

[0425] In some embodiments, the process for preparing a compound of formula (I) comprises: structure:

[0426] [ka] providing a compound of formula (XV) having the formula: forming a compound of formula (XIV) from a compound of formula (XV) may include:

[0427] In one embodiment, the compound of formula (XV) has the structure:

[0428] [ka]

[0429] In some embodiments, the process of forming a compound of formula (XIV) comprises reacting a compound of formula (XV) with a protecting group introducing agent to produce a compound of formula (XIV).

[0430] According to the present disclosure, any suitable protecting group introducing agent can be used to protect the hydroxyl group of the compound of formula (XV). Suitable protecting group introducing agents that can be used are known to those skilled in the art. In some embodiments, the protecting group introducing agent is selected from the group consisting of TBSCl, TBSOTf, TMSCl, TMSOTf, TESCl, TESOTf, TBDPSCl, TBDPSOTf, TIPSCl, and TIPSOTf.

[0431] In another aspect, the present disclosure provides a product prepared according to any of the methods described in this disclosure.

[0432] In one embodiment, the product has formula (I):

[0433] [ka] [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R 1 is C 1~5 is alkyl, R 2 is, independently at each occurrence, optionally replaced by C 10~30 is alkyl, R 3 is, independently at each occurrence, optionally replaced by C 10~30 alkyl] or a salt thereof.

[0434] In another embodiment, the product has formula (Ia):

[0435] [ka] or a salt thereof.

[0436] In another aspect, the present disclosure provides a compound of formula (I):

[0437] [ka] [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R 1 is C 1~5 is alkyl, R 2 is, independently at each occurrence, optionally replaced by C 10~30 is alkyl, R 3 is, independently at each occurrence, optionally replaced by C 10~30 alkyl] or a salt thereof, Methods for making peptides or peptidomimetics are provided.

[0438] In at least one embodiment, the compounds of formula (I) are used as supports or tags for generating peptides or peptidomimetics.

[0439] In one embodiment, the peptidomimetic has the formula:

[0440] [ka] or a salt thereof.

[0441] In one embodiment, the peptidomimetic has the formula:

[0442] [ka] or a salt thereof.

[0443] In another aspect, the present disclosure provides a compound of formula (I):

[0444] [ka] [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R 1 is C 1~5 is alkyl, R 2 is optionally replaced by C for each occurrence. 10~30 independently selected from alkyl, R 3 is optionally replaced by C for each occurrence. 10~30 alkyl] or a salt thereof.

[0445] In one embodiment, the compound has the formula:

[0446] [ka] or a salt thereof.

[0447] In one embodiment, the compound has the formula:

[0448] [ka] or a salt thereof.

[0449] The present disclosure provides compounds of formula (AI):

[0450] [ka] [In the formula, m and n are independently an integer of 0 to 5; R 1a is C 1~5 is alkyl, R 2a is, independently at each occurrence, optionally replaced by C 10~30 is alkyl, R 3a is, independently at each occurrence, optionally replaced by C 10~30 alkyl] The present invention provides a compound having the structure:

[0451] According to the present disclosure, m may be an integer from 0 to 5, and n may be an integer from 0 to 5. For example, m may be 1, 2, 3, 4, or 5, and n may be 1, 2, 3, 4, or 5.

[0452] According to this disclosure, R 1a can be methyl, ethyl, propyl, butyl, or pentyl. For example, R 1 can be n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, or n-pentyl. 1a is methyl.

[0453] According to this disclosure, R 2a is, independently at each occurrence, optionally replaced by C 10~30 It is an alkyl group. For example, C 10~30 Alkyl may be optionally substituted 1 to 10 times with substituents selected from halogen, hydroxy, lower alkoxy, carboxy, carboalkoxy (also called alkoxycarbonyl), carboxamido (also called alkylaminocarbonyl), cyano, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, phenyl, benzyl, heteroaryl, phenoxy, or benzyloxy. For example, C 10~30The alkyl may be optionally substituted 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times with any of the above substituents, each substituent independently selected from halogen, hydroxy, lower alkoxy, carboxy, carboalkoxy, carboxamido, cyano, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, phenyl, benzyl, heteroaryl, phenoxy, or benzyloxy.

[0454] According to this disclosure, R 3a is, independently at each occurrence, optionally replaced by C 10~30 It is an alkyl group. For example, C 10~30 Alkyl may be optionally substituted 1 to 10 times with substituents selected from halogen, hydroxy, lower alkoxy, carboxy, carboalkoxy (also called alkoxycarbonyl), carboxamido (also called alkylaminocarbonyl), cyano, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, phenyl, benzyl, heteroaryl, phenoxy, or benzyloxy. For example, C 10~30 The alkyl may be optionally substituted 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times with any of the above substituents, each substituent independently selected from halogen, hydroxy, lower alkoxy, carboxy, carboalkoxy, carboxamido, cyano, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, phenyl, benzyl, heteroaryl, phenoxy, or benzyloxy.

[0455] In at least one embodiment, R 2a is, independently at each occurrence, optionally replaced by C 18~26 alkyl, and R 3a is, independently at each occurrence, optionally replaced by C 18~26 For example, each R 2a are independently optionally substituted C 22 alkyl, and each R 3a are independently optionally substituted C22 It is alkyl.

[0456] The present disclosure provides a compound of formula (BI):

[0457] [ka] [In the formula, m is an integer from 1 to 5; R 1b is C 1~5 is alkyl, R 2b is, independently at each occurrence, optionally replaced by C 10~30 alkyl] The present invention provides a compound having the structure:

[0458] According to the present disclosure, m may be an integer from 1 to 5. For example, m may be 1, 2, 3, 4, or 5.

[0459] According to this disclosure, R 1b can be methyl, ethyl, propyl, butyl, or pentyl. For example, R 1b can be n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, or n-pentyl. 1b is methyl.

[0460] According to this disclosure, R 2b is, independently at each occurrence, optionally replaced by C 10~30 It is an alkyl group. For example, C 10~30 Alkyl may be optionally substituted 1 to 10 times with substituents selected from halogen, hydroxy, lower alkoxy, carboxy, carboalkoxy (also called alkoxycarbonyl), carboxamido (also called alkylaminocarbonyl), cyano, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, phenyl, benzyl, heteroaryl, phenoxy, or benzyloxy. For example, C 10~30The alkyl may be optionally substituted 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times with any of the above substituents, each substituent independently selected from halogen, hydroxy, lower alkoxy, carboxy, carboalkoxy, carboxamido, cyano, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, phenyl, benzyl, heteroaryl, phenoxy, or benzyloxy.

[0461] In at least one embodiment, R 2b is, independently at each occurrence, optionally replaced by C 18~26 For example, each R 2b are independently optionally substituted C 22 It is alkyl.

[0462] The present disclosure provides compounds of formula (CI):

[0463] [ka] [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R 2c is, independently at each occurrence, optionally replaced by C 10~30 is alkyl, R 3c is, independently at each occurrence, optionally replaced by C 10~30 alkyl] The present invention provides a compound having the structure:

[0464] According to the present disclosure, m can be an integer from 1 to 5, and n can be an integer from 1 to 5, provided that (m+n) > 3. For example, m can be 1, 2, 3, 4, or 5, and n can be 1, 2, 3, 4, or 5.

[0465] According to this disclosure, R 2c is, independently at each occurrence, optionally replaced by C 10~30It is an alkyl group. For example, C 10~30 Alkyl may be optionally substituted 1 to 10 times with substituents selected from halogen, hydroxy, lower alkoxy, carboxy, carboalkoxy (also called alkoxycarbonyl), carboxamido (also called alkylaminocarbonyl), cyano, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, phenyl, benzyl, heteroaryl, phenoxy, or benzyloxy. For example, C 10~30 The alkyl may be optionally substituted 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times with any of the above substituents, each substituent independently selected from halogen, hydroxy, lower alkoxy, carboxy, carboalkoxy, carboxamido, cyano, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, phenyl, benzyl, heteroaryl, phenoxy, or benzyloxy.

[0466] According to this disclosure, R 3c is, independently at each occurrence, optionally replaced by C 10~30 It is an alkyl group. For example, C 10~30 Alkyl may be optionally substituted 1 to 10 times with substituents selected from halogen, hydroxy, lower alkoxy, carboxy, carboalkoxy (also called alkoxycarbonyl), carboxamido (also called alkylaminocarbonyl), cyano, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, phenyl, benzyl, heteroaryl, phenoxy, or benzyloxy. For example, C 10~30 The alkyl may be optionally substituted 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times with any of the above substituents, each substituent independently selected from halogen, hydroxy, lower alkoxy, carboxy, carboalkoxy, carboxamido, cyano, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, phenyl, benzyl, heteroaryl, phenoxy, or benzyloxy.

[0467] In at least one embodiment, R 2c is, independently at each occurrence, optionally replaced by C 18~26 alkyl, and R 3c is, independently at each occurrence, optionally replaced by C 18~26 For example, each R 2c are independently optionally substituted C 22 alkyl, and each R 3c are independently optionally substituted C 22 It is alkyl.

[0468] Methods for producing peptides or peptidomimetics using tags The present disclosure provides a method of making a peptide or peptidomimetic, comprising: (a) Formula (I):

[0469] [ka] [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R 1 is C 1~5 is alkyl, R 2 is optionally replaced by C for each occurrence. 10~30 independently selected from alkyl, R 3 is optionally replaced by C for each occurrence. 10~30 alkyl] or a salt thereof, (b) providing a first amino acid (AA1), and (c) coupling a first amino acid (AA1) with the compound of formula (I) to form a peptide bond between the first amino acid (AA1) and the compound of formula (I); The present invention provides a method comprising:

[0470] The term "amino acid" refers to a molecule containing both an amino group and a carboxyl group. Amino acids include alpha-amino acids and beta-amino acids, the structures of which are depicted below. In certain embodiments, the amino acid is an alpha-amino acid.

[0471] [ka]

[0472] Suitable amino acids that may be used in accordance with the present disclosure include, but are not limited to, i) natural alpha-amino acids, such as the 20 common naturally occurring alpha-amino acids found in peptides and proteins: alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, and the D and L isomers of their derivatives; ii) non-natural alpha-amino acids; iii) natural beta-amino acids (e.g., beta-alanine); and iv) non-natural beta-amino acids.

[0473] In one embodiment, step (c) of coupling AA1 with a compound of formula (I) comprises coupling the carboxylic acid moiety of AA1 with the amino moiety of the compound of formula (I).

[0474] In some embodiments, the first amino acid (AA1) contains a protected amino group.

[0475] In one embodiment, the first amino acid (AA1) is:

[0476] [ka]

[0477] In some embodiments, the method of making a peptide or peptidomimetic may include activating a chemical group on the first amino acid (AA1) to prepare the first amino acid (AA1) for coupling with a compound of formula (I) prior to step (c).

[0478] In some embodiments, the method of making a peptide or peptidomimetic comprises: (d) removing the protecting group from the first amino acid (AA1); (e) providing a second amino acid (AA2), and (f) coupling a second amino acid (AA2) with the first amino acid (I-AA1) to form a peptide bond (I-AA1-AA2) between the second amino acid (AA2) and the first amino acid; may include:

[0479] In one embodiment, step (f) comprises coupling the carboxylic acid moiety of AA2 with the amino moiety of AA1.

[0480] In one embodiment, the second amino acid (AA2) is:

[0481] [ka]

[0482] In some embodiments, the method of making a peptide or peptidomimetic comprises: (g) removing the protecting group from the second amino acid (AA2); (h) providing a third amino acid (AA3), and (i) coupling a third amino acid (AA3) with a second amino acid to form a peptide bond (I-AA1-AA2-AA3) between the third amino acid (AA3) and the second amino acid; may include:

[0483] In one embodiment, step (i) comprises coupling the carboxylic acid moiety of AA3 with the amino moiety of AA2.

[0484] In one embodiment, the third amino acid (AA3) is:

[0485] [ka]

[0486] In some embodiments, the method of making a peptide or peptidomimetic comprises: (j) removing the protecting group from the third amino acid (AA3); (k) providing a fourth amino acid (AA4), and (l) coupling a fourth amino acid (AA4) with a third amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4) between the fourth amino acid (AA4) and the third amino acid; may include:

[0487] In one embodiment, step (l) comprises coupling the carboxylic acid moiety of AA4 with the amino moiety of AA3.

[0488] In one embodiment, the fourth amino acid (AA4) is:

[0489] [ka]

[0490] In some embodiments, the method of making a peptide or peptidomimetic comprises: (m) removing the protecting group from the fourth amino acid (AA4); (n) providing a fifth amino acid (AA5), and (o) coupling a fifth amino acid (AA5) with a fourth amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5) between the fifth amino acid (AA5) and the fourth amino acid; may include:

[0491] In one embodiment, step (o) comprises coupling the carboxylic acid moiety of AA5 with the amino moiety of AA4.

[0492] In one embodiment, the fifth amino acid (AA5) is:

[0493] [ka]

[0494] In some embodiments, the method of making a peptide or peptidomimetic comprises: (p) removing the protecting group from the fifth amino acid (AA5); (q) providing a sixth amino acid (AA6), and (r) coupling the sixth amino acid (AA6) with the fifth amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5-AA6) between the sixth amino acid (AA6) and the fifth amino acid; may include:

[0495] In one embodiment, step (r) comprises coupling the carboxylic acid moiety of AA6 with the amino moiety of AA5.

[0496] In one embodiment, the sixth amino acid (AA6) is:

[0497] [ka]

[0498] In some embodiments, the method of making a peptide or peptidomimetic comprises: (s) removing the protecting group from the sixth amino acid (AA6); (t) providing a seventh amino acid (AA7), and (u) coupling the seventh amino acid (AA6) with the sixth amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5-AA6-AA7) between the seventh amino acid (AA7) and the sixth amino acid; may include:

[0499] In one embodiment, step (u) comprises coupling the carboxylic acid moiety of AA7 with the amino moiety of AA6.

[0500] In one embodiment, the seventh amino acid (AA7) is:

[0501] [ka]

[0502] In some embodiments, the method of making a peptide or peptidomimetic comprises: (v) removing the protecting group from the seventh amino acid (AA7); (w) providing an eighth amino acid (AA8), and (x) coupling the eighth amino acid (AA8) with the seventh amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5-AA6-AA7-AA8) between the eighth amino acid (AA8) and the seventh amino acid; may include:

[0503] In one embodiment, step (x) comprises coupling the carboxylic acid moiety of AA8 with the amino moiety of AA7.

[0504] In one embodiment, the eighth amino acid (AA8) is:

[0505] [ka]

[0506] In some embodiments, the method of making a peptide or peptidomimetic comprises: (y) removing the protecting group from the eighth amino acid (AA8); (z) providing a ninth amino acid (AA9), and (aa) coupling the ninth amino acid (AA9) with the eighth amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5-AA6-AA7-AA8-AA9) between the ninth amino acid (AA9) and the eighth amino acid; may include:

[0507] In one embodiment, step (aa) comprises coupling the carboxylic acid moiety of AA9 with the amino moiety of AA8.

[0508] In one embodiment, the ninth amino acid (AA9) is:

[0509] [ka]

[0510] In some embodiments, the method of making a peptide or peptidomimetic comprises: (bb) removing the protecting group from the ninth amino acid (AA9); (cc) providing a tenth amino acid (AA10), and (dd) coupling the tenth amino acid (AA10) with the ninth amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5-AA6-AA7-AA8-AA9-AA10) between the tenth amino acid (AA10) and the ninth amino acid; may include:

[0511] In one embodiment, step (dd) comprises coupling the carboxylic acid moiety of AA10 with the amino moiety of AA9.

[0512] In one embodiment, the tenth amino acid (AA10) is:

[0513] [ka]

[0514] In some embodiments, the method of making a peptide or peptidomimetic comprises: (i) Amino acids (AA n removing the protecting group from (ii) the subsequent amino acid (AA n+1 ), and (iii) the subsequent amino acid (AA n+1 ) to amino acids (AA n ) to form the next amino acid (AA n+1 ) and amino acids (AA n forming a peptide bond between and repeating The repeating step may be performed 1 to 100 times.

[0515] In some embodiments, the method of making a peptide or peptidomimetic comprises: (ee) removing the protecting group from the tenth amino acid (AA10); (ff) Formula (B):

[0516] [ka] wherein x is an integer from 1 to 15; and (gg) coupling the compound of formula (B) with one of the amino acids in the peptide or peptidomimetic to form a triazole ring. may include:

[0517] In one embodiment, the compound of formula (B) has the formula:

[0518] [ka]

[0519] In some embodiments of the present disclosure, one or more peptides are n For example, in some embodiments, a method of making a peptide or peptidomimetic comprises: (i) Amino acids (AA n removing the protecting group from (ii) Peptide (AA m1 -AA mp ) [where p is between 2 and 20], and (iii) Peptide (AA m -AA mp ) amino acid (AA n ) to form peptide (AA m1 -AA mp ) and amino acids (AA n forming a peptide bond between and repeating The repeating step may be performed 1 to 100 times.

[0520] According to the present disclosure, peptide (AA m1 -AA mp ), m may be 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. In some embodiments, m is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In some embodiments, m is 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0521] According to the present disclosure, peptide (AA m1 -AA mp) may be a dipeptide, tripeptide, tetrapeptide, pentapeptide, hexapeptide, heptapeptide, octapeptide, nonapeptide, or decapeptide.

[0522] Peptides may contain all D-amino acids, all L-amino acids, or a mixture of L- and D-amino acids. In preferred embodiments, peptides may contain only D-amino acids or a mixture of D- and L-amino acids, with a D-amino acid content of greater than 50%, 60%, 70%, 80%, 90%, or 95%.

[0523] During synthesis, non-participating carboxylic acids or amines for a reactive set of amino acids or peptide fragments may be protected with a suitable protecting group (PG) that can be later selectively removed if desired. A detailed description of these groups, as well as their selection and chemistry, is contained in "The Peptides, Vol. 3," edited by Gross and Meinenhofer, Academic Press, New York, 1981, which is incorporated herein by reference in its entirety.

[0524] In some embodiments, the method of making a peptide or peptidomimetic according to any of the above embodiments comprises: (hh) further cleaving the peptide bond between the first amino acid (AA1) and the compound of formula (I) to obtain a peptide or peptidomimetic.

[0525] According to the present disclosure, the first amino acid (AA1), the second amino acid (AA2), the third amino acid (AA3), the fourth amino acid (AA4), the fifth amino acid (AA5), the sixth amino acid (AA6), the seventh amino acid (AA7), the eighth amino acid (AA8), the ninth amino acid (AA9), the tenth amino acid (AA10), and any subsequent amino acids (AA n+1 ) are, respectively,

[0526] [ka] [In the formula, PG 5 is a suitable protecting group.

[0527] According to the present disclosure, any suitable protecting group that can be used to protect the amino group of an amino acid is PG 5 Suitable protecting groups that can be used are known to those skilled in the art. A detailed description of these groups, as well as their selection and chemistry, is contained in "The Peptides, Vol. 3," edited by Gross and Meinenhofer, Academic Press, New York, 1981, which is incorporated herein by reference in its entirety. For example, PG 5 Each occurrence may be independently selected from the group consisting of triphenylmethyl (Trt), allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

[0528] According to the present disclosure, the first amino acid (AA1), the second amino acid (AA2), the third amino acid (AA3), the fourth amino acid (AA4), the fifth amino acid (AA5), the sixth amino acid (AA6), the seventh amino acid (AA7), the eighth amino acid (AA8), the ninth amino acid (AA9), the tenth amino acid (AA10), and any subsequent amino acids (AA n+1 ) are, respectively,

[0529] [ka] may be independently selected from the group consisting of:

[0530] In one embodiment, the peptidomimetic has the formula:

[0531] [ka] or a salt thereof.

[0532] In one embodiment, the peptidomimetic has the formula:

[0533] [ka] or a salt thereof.

[0534] According to the present disclosure, the amino acids that can be used to prepare peptides and peptidomimetics can be both natural and unnatural amino acids.

[0535] The present disclosure provides a compound of formula (DI):

[0536] [ka] [In the formula, k is an integer from 1 to 15, q is an integer from 0 to 10, R 1d is C 1~6 is alkyl, PG 6 is a suitable protecting group, PG 7 is a suitable protecting group] or a salt thereof, the method comprising: (a) Structure:

[0537] [ka] providing a compound of formula (DII) having the formula: (b) forming a compound of formula (DI) from a compound of formula (DII); Includes:

[0538] According to this disclosure, PG 6is any suitable protecting group that can be used to protect an amino group. Suitable protecting groups that can be used are known to those skilled in the art. A detailed description of these groups, as well as their selection and chemistry, is contained in "The Peptides, Vol. 3," edited by Gross and Meinenhofer, Academic Press, New York, 1981, which is incorporated herein by reference in its entirety. For example, PG 6 may be selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc). 6 is 9-fluorenylmethoxycarbonyl (Fmoc).

[0539] According to this disclosure, PG 7 is any suitable protecting group that can be used to protect an amino group. Suitable protecting groups that can be used are known to those skilled in the art. A detailed description of these groups, as well as their selection and chemistry, is contained in "The Peptides, Vol. 3," edited by Gross and Meinenhofer, Academic Press, New York, 1981, which is incorporated herein by reference in its entirety. For example, PG 7 may be selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc). 7 is tert-butyloxycarbonyl (Boc).

[0540] In one embodiment, the compound of formula (DI) has the structure of formula (DIa) or formula (DIb).

[0541] [ka]

[0542] In another embodiment, the compound of formula (DII) has the structure of formula (DIIa) or formula (DIIb).

[0543] [ka]

[0544] In some embodiments, step (b) of forming a compound of formula (DI) comprises: The compound of formula (DII) may be reacted with a compound of formula (DIII):

[0545] [ka] with a compound of formula (II) Includes:

[0546] In one embodiment, the compound of Formula (DIII) has the structure of Formula (DIIIa):

[0547] [ka]

[0548] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0549] [ka] or a salt thereof; and Before step (a), forming a compound of formula (DII) from a compound of formula (DIV). may include:

[0550] In one embodiment, the compound of formula (DIV) has the structure of formula (DIVa) or formula (DIVb).

[0551] [ka]

[0552] In some embodiments, the step of forming a compound of formula (DII) comprises: The compound of formula (DIV) is reacted with a compound of formula (DVa) PG 6 -X(DVa) or Formula (DVb) PG 6 2O(DVb) [In the formula, X is OSu, OTf, Cl, or Br, Producing a compound of formula (DII) Includes:

[0553] According to the present disclosure, the compound of formula (DVa) or formula (DVb) may be selected from the group consisting of AllocCl, Alloc2O, Cbz2O, CbzCl, FmocCl, FmocOSu, and Boc2O.

[0554] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0555] [ka] providing a compound of formula (DVI) having the formula: forming a compound of formula (DIV) from a compound of formula (DVI) may include:

[0556] In some embodiments, the step of forming a compound of formula (DIV) comprises: reacting a compound of formula (DVI) with a deprotecting agent to form a compound of formula (DIV). Includes:

[0557] According to the present disclosure, any suitable deprotecting agent can be used to remove the protecting group PG from the amino group of the compound of formula (DVI). 8Suitable protecting group removing agents that can be used to remove specific protecting groups are known to those skilled in the art. A detailed description of these groups, as well as their selection and chemistry, is contained in "The Peptides, Vol. 3," edited by Gross and Meinenhofer, Academic Press, New York, 1981, which is incorporated herein by reference in its entirety. For example, the protecting group removing agent can be selected from the group consisting of Pd(PPh)3, PhSiH3, H2, HCl, piperidine, and trifluoroacetic acid (TFA).

[0558] In one embodiment, the compound of Formula (DVI) has the structure of Formula (DVIa) or Formula (DVIb):

[0559] [ka]

[0560] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0561] [ka] providing a compound of formula (DVII) having the formula: forming a compound of formula (DVI) from a compound of formula (DVII) may include:

[0562] In some embodiments, the step of forming a compound of Formula (DVI) comprises: reacting a compound of formula (DVII) with a base to form a compound of formula (DVI) Includes:

[0563] According to the present disclosure, any suitable base can be used to hydrolyze the methyl ester group in the compound of formula (DVII). In some embodiments, the base is selected from the group consisting of LiOH, NaOH, and KOH.

[0564] In one embodiment, the compound of formula (DVII) has the structure of formula (DVIIa) or formula (DVIIb):

[0565] [ka]

[0566] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0567] [ka] [In the formula, and LG′ is a suitable living group; and forming a compound of formula (DVII) from a compound of formula (DVIII) may include:

[0568] In some embodiments, the step of forming a compound of Formula (DVII) comprises: reacting a compound of formula (DVIII) with an azide group introducing agent to produce a compound of formula (DVII); Includes:

[0569] Azide group-introducing agents that can be used to implement the above embodiment are known to those skilled in the art. In one embodiment, the azide group-introducing agent is NaN3, TMSN3, (PhO)2P(O)N3, Zn(N3)2 * 2Py, and n-Bu4NN3.

[0570] In one embodiment, LG' is selected from the group consisting of halogen, OTf, OMs, and OTs.

[0571] In another embodiment, the compound of formula (DVIII) has the structure of formula (DVIIIa) or formula (DVIIIb).

[0572] [ka]

[0573] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0574] [ka] [In the formula, wherein LG″ is a suitable living group; and forming a compound of formula (DVIII) from a compound of formula (DIX) may include:

[0575] In one embodiment, LG″ is selected from the group consisting of halogen, OTf, OMs, and OTs.

[0576] In some embodiments, the step of forming a compound of formula (DVIII) comprises: The compound of formula (DIX) may be reacted with a compound of formula (DX):

[0577] [ka] [In the formula, PG 8 is a suitable protecting group, Producing a compound of formula (DVII) Includes:

[0578] According to this disclosure, PG 8is any suitable protecting group that can be used to protect an amino group. Suitable protecting groups that can be used are known to those skilled in the art. A detailed description of these groups, as well as their selection and chemistry, is contained in "The Peptides, Vol. 3," edited by Gross and Meinenhofer, Academic Press, New York, 1981, which is incorporated herein by reference in its entirety. For example, PG 8 may be selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc). 8 is tert-butyloxycarbonyl (Boc).

[0579] In another embodiment, the compound of Formula (DIX) has the structure of Formula (DIXa):

[0580] [ka]

[0581] In yet another embodiment, the compound of formula (DX) has the structure of formula (DXa).

[0582] [ka]

[0583] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0584] [ka] [In the formula, R 2d is C 1~6 wherein R is an alkylene; and forming a compound of formula (DIX) from a compound of formula (DXI) may include:

[0585] In some embodiments, the step of forming a compound of formula (DIX) comprises reacting a compound of formula (DXI) with a reducing agent to produce a compound of formula (DIX).

[0586] According to the present disclosure, any reducing agent capable of reducing an alkene double bond can be used to prepare a compound of formula (DIX). In some embodiments, the reducing agent is selected from the group consisting of H, NHHCO, NABH(OAc), and LiAlH.

[0587] In one embodiment, the compound of formula (DXI) has the structure of formula (DXIa):

[0588] [ka]

[0589] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0590] [ka] providing a compound of formula (DXII) having the formula: forming a compound of formula (DXI) from a compound of formula (DXII) may include:

[0591] In some embodiments, the step of forming a compound of formula (DXI) comprises: The compound of formula (DXII) may be reacted with a compound of formula (DXIII): PPh3 * C 1~6 Alkyl-Hal(DXIII) [In the formula, where Hal is a halogen, Producing a compound of formula (DXI) Includes:

[0592] In one embodiment, the compound of formula (DXII) has the structure of formula (DXIIa):

[0593] [ka]

[0594] In another embodiment, the compound of formula (DXIII) has the structure of formula (DXIIIa): PPh3 * MeBr(DXIIIa)

[0595] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0596] [ka] providing a compound of formula (DXIV) having the formula: forming a compound of formula (DXII) from a compound of formula (DXIV) may include:

[0597] In some embodiments, the step of forming a compound of formula (DXII) comprises: The compound of formula (DXIV) may be reacted with a compound of formula (DXV):

[0598] [ka] [In the formula, wherein LG"' is a suitable living group, Producing a compound of formula (DXII) Includes:

[0599] In one embodiment, LG"' is selected from the group consisting of halogen, OTf, OMs, and OTs.

[0600] In another embodiment, the compound of formula (DXIV) has the structure of formula (DXIVa).

[0601] [ka]

[0602] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0603] [ka] [In the formula, LG * is a suitable living group; and forming a compound of formula (DX) from a compound of formula (DXVI) may include:

[0604] In one embodiment, LG * is selected from the group consisting of halogens, OTf, OMs, and OTs.

[0605] In some embodiments, the step of forming a compound of formula (DX) comprises: The compound of formula (DXVI) may be reacted with a compound of formula (DXVII):

[0606] [ka] with a compound of formula (DX) Includes.

[0607] In one embodiment, the compound of formula (DXVI) has the structure of formula (DXVIc):

[0608] [ka]

[0609] In some embodiments, the process for preparing a compound of formula (DI) comprises: structure:

[0610] [ka] providing a compound of formula (DXVIII) having the formula: forming a compound of formula (DXVI) from a compound of formula (DXVIII) may include:

[0611] In some embodiments, the step of forming a compound of Formula (DXVI) comprises: The compound of formula (DXVIII) is reacted with a compound of formula (DXIXa) LG * -X(DXIXa) or Formula (DXIXb) LG * 2O(DXIXb) [In the formula, X is OSu, Cl, or Br, Producing a compound of formula (DXVI) Includes:

[0612] In one embodiment, the compound of Formula (DXIXa) or Formula (DXIXb) is selected from the group consisting of TfCl, Tf2O, MsCl, and Ms2O.

[0613] The present disclosure provides a compound of formula (DII):

[0614] [ka] [In the formula, q is an integer from 0 to 10, R 1d is C 1~6 is alkyl, PG 6 is a suitable protecting group] or a salt thereof, said process comprising: (a) Structure:

[0615] [ka] or a salt thereof; and (b) forming a compound of formula (DII) from a compound of formula (DXXI); The present invention provides a method comprising:

[0616] In some embodiments, PG 6 is selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

[0617] In some embodiments, the compound of formula (DXXI) is treated with the following compound:

[0618] [ka]

[0619] In some embodiments, the compound of Formula (DXXI) is treated with a catalyst. In some embodiments, the catalyst is Pd(dppf)Cl-DCM.

[0620] In some embodiments, the compound of Formula (DXXI) is treated with a base. In some embodiments, the base is K2CO3.

[0621] In some embodiments, the preparation is carried out in a solvent mixture composed of H2O / iPrOH.

[0622] In some embodiments, the compound of Formula (DII) has the structure of Formula (DIIa) or Formula (DIIb).

[0623] [ka]

[0624] In some embodiments, the compound of formula (DXXI) is

[0625] [ka] of

[0626] [ka] It is formed by exposure to

[0627] In some embodiments, the compound of Formula (DXXI) is formed in the presence of PPh3.

[0628] In some embodiments, the compound of Formula (DXXI) is formed in the presence of DIAD.

[0629] In some embodiments, the compound of formula (DXXI) is formed in the presence of 4 Å molecular sieves.

[0630] In some embodiments, the compound of Formula (DXXI) is formed in an organic solvent. In some embodiments, the organic solvent is THF.

[0631] In some embodiments,

[0632] [ka] teeth,

[0633] [ka] is formed by exposing it to PBin2.

[0634] In some embodiments,

[0635] [ka] is formed in the presence of Pd(OAc)2.

[0636] In some embodiments

[0637] [ka] is formed in the presence of PCy3.

[0638] In some embodiments,

[0639] [ka] is formed in the presence of KOAc.

[0640] In some embodiments,

[0641] [ka] is formed in the presence of dioxane.

[0642] In some embodiments,

[0643] [ka] is formed at elevated temperatures over a period of time.

[0644] In some embodiments,

[0645] [ka] teeth,

[0646] [ka] Bu4N + Br3 -It is formed by exposure to

[0647] In some embodiments,

[0648] [ka] is formed in an organic solvent. In some embodiments, the organic solvent is a mixture of DCM and methanol.

[0649] The present disclosure provides a compound of formula (DII):

[0650] [ka] [In the formula, q is an integer from 0 to 10, R 1d is C 1~6 is alkyl, PG 6 is a suitable protecting group] or a salt thereof, said process comprising: (a) Structure:

[0651] [ka] or a salt thereof; and (b) forming a compound of formula (DII) from a compound of formula (DIV); The present invention provides a process including:

[0652] According to this disclosure, PG 6 is any suitable protecting group that can be used to protect an amino group. Suitable protecting groups that can be used are known to those skilled in the art. A detailed description of these groups, as well as their selection and chemistry, is contained in "The Peptides, Vol. 3," edited by Gross and Meinenhofer, Academic Press, New York, 1981, which is incorporated herein by reference in its entirety. For example, PG 6may be selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc). 6 is 9-fluorenylmethoxycarbonyl (Fmoc).

[0653] In one embodiment, the compound of formula (DII) has the structure of formula (DIIa) or formula (DIIb):

[0654] [ka]

[0655] In another embodiment, the compound of formula (DIV) has the structure of formula (DIVa) or formula (DIVb).

[0656] [ka]

[0657] In some embodiments, the process for forming the compound of Formula (DII) comprises: The compound of formula (DIV) is reacted with a compound of formula (DVa) PG 6 -X(DVa) or Formula (DVb) PG 6 2O(DVb) [In the formula, X is OSu, OTf, Cl, or Br, Producing a compound of formula (DII) Includes:

[0658] According to the present disclosure, the compound of formula (DVa) or formula (DVb) is selected from the group consisting of AllocCl, Alloc2O, Cbz2O, CbzCl, FmocCl, FmocOSu, and Boc2O.

[0659] In some embodiments, the process for preparing a compound of formula (DII) comprises: structure:

[0660] [ka] [In the formula, PG 8 is a suitable protecting group; and forming a compound of formula (DIV) from a compound of formula (DXVI) may include:

[0661] According to this disclosure, PG 8 is any suitable protecting group that can be used to protect an amino group. Suitable protecting groups that can be used are known to those skilled in the art. A detailed description of these groups, as well as their selection and chemistry, is contained in "The Peptides, Vol. 3," edited by Gross and Meinenhofer, Academic Press, New York, 1981, which is incorporated herein by reference in its entirety. For example, PG 8 may be selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc). 8 is tert-butyloxycarbonyl (Boc).

[0662] In some embodiments, the step of forming a compound of formula (DIV) comprises reacting a compound of formula (DXVI) with a deprotecting agent to produce a compound of formula (DIV).

[0663] According to the present disclosure, any suitable deprotecting agent can be used to remove the protecting group PG from the amino group of the compound of formula (DVI). 8

[0033] Suitable protecting group removing agents that can be used to remove particular protecting groups are known to those skilled in the art. A detailed description of these groups, as well as their selection and chemistry, is contained in "The Peptides, Vol. 3," edited by Gross and Meinenhofer, Academic Press, New York, 1981, which is incorporated herein by reference in its entirety. In some embodiments, the protecting group removing agent is selected from the group consisting of HCl, Pd(PPh)3, PhSiH3, H2, piperidine, and trifluoroacetic acid (TFA).

[0664] In one embodiment, the compound of formula (DXVI) has the structure of formula (DXVIa) or formula (DXVIb).

[0665] [ka]

[0666] In some embodiments, the process for preparing a compound of formula (DII) comprises: structure:

[0667] [ka] [In the formula, and LG′ is a suitable living group; and forming a compound of formula (DXVI) from a compound of formula (DXVII) may include:

[0668] In some embodiments, the step of forming a compound of formula (DXVI) comprises reacting a compound of formula (DXVII) with an azide group introducing agent to produce a compound of formula (DXVI).

[0669] In some embodiments, the azide group introducing agent is NaN3, TMSN3, (PhO)2P(O)N3, Zn(N3)2 *2Py, and n-Bu4NN3.

[0670] In some embodiments, LG' is selected from the group consisting of halogen, OTf, OMs, and OTs.

[0671] In one embodiment, the compound of formula (DXVII) has the structure of formula (DXVIIa) or formula (DXVIIb).

[0672] [ka]

[0673] In some embodiments, the process for preparing a compound of formula (DII) comprises: structure:

[0674] [ka] [In the formula, wherein LG″ is a suitable living group; and forming a compound of formula (DXVII) from a compound of formula (DIX) may include:

[0675] In some embodiments, LG″ is selected from the group consisting of halogen, OTf, OMs, and OTs.

[0676] In some embodiments, the step of forming a compound of formula (DXVII) comprises: The compound of formula (DIX) may be reacted with a compound of formula (DXVIII):

[0677] [ka] with a compound of formula (DXVII) Includes:

[0678] In one embodiment, the compound of formula (DIX) has the structure of formula (DIXa):

[0679] [ka]

[0680] In another embodiment, the compound of formula (DXVIII) has the structure of formula (DXVIIIa).

[0681] [ka]

[0682] In some embodiments, the process for preparing a compound of formula (DII) comprises: structure:

[0683] [ka] providing a compound of formula (DXIX) having the formula: forming a compound of formula (DIX) from a compound of formula (DXIX) may include:

[0684] In some embodiments, The step of forming a compound of formula (DIX) comprises: The compound of formula (DXIX) may be reacted with a compound of formula (DXV):

[0685] [ka] [In the formula, wherein LG"' is a suitable living group, Producing a compound of formula (DXIX) Includes:

[0686] In one embodiment, LG"' is selected from the group consisting of halogen, OTf, OMs, and OTs.

[0687] In another embodiment, the compound of formula (DXIX) has the structure of formula (DXIXa).

[0688] [ka]

[0689] In another embodiment, the compound of formula (DXV) has the structure of formula (DXVa):

[0690] [ka]

[0691] In some embodiments, the process for preparing a compound of formula (DII) comprises: structure:

[0692] [ka] providing a compound of formula (DXX) having the formula: forming a compound of formula (DXIX) from a compound of formula (DXX) may include:

[0693] In some embodiments, The step of forming a compound of formula (DXIX) comprises: reacting a compound of formula (DXX) with an LG"-introducing agent to produce a compound of formula (DXIX); Includes:

[0694] In one embodiment, the LG″ introducing agent is NBu4Br3.

[0695] In another embodiment, the compound of formula (DXX) has the structure of formula (DXXa).

[0696] [ka]

[0697] Therapeutic Formulations, Administration, and Uses The present disclosure provides pharmaceutical compositions comprising a peptidomimetic or compound of the present disclosure.

[0698] The compositions of the present disclosure can be formulated with suitable carriers, excipients, and other agents that provide improved migration, delivery, tolerability, etc. Numerous suitable formulations can be found in Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, a formulary known to all pharmaceutical chemists. These formulations include, for example, powders, pastes, ointments, gels, waxes, oils, lipids, lipid (cationic or anionic)-containing vesicles (e.g., LIPOFECTIN™, Life Technologies, Carlsbad, CA), DNA conjugates, anhydrous absorbent pastes, oil-in-water and water-in-oil emulsions, emulsions of carbowax (polyethylene glycols of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax. See also Powell et al., "Compendium of Excipients for Parenteral Formulations," PDA (1998) J Pharm Sci Technol 52:238-311.

[0699] The dose of a peptidomimetic or compound administered to a patient may vary depending on the patient's age and size, the target disease, condition, route of administration, etc. Suitable doses are typically calculated according to body weight or body surface area. When a peptidomimetic or compound of the present disclosure is used for therapeutic purposes in an adult patient, it may be advantageous to administer the peptidomimetic or compound of the present disclosure intravenously, typically at a single dose of about 0.01 to about 20 mg / kg body weight, more preferably about 0.02 to about 7, about 0.03 to about 5, or about 0.05 to about 3 mg / kg body weight. The frequency and duration of treatment may be adjusted depending on the severity of the condition. Effective dosages and schedules for administering a peptidomimetic or compound can be determined empirically; for example, the patient's progress can be monitored by periodic evaluation, and the dosage can be adjusted accordingly. Furthermore, interspecies scaling of dosages can be performed using methods well known in the art (e.g., Mordenti et al., 1991, Pharmaceut. Res. 8:1351).

[0700] Various delivery systems are known and can be used to administer the pharmaceutical compositions of the present disclosure, such as liposomal encapsulation, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, and receptor-mediated endocytosis (see, e.g., Wu et al., 1987, J. Biol. Chem. 262:4429-4432). Methods of introduction include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions can be administered by any convenient route, such as by infusion or bolus injection, or by absorption through epithelial or mucocutaneous linings (e.g., oral, rectal, and intestinal mucosa), and can be administered together with other biologically active agents. Administration can be systemic or local.

[0701] The pharmaceutical compositions of the present disclosure can be delivered subcutaneously or intravenously using a standard needle and syringe. In addition, for subcutaneous delivery, pen delivery devices are easily adapted to deliver the pharmaceutical compositions of the present disclosure. Such pen delivery devices can be reusable or disposable. Reusable pen delivery devices generally utilize a replaceable cartridge containing the pharmaceutical composition. Once all the pharmaceutical composition in the cartridge has been administered and the cartridge is empty, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. This allows the pen delivery device to be reused. Disposable pen delivery devices do not have a replaceable cartridge. Instead, disposable pen delivery devices are provided with a reservoir within the device that holds the pharmaceutical composition pre-filled. Once the reservoir is emptied of the pharmaceutical composition, the entire device is discarded.

[0702] Numerous reusable pens and autoinjector delivery devices are adapted for subcutaneous delivery of the pharmaceutical compositions of the present disclosure. Examples include, but are not limited to, the AUTOPEN™ (Owen Mumford, Inc., Woodstock, UK), the DISETRONIC™ pen (Disetronic Medical Systems, Bergdorf, Switzerland), the HUMALOG MIX 75 / 25™ pen, the HUMALOG™ pen, the HUMALIN 70 / 30™ pen (Eli Lilly and Co., Indianapolis, IN), the NOVOPEN™ I, II, and III (Novo Nordisk, Copenhagen, Denmark), the NOVOPEN JUNIOR™ (Novo Nordisk, Copenhagen, Denmark), the BD™ pen (Becton Dickinson, Franklin Lakes, NJ), the OPTIPEN™, the OPTIPEN PRO™, the OPTIPEN IV ... Examples of disposable pen delivery devices applicable to subcutaneous delivery of the pharmaceutical compositions of the present disclosure include, but are not limited to, the SOLOSTAR™ pen (sanofi-aventis), the FLEXPEN™ (Novo Nordisk), and the KWIKPEN™ (Eli Lilly), the SURECLICK™ autoinjector (Amgen, Thousand Oaks, CA), the PENLET™ (Haselmeier, Stuttgart, Germany), the EPIPEN (Dey, LP), and the HUMIRA™ Pen (Abbott Labs, Abbott Park, IL), to name just a few.

[0703] In certain circumstances, pharmaceutical compositions can be delivered in controlled release systems. In one embodiment, a pump can be used (see Langer, supra; Sefton, 1987, CRC Crit. Ref. Biomed. Eng. 14:201). In another embodiment, a polymeric material can be used; see Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Press, Boca Raton, Florida. In yet another embodiment, a controlled release system can be placed in proximity to the target of the composition, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, 1984, Medical Applications of Controlled Release, supra, Vol. 2, pp. 115-138). Other controlled release systems are discussed in the review by Langer, 1990, Science 249:1527-1533.

[0704] Injectable preparations may include dosage forms for intravenous, subcutaneous, intradermal, and intramuscular injections, infusions, and the like. These injectable preparations may be prepared by known methods. For example, injectable preparations may be prepared by dissolving, suspending, or emulsifying the above-described antibody or its salt in a sterile aqueous or oily medium conventionally used for injections. Aqueous media for injection include, for example, saline, isotonic solutions containing glucose and other auxiliary agents, and the like, which may be used in combination with appropriate solubilizers, such as alcohols (e.g., ethanol), polyalcohols (e.g., propylene glycol, polyethylene glycol), nonionic surfactants (e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)), and the like. Oily media include, for example, sesame oil, soybean oil, and the like, which may be used in combination with solubilizers, such as benzyl benzoate, benzyl alcohol, and the like. The injectable preparations prepared in this manner are preferably filled into appropriate ampoules.

[0705] Advantageously, the above-described pharmaceutical compositions for oral or parenteral use are prepared in a unit dosage form suitable for the dosage of the active ingredient. Such unit dosage forms include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc. The amount of the antibody contained is generally about 5 to about 500 mg per unit dosage form. In particular, it is preferable that the antibody be contained in an amount of about 5 to about 100 mg in the form of an injection, or about 10 to about 250 mg in other dosage forms.

[0706] Therapeutic uses of peptidomimetics or compounds In another embodiment, the peptidomimetics or compounds disclosed herein (e.g., GLP1 peptidomimetics) are useful, inter alia, for the treatment, prevention, and / or amelioration of diseases, disorders, or conditions in need of treatment.

[0707] In one aspect, the present disclosure provides a method for treating a condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a peptidomimetic or compound (e.g., a GLP1 peptidomimetic) according to the present disclosure, or a composition comprising any compound according to the present disclosure.

[0708] In some embodiments, the peptidomimetics or compounds (e.g., GLP1 peptidomimetics) disclosed herein are useful for treating any disease or disorder in which stimulation, activation, and / or targeting of GLP1R is beneficial. In particular, the compounds of the present disclosure can be used for the treatment, prevention, and / or amelioration of any disease or disorder associated with or mediated by GLP1R expression or activity.

[0709] In some embodiments, the peptidomimetics or compounds (e.g., GLP1 peptidomimetics) disclosed herein are useful for treating GLP1R-associated diseases. In some embodiments, the GLP1R-associated disease is type 1 or type 2 diabetes mellitus. The administered peptidomimetic or compound (e.g., GLP1 peptidomimetic) can cause at least one of the following results: induction of insulin secretion, suppression of glucagon release, reduction of blood glucose, improvement of glycemic control, promotion of islet neogenesis and delayed gastric emptying, or enhancement of glucose-resistant islets.

[0710] In some embodiments, the GLP1R-related disease is a neurodegenerative disorder, cognitive impairment, memory impairment, or learning disorder.The neurodegenerative disorder can be, for example, dementia, senile dementia, mild cognitive impairment, Alzheimer's-related dementia, Huntington's chorea, tardive dyskinesia, hyperkinesia, mania, Parkinson's disease, Steele-Richard syndrome, Down syndrome, myasthenia gravis, nerve trauma, brain trauma, vascular amyloidosis, cerebral hemorrhage with amyloidosis I, brain inflammation, Friedreich's ataxia, acute confusional disorder, amyotrophic lateral sclerosis, glaucoma, and Alzheimer's disease.

[0711] In some embodiments, the GLP1R-related disease is a liver disease. The liver disease can be, for example, non-alcoholic fatty liver disease (NAFLD), fatty liver, non-alcoholic steatohepatitis (NASH), and cirrhosis.

[0712] In some embodiments, the GLP1R-related disease is coronary artery disease, which can be, for example, cardiomyopathy and myocardial infarction.

[0713] In some embodiments, the GLP1R-related disease is a kidney disease. The kidney disease can be, for example, hypertension or chronic renal failure.

[0714] In some embodiments, the GLP1R-related condition is an eating disorder. The eating disorder can be, for example, overeating.

[0715] Without being bound by theory, the peptide mimetics or compounds (e.g., GLP1 peptide mimetics) disclosed herein can be used to attenuate the effects of apoptosis-mediated degenerative diseases of the central nervous system, such as Alzheimer's disease, Creutzfeldt-Jakob disease and bovine spongiform encephalopathy, chronic wasting syndrome, and other prion-mediated apoptotic neurological diseases (see, e.g., Perry and Grieg (2004) Current Drug Targets 6:565-571). Administration of the peptide mimetics or compounds (e.g., GLP1 peptide mimetics) disclosed herein can also result in downregulation of βAPP, thereby ameliorating Aβ mono- or oligomer-mediated pathology associated with Alzheimer's disease (see, e.g., Perry et al. (2003) Journal of Neuroscience Research 72:603-612).

[0716] It is also contemplated that the peptide mimetics or compounds (e.g., GLP1 peptide mimetics) disclosed herein can be used to improve learning and memory, for example, by increasing neuronal plasticity and promoting cell differentiation (see During et al. (2003) Nature Medicine 9:1173-1179). Furthermore, the compounds (e.g., antibody-drug conjugates, linker-payloads, and / or payloads) disclosed herein can also be used to preserve dopamine neurons and motor function in Parkinson's disease (see, for example, Greig et al. (2005) Abstract 897.6, Society for Neuroscience, Washington, DC).

[0717] In some embodiments, the peptidomimetics or compounds (e.g., GLP1 peptidomimetics) disclosed herein can also be used to treat metabolic disorders, such as obesity, dyslipidemia, metabolic syndrome X, and pathologies manifested by islet dysfunction.

[0718] Additional diseases that can be treated with the compounds (e.g., antibody-drug conjugates, linker-payloads, and / or payloads) of the present disclosure include autoimmune diseases, particularly those associated with inflammation, including, but not limited to, autoimmune diabetes, adult-onset diabetes, morbid obesity, metabolic syndrome X, and dyslipidemia. For example, anti-GLP1R antibody-drug conjugates can be used as growth factors to promote islet growth in humans with autoimmune diabetes. The compounds (e.g., antibody-drug conjugates, linker-payloads, and / or payloads) described herein can also be useful for treating congestive heart failure.

[0719] In one aspect, the present disclosure provides a method for selectively targeting an antigen (e.g., GLP1R) on the surface of a cell using a peptidomimetic or compound disclosed herein. In one embodiment, the method for selectively targeting an antigen (e.g., GLP1R) on the surface of a cell using a compound comprises linking the compound to a targeted antibody. In one embodiment, the cell is a mammalian cell. In one embodiment, the cell is a human cell. In one embodiment, the cell is a pancreatic cell or a brain cell. [Example]

[0720] The following examples illustrate certain aspects of the present invention, and should not be construed as limiting, as the examples are provided solely for the purpose of providing a more specific understanding and implementation of the embodiments and their various aspects.

[0721] Abbreviations used in the examples and throughout the specification are as follows:

[0722] [Table 1-1]

[0723] [Table 1-2]

[0724] Table 1-3

[0725] Table 1-4

[0726] Table 1-5

Example

[0727] General methods All anhydrous reactions were carried out in oven-dried glassware under an argon atmosphere with mechanical or magnetic stirring unless otherwise noted. Rink Amide Linker was supplied by GL Biochem. All other reagents and anhydrous solvents were purchased from commercial sources and used without further purification unless otherwise noted. Reactions (when possible) were monitored using thin-layer chromatography (TLC) analysis on silica gel 60 F254 precoated plates, visualized by short-wave ultraviolet irradiation (254 nm, where applicable), stained with PMA (5% wt / v) in EtOH or ninhydrin (5% wt / v) in EtOH, followed by carbonization at approximately 200 °C. Column chromatography (silica gel 60, 40 mm) was performed in air and under pressure (0.1–0.3 bar) unless otherwise specified. Bulk solvents for flash chromatography (EtOAc, hexane, THF, DCM) were distilled before use. IR spectra were recorded on an FT-IR spectrometer (Thermo NICOLET iS10) using diamond head sampling and are reported in wavenumbers (cm). Residual solvent content was recorded by GC (SHIMADZU-GC2030 & HS-20). Water content was recorded on a Mettler Toledo C30s Karl Fischer reactor. Melting points (MP) were measured by DSC (differential scanning calorimetry, DSC214). Inorganic salt content was provided by a Thermo ICS-6000 ion chromatography detector. Purity of synthetic intermediates after chromatographic purification was determined to be >90% by 1H NMR spectroscopy. Purity of the final compounds was ≥95% (NMR analysis) after trituration with MeOH and MeCN (Honeywell, HPLC grade).

[0728] General NMR Methods and Instrumentation 1 H NMR and 13C: 1 H NMR spectra were obtained using a Bruker Avance 400 (400 MHz, 1 H, 100MHz, 13C) recorded on a spectrometer. Chemical shift values ​​(δ) are reported in ppm relative to residual chloroform ( 1 H: δ 7.26 ppm; 13 C, δ 78.0 ppm). Multiplicities are designated as s (singlet), d (doublet), t (triplet), q (quartet), p (quintet), h (sextet), dd-doublet doublet, dt-triplet doublet, dq-quartet doublet, m (multiplet), and br (broadband). 1 H and 13 To confirm the C signal, 1 H, 13 Coupling constants (J) are reported in Hertz (Hz). [Example]

[0729] General LC-MS methods and equipment Method 1 This method was used to analyze intermediates 1, 2, 3, 4, 5, 6, and 7 of tag 5. Instrument: Agilent 1200+6110MS (Table 1). An Xbridge Protein BEH C4 300A 2.5µm 4.6 x 50mm column was used for chromatography. Diode array (DAD) and evaporative light scattering (ELSD) were used as detection methods. The MS mode was positive electrospray ionization. The MS range was 50-1500. Mobile phase A was 0.04% TFA in water, and mobile phase B was 0.02% TFA in HPLC-grade ACN. The gradient was 50–100% B for 6 min, 50% B at 0.00 min, 50–100% B from 0.00–4.00 min, hold at 100% B for 1.5 min, 100–50% B from 5.51–6.00 min, hold at 50% B for 0.49 min. The flow rate was 1.50 mL / min.

[0730] [Table 2]

[0731] Method 2 This method was used for the analysis of intermediate 8 of tag 5. Instrument: Waters H Class & SQD2 (Table 2). The gradient was 50-100% B at 3.40 min, hold at 100% B at 3.40-3.85 min, hold at 100% B at 0.01 min, hold at 5% at 0.64 min. The flow rate was 0.80 ml / min. Mobile phase A was H2O + 10 mM NH4HCO3, and mobile phase B was HPLC grade THF. The column used for chromatography was 2.1 * The chromatogram was analyzed on a 50 mm Xbridge Shield RPC18 column (5 μm particles). Diode array (DAD) detection and positive electrospray ionization were used as the detection methods. The MS range was 100–3000 Da.

[0732] [Table 3]

[0733] Method 3 This method was used for the analysis of intermediate 9 and tag 5. Instrument: Waters H Class & SQD2 (Table 3). The gradient was 70-100% B at 16:00 min, hold at 100% B at 4:00 min, 100-70% B at 0.01 min, hold at 70% for 3 min, and the flow rate was 1.0 ml / min. Mobile phase A was 10 mM ammonium bicarbonate, and mobile phase B was HPLC-grade THF. The column used for chromatography was an Xbridge C18 4.6 * The column was 150 mm (3.5 μm particles). Diode array (DAD) and positive electrospray ionization were used as detection methods. The MS range was 100–3000 Da.

[0734] [Table 4]

[0735] Method 4 This method was used to analyze Tag 1-TCP (Tag 5-1AA). LC-MS conditions: Mobile phase: 1.5 mL / 4 L TFA in water (solvent A) and 0.75 mL / 4 L TFA in ACN (solvent B), with a flow rate of 0.8 mL / min and an elution gradient of 50% to 100% (solvent B) for 1.35 min, held at 100% for 0.9 min; Column: Xtimate C18 2.1 * 30mm, 3um.

[0736] Method 5 This method was used to analyze Tag 1-2AA, Tag 1-3AA, Tag 1-4AA, Tag 1-5AA, Tag 1-6AA, Tag 1-7AA, and Tag 1-10AA. LC-MS conditions: Mobile phase: 1.5 mL / 4 L TFA in water (solvent A) and 0.75 mL / 4 L TFA in ACN (solvent B), with a flow rate of 0.8 mL / min and a gradient from 10% to 80% (solvent B) for 6 min, with a hold at 80% for 0.5 min. Column: Xtimate C18 2.1 * 30mm, 3um.

[0737] Method 6 This method was used for the analysis of M2. LC-MS conditions: Reverse-phase LC-MS was performed using a Chromolith Flash RP-C18 25-3 mm column eluted with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min.

[0738] Method 7 This method was used to analyze Tag 5-2AA, Tag 5-3AA, Tag 5-4AA, Tag 5-5AA, Tag 5-6AA, Tag 5-7AA, Tag 5-8AA, Tag 5-9AA, and Tag 5-10AA. LCMS conditions (BF): Mobile phase: 1.5 mL / 4 L TFA in water (solvent A) and 0.75 mL / 4 L TFA in ACN (solvent B), with a flow rate of 0.8 mL / min and an elution gradient of 10% to 80% (solvent B) for 6 min, with a hold at 80% for 0.5 min; Column: Xtimate C18 2.1 * 30mm, 3um; Wavelength: UV220nm, 254nm; Column temperature 50℃; MS ionization: ESI.

[0739] Method 8 This method was used to analyze Tag 5-2AA, Tag 5-3AA, Tag 5-4AA, Tag 5-5AA, Tag 5-6AA, Tag 5-7AA, Tag 5-8AA, Tag 5-9AA, and Tag 5-10AA. LCMS conditions (CR): Mobile phase: 1.5 mL / 4 L TFA in water (solvent A) and 0.75 mL / 4 L TFA in ACN (solvent B), flow rate: 0.8 mL / min, elution gradient: 10% to 80% (solvent B) for 6 min, hold at 80% for 0.5 min; Column: NanoChrom ChromCore 120 C18 3µm, 3.0 * 30mm; Wavelength: UV220nm, 254nm; Column temperature 50℃; MS ionization: ESI.

[0740] Method 9 This method was used to analyze Tag 5-2AA, Tag 5-3AA, Tag 5-4AA, Tag 5-5AA, Tag 5-6AA, Tag 5-7AA, Tag 5-8AA, Tag 5-9AA, and Tag 5-10AA. LCMS conditions (BV): Mobile phase: 1.5 mL / 4 L TFA in water (solvent A) and 0.75 mL / 4 L TFA in ACN (solvent B), with a flow rate of 0.8 mL / min and an elution gradient of 10% to 80% (solvent B) for 6 min, with a hold at 80% for 0.5 min; Column: NanoChrom ChromCore 120 C18 3µm, 3.0 * 30 mm; wavelength: UV 220 nm; column temperature 50°C; MS ionization: ESI.

[0741] Method 10 This method was used for the analysis of Tag 5-2AA, Tag 5-3AA, Tag 5-4AA, Tag 5-5AA, Tag 5-6AA, Tag 5-7AA, Tag 5-8AA, Tag 5-9AA, and Tag 5-10AA. LCMS conditions (BK): Instrument and column: Xtimate 3um, C18, 2.1 *30mm S / N3U411701820 Mobile phase: 1.5mL / 4L TFA in water (solvent A) and 0.75mL / 4L TFA in ACN (solvent B) with a flow rate of 0.8ml / min and a gradient of 10% to 80% (solvent B) for 6 minutes, held at 80% for 0.5 minutes. ESI source: positive ion mode, wavelengths 220nm and 254nm, oven temperature 50°C.

[0742] Method 11 This method was used for the analysis of tag 4-10AA. HPLC Method A: Column: YMC-Pack ODS-A 150 * 4.6 mm, 5 μm; 2.75 mL / 4 L TFA in water (solvent A) and 2.5 mL / 4 L TFA in ACN (solvent B) at a flow rate of 1.5 mL / min. Use an elution gradient of 10% to 80% (solvent B) for 10 min, hold at 80% for 5 min.

[0743] Method 12 This method was used for the analysis of Tag 4-10AA. LCMS conditions: Reverse-phase LCMS was performed using a Chromolith Flash RP-C18 25-3 mm column eluted with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min.

[0744] Method 13 This method was used to analyze the intermediates AA10-AA9-AA8-OH. LC-MS conditions: Mobile phase: 1.5 mL / 4 L TFA in water (solvent A) and 0.75 mL / 4 L TFA in ACN (solvent B), flow rate 1.5 mL / min, elution gradient 5% to 95% (solvent B) in 0.7 min, hold 95% for 0.4 min; Column: Agilent Pursult 5 C18 20 * 2.0mm.

[0745] Method 14 This method was used for the analysis of DMB-AA9. Reverse phase HPLC was performed using Ultimate XB-C18, 3 μm, 3.0 *A 50 mm column was used. Mobile phase: 1.0% ACN (0.1% TFA) in water to 5% ACN (0.1% TFA) in water over 1 min, then 5% ACN (0.1% TFA) in water to 100% ACN (0.1% TFA) over 5 min, hold at 100% ACN (0.1% TFA) for 2 min, return to 1.0% ACN (0.1% TFA) in water over 8.01 min, hold for 2 min. Flow rate: 1.2 ml / min.

[0746] Method 15 This method was used for the analysis of M2. HPLC conditions: Mobile phase: 1.0% ACN (0.1% TFA) in water to 5% ACN (0.1% TFA) in water in 1 minute, then 5% ACN (0.1% TFA) in water to 100% ACN (0.1% TFA) in 5 minutes, hold at 100% ACN (0.1% TFA) for 2 minutes, then return to 1.0% ACN (0.1% TFA) in water in 8.01 minutes and hold for 2 minutes. Flow rate: 1.2 ml / min. Column: Ultimate XB-C18, 3 um, 3.0 * 50mm.

[0747] ELSD LCMS conditions Mobile phase: 10 mM NHOAc in water (solvent A) and ACN (solvent B). Flow rate: 0.8 mL / min. Elution gradient: 10% to 80% (solvent B) for 5 min, hold at 80% for 1.95 min, change 80% B to 10% B for 7 min, hold at 10% for 4 min. Column: Shim-pack Scepter C18-120 1.9 μm, 2.1 * 100mm. [Example]

[0748] Synthesis of DMB-AA9 Scheme 1 below depicts the synthesis of DMB-AA9.

[0749] Scheme 1

[0750] [ka] Conditions: 1) (2,4-Dimethoxyphenyl)methanamine (1.2 eq.), HATU (1.0 eq.), DIPEA (3 eq.), DMF (100 mL), 20 °C, 2 h, 84% yield; 2) (a) PCl (1.5 eq.), TMSN (4 eq.), pyridine (8V), 0 °C, 4 h; (b) MeOH (4 eq.), 0 °C, 2 h, 81% yield; 3) (a) NaOH (3 eq.), THF / HO (2:1, 10 mL), 20 °C, 2 h; (b) FmocOSu (1 eq.), aq. NaHCO, 20 °C, 2 h; 40.5% yield.

[0751] Step 1: Synthesis of methyl (2S)-4-[(2,4-dimethoxyphenyl)methylamino]-2-(9H-fluoren-9-ylmethoxycarbonylamino)-4-oxo-butanoate (DMB-AA9-2)

[0752] [ka] To a solution of (3S)-3-(9H-fluoren-9-ylmethoxycarbonylamino)-4-methoxy-4-oxobutanoic acid (DMB-AA9-1, 90 g, 243.66 mmol, 1 eq.) in DMF (200 mL), HATU (101.91 g, 268.03 mmol, 1.1 eq.) and DIPEA (62.98 g, 487.32 mmol, 84.88 mL, 2 eq.) were added, and the mixture was stirred at 20 °C for 30 min. Then, (2,4-dimethoxyphenyl)methanamine (44.82 g, 268.03 mmol, 40.37 mL, 1.1 eq.) was added, and the mixture was stirred at 20 °C for 1 hr. Most of the reactant was consumed by TLC (PE / EtOAc = 2 / 1), and the desired compound was detected by LCMS. The reaction mixture was poured into DCM (1500 mL) and washed with H2O (900 mL) and saturated citric acid (100 mL). The separated organic layer was washed with brine (1000 mL), dried over Na2SO4, and filtered. It was then concentrated under reduced pressure to give the crude product as a light yellow solid. The crude product was triturated with EtOAc (600 mL) for 12 hours and filtered to give the target compound. The desired compound DMB-AA9-2 (109 g, 197.59 mmol, 81.09% yield, 94% purity) was obtained as a white solid. ESI-MS (Method 10): RT = 3.896 min, C 29 H 30 Calculated mass of O7N2H: 519.2, measured m / z: 519.1 [M+H] + . 1H NMR (400MHz, chloroform-d) δ = 7.76 (d, J=7.5 Hz, 2H), 7.61 (br dd, J=3.6, 6.9 Hz, 2H), 7.44 - 7.37 (m, 2H), 7.36 - 7.28 (m, 2H), 7.15 (d, J=8.3 Hz, 1H), 6.48 - 6.38 (m, 2H), 6.15 (br s, 2H), 4.60 (br s, 1H), 4.44 - 4.26 (m, 4H), 4.26 - 4.19 (m, 1H), 3.84 - 3.72 (m, 9H), 2.97 (br dd, J=4.4, 15.7 Hz, 1H), 2.73 (br dd, J=3.9, 15.7 Hz, 1H).

[0753] Step 2: Synthesis of methyl (2S)-3-[1-[(2,4-dimethoxyphenyl)methyl]tetrazol-5-yl]-2-(9H-fluoren-9-ylmethoxycarbonylamino)propanoate (DMB-AA9-3)

[0754] [ka] To a suspension of compound DMB-AA9-2 (100 g, 192.84 mmol, 1 eq.) in pyridine (800 mL), TMSN3 (88.87 g, 771.37 mmol, 101.45 mL, 4 eq.) was added, followed by portionwise addition of PCl5 (60.24 g, 289.26 mmol, 1.5 eq.). The temperature was maintained below 0 °C using an ice-brine bath. The reaction mixture was stirred at 0 °C for 5 h. LCMS showed that most of the reactant was consumed. Then, MeOH (24.71 g, 771.37 mmol, 31.21 mL, 4 eq.) was added dropwise, and the reaction mixture was stirred at 0 °C for 1 h. TLC (PE / EtOAc = 2 / 1) and LCMS showed that most of the reactant was consumed, and the desired compound was detected as the main peak. The reaction was quenched with MeOH (20 mL) and saturated aqueous NaHCO3 (1000 mL x 2) and extracted with EtOAc (1000 mL x 3). The aqueous layer was quenched with 8-10% NaClO (1000 mL). The combined organic layers were concentrated under reduced pressure to give a residue, which was redissolved in EtOAc (2000 mL), washed with saturated citric acid (500 mL x 2) and brine (1000 ml), dried over Na2SO4, and concentrated under reduced pressure to give a residue. The crude product was used directly in the next step without further purification. The desired compound DMB-AA9-3 (97 g, 156.86 mmol, 81.34% yield, 87.9% purity) was obtained as a yellow oil. LC-MS: RT = 4.310 min, C 29 H 29 Calculated mass of N5O6H: 544.2 [M+H] + , m / z Measured value 544.2 [M+H] + , Method 10. 1H NMR (400MHz, chloroform-d) δ = 7.68 (d, J=7.6 Hz, 2H), 7.48 (br d, J=7.4 Hz, 2H), 7.37 - 7.25 (m, 3H), 7.25 - 7.20 (m, 2H), 7.09 (d, J=8.3 Hz, 1H), 6.43 - 6.31 (m, 2H), 6.09 (br d, J=8.5 Hz, 1H), 4.90 - 4.75 (m, 1H), 4.36 - 4.22 (m, 2H), 4.20 - 4.10 (m, 1H), 3.71 - 3.63 (m, 9H), 3.54 - 3.41 (m, 1H), 3.26 (dd, J=4.6, 16.3 Hz, 1H) ppm.

[0755] Step 3: Synthesis of (2S)-3-[1-[(2,4-dimethoxyphenyl)methyl]tetrazol-5-yl]-2-(9H-fluoren-9-ylmethoxycarbonylamino)propanoic acid (DMB-AA9)

[0756] [ka] To a solution of compound DMB-AA9-3 (95 g, 174.77 mmol, 1 eq) in THF (600 mL) and HO (100 mL), NaOH (20.97 g, 524.31 mmol, 3 eq) was added, and the reaction mixture was stirred at 20 °C for 2 h. LCMS showed that the reactant was consumed. The pH of the reaction mixture was adjusted to pH 7–8 using citric acid, and the mixture was diluted with HO (200 mL) and then extracted with TBME (300 mL × 2). To the aqueous layer, NaHCO (44.05 g, 524.31 mmol, 20.39 mL, 3 eq) and FmocOSu (58.96 g, 174.77 mmol, 1 eq) in THF (600 mL) were added, and the reaction mixture was stirred at 10 °C for 12 h. LCMS showed that the desired compound was detected as the main peak. The reaction was concentrated under reduced pressure to remove THF, then poured into saturated NaHCO3 (500 mL) and H2O (300 mL) and extracted with TBME (500 mL x 3). The aqueous layer was extracted with EtOAc / MeOH (10 / 1, 800 mL x 3). The combined EtOAc layers were washed with saturated NaHCO3 (200 mL) and brine (200 ml), then dried over Na2SO4 and concentrated under reduced pressure to give a residue. The crude product was triturated with TBME:ACN:EtOAc (10:1:1, 250 mL) at 20 °C for 12 h and filtered to give a light yellow solid. Compound DMB-AA9 (38 g, 70.90 mmol, 40.57% yield, 98.8% purity) was obtained as a light yellow solid. LC-MS: RT = 3.207 min, C 28 H 27 Calculated mass of N5O6H: 530.2 [M+H] + , m / z measured value 530.2 [M+H] + , Method 7. HPLC purity: 98.84% after grinding (Method 14 was used for the analysis of DMB-AA9). 1H NMR (400MHz, methanol-d4) δ = 7.77 (d, J=7.5 Hz, 2H), 7.62 (br d, J=7.3 Hz, 2H), 7.40 - 7.16 (m, 5H), 6.56 - 6.43 (m, 2H), 5.52 - 5.37 (m, SFC ee = 98.53-1.47 = 97.06%. [Example]

[0757] Synthesis of M2 onto Tag1 (LPPS) 4.1 Tag1 Synthesis The synthetic route to make Tag 1 is shown in Scheme 2.

[0758] Scheme 2

[0759] [ka] Reagents and conditions: a) 1-bromodocosane (2.2 eq.), K2CO3 (6 eq.), DMF / THF, 90 °C, 12 h, 99%; b) NaBH4 (5 eq.), EtOH (1 V), THF (10 V), 70 °C, 12 h, 96%; c) FmocNH2 (2 eq.), MeSO3H (0.3 eq.), toluene / THF, 110 °C, 36 h, 82%; d) 20% piperidine / THF (10 V), 20 °C, 2 h, 92%. Total yield: 71.6% over four steps.

[0760] This synthesis was reported by Hirai et al. in U.S. Patent Application Publication No. 2012 / 0296074. Alkylation of tag 1-1 with 1-bromodocosane gave tag 1-2, which was reduced with sodium borohydride to form alcohol tag 1-3. Compound tag 1-4 was obtained from tag 1-3 by nucleophilic substitution of 9-fluorenylmethyl carbamate using methanesulfonic acid as a catalyst. The reaction yield was slightly improved (82% vs. 60%) by using a cosolvent (toluene / THF) instead of toluene. Fmoc deprotection of tag 1-4 was carried out using piperidine / THF to give tag 1 in a total yield of 72%. Hirai et al. also reported a 45% yield by Fmoc deprotection of tag 1-4 using DBU / DCM.

[0761] Step a: Synthesis of bis(4-docosoxyphenyl)methanone (TAG1-2)

[0762] [ka] To a mixture of compound Tag 1-1 (10 g, 46.68 mmol, 1.0 eq.) and 1-bromodocosane (40.00 g, 102.70 mmol, 2.2 eq.) in DMF (200 mL) and THF (40 mL), K2CO3 (38.71 g, 280.09 mmol, 6 eq.) was added, and the mixture was stirred at 90 °C for 12 h. The reactant was consumed, and a new, less polar spot formed on TLC (PE / EtOAc = 2 / 1). The reaction mixture was poured into water (1000 mL), and the mixture was stirred for 1 h. The precipitated solid was then collected by filtration and slurry washed with water (200 mL × 2). The resulting solid was triturated with acetone (500 mL) and collected by filtration of the acetone-washed slurry. After filtration, the resulting solid was dried under reduced pressure to give tag 1-2 (38.8 g, 46.67 mmol, 99.97% yield) as a white solid.

[0763] Compound tag 1-2 (C 57 H 98 O3, MW:830.75):1 H NMR (400MHz, CDCl3) δ = 7.78 (d, J=8.8 Hz, 4H), 6.95 (d, J=8.8 Hz, 4H), 4.04 (t, J=6.5 Hz, 4H), 1.86 - 1.78 (m, 4H), 1.51 - 1.43 (m, 4H), 1.26 (s, 72H), 0.89 (t, J=6.8 Hz, 6H) ppm.

[0764] Step b: Synthesis of bis(4-docoxyphenyl)methanol (TAG 1-3)

[0765] [ka] A solution of compound Tag 1-2 (35 g, 42.10 mmol, 1 eq.) in THF (500 mL) and EtOH (25 mL) was heated to 70 °C. Then, NaBH (7.96 g, 210.49 mmol, 5.0 eq.) was slowly added, and the mixture was stirred at the same temperature for 20 h. TLC (PE / EtOAc = 3 / 1) showed that the reactant was completely consumed, and a new spot was detected. The reaction mixture was cooled on ice, and 1 N hydrochloric acid (80 mL) was added dropwise. THF was evaporated, water (450 mL) was added, and the pH was adjusted to 5-7 with 1 N hydrochloric acid. The slurry was filtered, and the resulting crystals were washed with water and methanol to give product Tag 1-3 (34 g, 40.80 mmol, 96.91% yield) as a white solid.

[0766] Compound tags 1-3 (C 57 H 100 O3, MW:832.77): 1H NMR (400MHz, chloroform-d) δ = 7.29 - 7.25 (m, 4H), 6.88 - 6.84 (m, 4H), 5.77 (d, J=2.8 Hz, 1H), 3.94 (t, J=6.7 Hz, 4H), 2.09 (d, J=3.3 Hz, 1H), 1.81 - 1.73 (m, 4H), 1.48 - 1.41 (m, 4H), 1.35 - 1.26 (m, 72H), 0.91 - 0.87 (m, 6H) ppm.

[0767] Step c: Synthesis of 9H-fluoren-9-ylmethyl N-[bis(4-docosoxyphenyl)methyl]carbamate (Tags 1-4)

[0768] [ka] To a mixture of tag 1-3 (10 g, 12.00 mmol, 1 eq.) and 9H-fluoren-9-ylmethylcarbamate (FmocNH, 5.74 g, 24.00 mmol, 2.0 eq.) in toluene (200 mL), MsOH (345.97 mg, 3.60 mmol, 256.27 μL, 0.3 eq.) was added, and the mixture was stirred at 110 °C for 36 h. Most of the reactant was consumed, and a new spot formed on TLC (PE / EtOAc = 10 / 1). The reaction mixture was poured into 5% NaHCO (30 mL), and the mixture was stirred for 1 h. The precipitated solid was then collected by filtration and washed with water (50 mL). The resulting solid was triturated with ACN (200 mL), and the solid was collected by filtration and slurry washed with methanol. After filtration, the resulting solid was dried under reduced pressure to give tag 1-4 (11 g, 9.91 mmol, 82.58% yield, 95% purity) as an off-white solid.

[0769] Compound tags 1-4 (C 72 H 111 NO4, MW: 1053.85): 1H NMR (400MHz, chloroform-d) δ = 9.89 (s, 1H), 7.78 (br d, J=8.4 Hz, 2H), 7.61 (br d, J=7.4 Hz, 2H), 7.44 - 7.31 (m, 4H), 7.14 - 7.10 (m, 1H), 7.12 (d, J=8.6 Hz, 2H), 6.98 (dd, J=8.7, 19.2 Hz, 1H), 6.88 - 6.75 (m, 4H), 5.87 (br s, 1H), 4.43 (d, J=7.0 Hz, 1H), 4.28 - 4.21 (m, 1H), 4.04 (dt, J=2.9, 6.5 Hz, 1H), 3.99 - 3.85 (m, 4H), 1.82 - 1.74 (m, 4H), 1.59 (br s, 4H), 1.27 (s, 72H), 0.91 - 0.89 (m, 6H) ppm.

[0770] Step d: Synthesis of bis(4-docoxyphenyl)methanamine (TAGs 1-5)

[0771] [ka] To a solution of Tag 1-4 (11 g, 10.43 mmol, 1 eq.) in THF (80 mL) was added piperidine (20 mL). The mixture was then stirred at 20 °C for 2 h. TLC (PE / EtOAc = 3 / 1) showed that the reactant was completely consumed and one new spot was detected. ACN (400 mL) was added to the reaction solution, and the resulting suspension was filtered and washed with ACN (100 mL × 2) to give an off-white solid. The crude product Tag 1-5 (8 g, 9.61 mmol, 92.14% yield) was obtained as a white solid. It was dried under reduced pressure and used in the synthesis of Tag 1-M2 below.

[0772] Compound tags 1-5 (C 57 H 101 NO2, MW:831.78): 1H NMR (400MHz, chloroform-d) δ = 7.29 (s, 4H), 6.86 (d, J=8.6 Hz, 4H), 5.15 (s, 1H), 3.95 (t, J=6.6 Hz, 4H), 1.80 - 1.76 (m, 4H), 1.49 - 1.43 (m, 4H), 1.29 (s, 72H), 0.91 (t, J=6.8 Hz, 6H) ppm.

[0773] 4.2 Synthesis of Tag1-M2 Figure 2 and Table 4 outline the synthesis of M2 using Tag1.

[0774] [Table 5]

[0775] General Procedure A for the Amide Coupling Reaction to Generate N-Fmoc-Tagged 1-nAA To a solution of Tag1 or Tag1-(n-1)AA (1 eq.) and AAn (1.5 eq.) in DCM (20 V) was added DIC (1.5 eq.) and HOAt (1.5 eq.). The solution was stirred at 20 °C for 2 h. TLC (PE / EtOAc = 1 / 1) showed complete consumption of the reactant, and a new spot was detected, which was further confirmed by LC-MS after a small amount of cleavage. ACN (80 V) was added to the reaction solution, and the resulting suspension was filtered and washed with MeOH (20 V × 2) to give an off-white solid. The crude product was dried under reduced pressure to give N-Fmoc-Tag1-(n)AA as a white solid, which was used directly in the next step.

[0776] General Procedure B for Fmoc Deprotection to Obtain Tag 1-nAA A solution of N-Fmoc-tag 1-(n)AA (1 eq.) in 20% piperidine in THF (20 V) was stirred at 20 °C for 1 h. TLC (PE / EtOAc = 1 / 1) showed that the reactant was completely consumed and the desired spot was detected. ACN (80 V) was added to the reaction solution, and the resulting suspension was filtered and washed with ACN (20 V × 2) to give an off-white solid. The crude product, Tag 1-nAA, was dried under reduced pressure and used directly in the next step.

[0777] Step 1: Synthesis of (2S)-2-amino-N-[bis(4-docoxyphenyl)methyl]-5-(3,5-dimethylphenyl)pentanamide (TAG1-1AA)

[0778] [ka] To a solution of Tag1 (6 g, 7.21 mmol, 1 eq.) and AA1 (4.80 g, 10.81 mmol, 1.5 eq.) in DCM (100 mL) was added DIC (1.18 g, 9.37 mmol, 1.45 mL, 1.3 eq.) and HOAt (1.28 g, 9.37 mmol, 1.31 mL, 1.3 eq.). The solution was stirred at 20 °C for 2 h. TLC (PE / EtOAc = 1 / 1) showed complete consumption of the reactant and a new spot was detected, which was further confirmed by testing the cleaved crude material. ACN (400 mL) was added to the reaction solution, and the resulting suspension was filtered and washed with MeOH (100 mL × 2) to give an off-white solid. The crude product was dried under reduced pressure to give N-Fmoc-tagged 1-1AA (8 g, 6.36 mmol, 88.23% yield) as a white solid, which was used directly in the next step. LCMS (ESI): RT = 4.266 min, C 28 H 31 N2O3 + Calculated mass 443.23, observed m / z 443.20 [M-tag-1+H] + ;(Tag1=C 57 H 99 O2, MS=815.76).

[0779] To a solution of N-Fmoc-tag 1-1AA (8 g, 6.36 mmol, 1 eq.) in THF (80 mL) was added piperidine (20 mL). The mixture was then stirred at 20 °C for 1 hr. TLC (PE / EtOAc = 1 / 1) showed that the reactant was completely consumed and the desired spot was detected. ACN (400 mL) was added to the reaction solution, and the resulting suspension was filtered and washed with ACN (100 mL × 2) to give an off-white solid. The crude product, tag 1-1AA, was dried under reduced pressure and used directly in the next step.

[0780] Step 2: Synthesis of (2S)-2-[[(2S)-2-amino-3-[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]propanoyl]amino]-N-[bis(4-docosoxyphenyl)methyl]-5-(3,5-dimethylphenyl)pentanamide (TAG1-2AA)

[0781] [ka] The reaction was carried out according to general procedure A for amide coupling using Tag 1-1AA (7 g, 6.76 mmol). The crude product N-Fmoc Tag 1-2AA (11 g, white solid) was dried and used directly in the next step. LCMS (ESI): RT = 5.759 min, C 49 H 55 N6O5 + Calculated mass 807.42, observed m / z 807.40 [M-tag-1+H] + ;(Tag1=C 57 H 99 O2, MS=815.76).

[0782] The deprotection reaction was carried out according to general procedure B for Fmoc deprotection. Tag 1-2AA (8 g, 5.71 mmol, 84.27% yield) was obtained as a white solid. LCMS (ESI): RT = 3.568 min, C 34 H 45 N6O3 + Calculated mass 585.35, observed m / z 585.50 [M-tag-1+H] +;(tag-1=C 57 H 99 O2, MS=815.76).

[0783] Step 3: Synthesis of tert-butyl (3S)-3-amino-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[bis(4-docosoxyphenyl)methylcarbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG1-3AA)

[0784] [ka] The reaction was carried out according to general procedure A for amide coupling using Tag 1-2AA (7 g, 5.00 mmol). N-Fmoc-Tag 1-3AA (8.9 g, 4.96 mmol, 99.25% yield) was obtained as a yellow solid. LCMS (ESI): RT = 5.388 min, C 53 H 60 N7O8 + Calculated mass 922.44, observed m / z 922.50 [M-tag-1+H] + ;(Tag1=C 57 H 99 O2, MS=815.76).

[0785] The deprotection reaction was carried out according to general procedure B for Fmoc deprotection. Tag 1-3AA (6.5 g, 4.14 mmol, 83.36% yield) was obtained as a white solid. LCMS (ESI): RT = 4.294 min, C 38 H 50 N7O6 + Calculated mass 700.37, observed m / z 700.50 [M-tag-1+H] + ;(tag-1=C 57 H 99 O2, MS=815.76).

[0786] Step 4: Synthesis of tert-butyl (3S)-3-[[(2S)-2-amino-3-tert-butoxy-propanoyl]amino]-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[bis(4-docosoxyphenyl)methylcarbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG 1-4AA)

[0787] [ka] The reaction was carried out according to general procedure A for amide coupling using Tag 1-3AA (6.5 g, 4.14 mmol, 1 eq.). N-Fmoc-Tag 1-4AA (8 g, 4.13 mmol, 99.86% yield) was obtained as a yellow solid. LCMS (ESI): RT = 5.198 min, C 56 H 65 N8O 10 + Calculated mass 1009.47, observed m / z 1009.60 [M-tag-1+H] + ;(Tag1=C 57 H 99 O2, MS=815.76).

[0788] The deprotection reaction was carried out according to general procedure B for Fmoc deprotection. Tag 1-4AA (6.6 g, 3.85 mmol, 93.19% yield) was obtained as a white solid. LCMS (ESI): RT = 3.540 min, C 41 H 55 N8O8 + Calculated mass 787.41, observed m / z 787.50 [M-tag-1+H] + ;(Tag1=C 57 H 99 O2, MS=815.76).

[0789] Step 5: Synthesis of tert-butyl (3S)-3-[[(2S)-2-[[(2S,3R)-2-amino-3-tert-butoxy-butanoyl]amino]-3-tert-butoxy-propanoyl]amino]-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[bis(4-docosoxyphenyl)methylcarbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG 1-5AA)

[0790] [ka] The reaction was carried out according to general procedure A for amide coupling using Tag 1-4AA (6.60 g, 3.85 mmol, 1 eq.). N-Fmoc-Tag 1-5AA (8.0 g, 3.82 mmol, 99.25% yield) was obtained as a white solid. LCMS (ESI): RT = 5.024 min, C 60 H 72 N9O 12 + Calculated mass 1110.52, observed m / z 1110.60 [M-tag-1+H] + ;(Tag1=C 57 H 99 O2, MS=815.76).

[0791] The deprotection reaction was carried out according to general procedure B for Fmoc deprotection. Tag 1-5AA (7.0 g, 3.74 mmol, 97.89% yield) was obtained as a white solid, which was used directly in the next step. LCMS (ESI): RT = 3.542 min, C 45 H 62 N9O 10 + Calculated mass 888.45, observed m / z 888.50 [M-tag-1+H] + ;(Tag1=C 57 H 99 O2, MS=815.76).

[0792] Step 6: Synthesis of tert-butyl (3S)-3-[[(2S)-2-[[(2S,3R)-2-[[2-amino-3-(2-fluorophenyl)-2-methyl-propanoyl]amino]-3-tert-butoxy-butanoyl]amino]-3-tert-butoxy-propanoyl]amino]-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[bis(4-docosoxyphenyl)methylcarbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG 1-6AA)

[0793] [ka] The reaction was carried out according to general procedure A for amide coupling using tag 1-5AA (7.00 g, 3.74 mmol, 1 eq.). N-Fmoc tag 1-6AA (7.3 g, 3.21 mmol, 85.87% yield) was obtained as a white solid. LCMS (ESI): RT = 5.024 min, C 70 H 82 FN 10 O 13 + Calculated mass 1289.60, observed m / z 1289.60 [M-tag-1+H] + ;(Tag1=C 57 H 99 O2, MS=815.76).

[0794] The deprotection reaction was carried out according to general procedure B for Fmoc deprotection. Tag 1-6AA (6.5 g, 3.17 mmol, 98.69% yield) was obtained as a white solid. LCMS (ESI): RT = 3.699 min, C 55 H 72 FN 10 O 11 + Calculated mass 1067.53, observed m / z 1067.60 [M-tag-1-3 t Bu+4H] + ;(tag-1=C 57 H 99O2, MS=815.76).

[0795] Step 7: Synthesis of tert-butyl (3S)-3-[[(2S)-2-[[(2S,3R)-2-[[2-[[(2S,3R)-2-amino-3-tert-butoxy-butanoyl]amino]-3-(2-fluorophenyl)-2-methyl-propanoyl]amino]-3-tert-butoxy-butanoyl]amino]-3-tert-butoxy-propanoyl]amino]-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[bis(4-docosoxyphenyl)methylcarbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG 1-7AA)

[0796] [ka] To a solution of Tag 1-6AA (6.5 g, 3.17 mmol, 1 eq.) and AA7 (6.30 g, 15.85 mmol, 5.0 eq.) in DCM (100 mL) was added DIC (1.80 g, 14.26 mmol, 2.21 mL, 4.5 eq.) and HOAt (1.94 g, 14.26 mmol, 2.00 mL, 4.5 eq.). The solution was then stirred at 20 °C for 2 h. TLC (PE / EtOAc = 1 / 1) showed that the reactant was completely consumed and a new spot was detected. To the reaction solution, ACN (400 mL) was added, but only a small amount of solid formed. HO (50 mL) was added, and the resulting suspension was filtered and then washed with ACN (200 mL × 2) to give an off-white solid. (Note: Filtration was very slow as the solid particles were very small.) N-Fmoc-tagged 1-7AA (6.6 g, 2.72 mmol, 85.69% yield) was obtained as a white solid, which was used directly in the next step. LCMS (ESI): RT = 5.437 min, C 74 H 89 FN 11 O 15 +Calculated mass 1390.46, observed m / z 1390.70 [M-tag-1+H] + ;(Tag1=C 57 H 99 O2, MS=815.76).

[0797] To a solution of N-Fmoc-tag 1-7AA (7.80 g, 3.21 mmol, 1 eq.) in THF (80 mL) was added piperidine (20 mL). The solution was then stirred at 20 °C for 2 h. TLC (PE / EtOAc = 1 / 1) showed that the reactant was completely consumed and a new spot was detected. The reaction mixture was added to ACN / HO = 1 / 8 (600 mL), and the resulting suspension was filtered. (Note: The filtration was very slow due to the very small solid particles.) The filter cake was triturated with THF / ACN = 1 / 12 (500 mL) and filtered. The filter cake was then washed with ACN (50 mL × 3) to obtain the crude product. Tag 1-7AA (5.0 g, 2.26 mmol, 70.54% yield) was obtained as a white solid, which was used directly in the next step. LCMS (ESI): RT = 3.550 min, C 45 H 62 N9O 10 + Calculated mass 888.45, observed m / z 888.50 [M-tag-1+H] + ;(Tag1=C 57 H 99 O2, MS=815.76).

[0798] Step 8: (3S)-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-carbamoyl-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-3-[[(2S)-2-[[(2S,3R)-2-[[3-(2-fluorophenyl)-2-[[(2S,3R)-3-hydroxy-2-[[2-[[(2 Synthesis of S)-2-[[3-[2-(1H-imidazol-5-yl)ethylamino]-2,2-dimethyl-3-oxo-propanoyl]amino]-3-(1H-tetrazol-5-yl)propanoyl]amino]acetyl]amino]butanoyl]amino]-2-methyl-propanoyl]amino]-3-hydroxy-butanoyl]amino]-3-hydroxy-propanoyl]amino]-4-oxo-butanoic acid (Tag1-M2)

[0799] [ka] To a solution of Tag 1-7AA (300 mg, 135.86 μmol, 1 eq.) and AA10-AA9-AA8-OH (116.11 mg, 142.66 μmol, 1.05 eq.) in DMF (0.5 mL) and DCM (1.5 mL), HATU (61.99 mg, 163.03 μmol, 1.2 eq.) was added. The mixture was then stirred at 20 °C for 2 h. TLC (PE / EtOAc = 1 / 1) showed that the reactant was completely consumed and a new spot was detected. The reaction mixture was quenched with water (50 mL) and diluted with DCM (80 mL). * 2). The combined organic layers were washed with brine (30 mL x 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give compound tag 1-10AA (408 mg, crude). LCMS (ESI): RT = 3.94 min, C 75 H 101 FN 20 O 17 Calculated m / z value of 786.375 [M-tag1+H] + ; m / z actual value 786.8 (tag 1 = C 57 H 99 O2, MS=815.76). LC-MS conditions: Method 7.

[0800] The crude tag 1-10AA was subjected to acidic cleavage using a TFA cocktail (10 mL, TFA:Tips:HO = 95:2.5:2.5) for 1.5 h. The reaction mixture was filtered, and the filtrate was concentrated to give a residue, which was purified by prep-HPLC (TFA; mobile phase: [water(TFA)-ACN] B%: 28%-58%, 12 min). M2 (40 mg, 25.45 μmol, 18.73% yield) was obtained as a white solid. LCMS: RT = 3.942 min, C 75 H 101 FN 20 O 17 Calculated m / z value: 786.375 [M+H] + m / z found 786.8. LC-MS conditions: Method 7.

[0801] HPLC: RT=9.66 min, purity 92.86%. HPLC method A: Column: YMC-Pack ODS-A 150 * 4.6 mm, 5 μm; 2.75 mL / 4 L TFA in water (solvent A) and 2.5 mL / 4 L TFA in ACN (solvent B) at a flow rate of 1.5 mL / min. Use an elution gradient of 10% to 80% (solvent B) for 10 min, hold at 80% for 5 min.

[0802] Synthesis of 2-[[(2S)-3-[1-[(2,4-dimethoxyphenyl)methyl]tetrazol-5-yl]-2-[[2,2-dimethyl-3-oxo-3-[2-(3-tritylimidazol-4-yl)ethylamino]propanoyl]amino]propanoyl]amino]acetic acid (AA10-AA9-AA8-OH)

[0803] [ka] To a solution of DMB-AA9 (3.23 g, 6.10 mmol, 2 eq) and DIEA (1.58 g, 12.20 mmol, 2.12 mL, 4 eq) in DMF (20 mL), HATU (2.32 g, 6.10 mmol, 2 eq) was added in one portion at 20 °C. The mixture was stirred at 20 °C for 0.2 hr. Compound 21 (2.82 g, 3.05 mmol, 37.94% purity, 1 eq) was added, and the mixture was stirred at 20 °C for 2 hr. After completion, the mixture was filtered, and the collected resin was washed with DMF (25 mL × 3) and DCM (25 mL × 3) to give the crude product Fmoc-22 (4.38 g, 3.05 mmol, 99.93% yield, 60% purity) on chlorotrityl chloride (CTC) resin as a light yellow solid. LCMS (ESI): RT = 0.942 min, C 30 H 30 Calculated m / z for N6O7Na: 609.22 [M+Na] + ; m / z observed value 609.1; LC-MS conditions: Method 13.

[0804] To a solution of Fmoc-22 (4.38 g, 3.05 mmol, 60% purity, 1 eq) in DMF (32 mL) was added piperidine (8 mL) in one portion at 20 °C. The mixture was stirred at 20 °C for 1 hr. The mixture was filtered and the collected resin was washed with DMF (25 mL × 3) and DCM (25 mL × 3) to give crude product 22 (3.7 g, 3.05 mmol, 99.96% yield, 52.7% purity) on CTC resin as a light yellow solid. LCMS (ESI): RT = 0.716 min, C 15 H 20 Calculated m / z value for N6O5Na: 387.15 [M+Na] + ; m / z observed value 387.1; LC-MS conditions: Method 13.

[0805] To a solution of AA10 (2.85 g, 6.09 mmol, 2 eq) and DIEA (1.57 g, 12.18 mmol, 2.12 mL, 4 eq) in DMF (20 mL) was added HATU (2.32 g, 6.09 mmol, 2 eq) in one portion at 20 °C. The mixture was stirred at 20 °C for 0.2 h. Compound 22 (3.7 g, 3.05 mmol, 52.7% purity, 1 eq) was added, and the mixture was stirred at 20 °C for 2 h. After completion, the reaction mixture was subjected to acidic cleavage using an AcOH cocktail (20 mL, AcOH:TFE:DCM = 1:1:8) for 2 h. The mixture was then filtered, and the collected resin was washed with DCM (5 mL × 3). The filtrate was washed with HO (10 mL × 3), and the combined aqueous layers were extracted with DCM (10 mL × 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue that was purified by C-18 reverse-phase chromatography (ISCO®; 40 g® C-18 Column, elution with a gradient of 0 to 60% ACN / HO (0.1% AcOH) at 40 mL / min, 1.2 L total volume, 30 min). After lyophilization, AA10-AA9-AA8-OH (260 mg, 287.51 μmol, 9.44% yield, 90% purity) was obtained as an off-white solid. LCMS (ESI): RT = 3.050 min, C 44 H 48 Calculated m / z for N9O7: 814.36 [M+H] + ; m / z observed value 814.4; LC-MS conditions: Method 13. [Example]

[0806] Synthesis of M2 to Tag2 (LPPS) 5.1 Tag2 Synthesis A synthetic route to make tag 2 is shown below in Scheme 3.

[0807] Scheme 3

[0808] [ka] Treatment of 2,4-dihydroxybenzaldehyde (Tag2-1) with 2.2 eq. of 1-bromodocosane in the presence of potassium carbonate in DMF / THF gave the bisC22-alkyl-substituted compound (Tag2-2). Grignard reaction of Tag2-2 with a Grignard reagent (Tag2-2a), followed by nucleophilic substitution of the alcohol (Tag2-3) with 9-fluorenylmethylcarbamate, gave Tag2-4. General Fmoc deprotection of Tag2-4 gave Tag2.

[0809] The alcohol tag (Tag 2-3) and the amine tag (Tag 2) are both new tags designed for use in the generation of short peptide acids and amides. The two hydrophobic C22 alkyl chains in Tag 2-3 and Tag 2 preserved the TCP hydrophobicity so that it could be precipitated in polar solvents during the synthesis process.

[0810] Step a: Synthesis of 2,4-di(docosoxy)benzaldehyde (Tag 2-2)

[0811] [ka] To a mixture of 2,4-dihydroxybenzaldehyde (Tag 2-1, 5 g, 36.20 mmol, 1 eq) and 1-bromodocosane (31.02 g, 79.64 mmol, 2.2 eq) in DMF (100 mL) and THF (20 mL), KCO (30.02 g, 217.20 mmol, 6 eq) was added and stirred at 90 °C for 12 h. Most of the reactant was consumed, and a new spot formed on TLC (PE / EtOAc = 5 / 1). The reaction mixture was poured into water (1000 mL), the mixture was stirred for 1 h, and then the precipitated solid was collected by filtration. The resulting solid was triturated with acetone (1000 mL), collected by filtration, and slurry washed in methanol. It was then dried under reduced pressure to give the desired compound, Tag 2-2 (27 g, crude material), as an off-white solid. 1H NMR (400MHz, chloroform-d) δ = 10.34 (s, 1H), 7.80 (d, J=8.8 Hz, 1H), 6.52 (dd, J=1.5, 8.8 Hz, 1H), 6.43 (d, J=1.9 Hz, 1H), 4.08 - 3.97 (m, 4H), 1.89 - 1.73 (m, 4H), 1.52 - 1.44 (m, 4H), 1.26 (s, 72H), 0.89 (t, J=6.7 Hz, 6H) ppm.

[0812] Step b: Synthesis of (2,4-bis(docosyloxy)phenyl)(4-methoxyphenyl)methanol (TAG 2-3)

[0813] [ka] To a solution of Tag 2-2 (31 g, 41.04 mmol, 1 eq) in THF (300 mL) was added bromo-(4-methoxyphenyl)magnesium (Tag 2-2a, 1 M, 123.13 mL, 3 eq), and the reaction mixture was stirred at 70 °C for 12 h. TLC (PE: EtOAc = 10:1) showed the reaction was complete. After cooling to room temperature, 1 N hydrochloric acid (100 mL) and THF (100 mL) were added to the reaction. The separated organic layer was washed with brine (50 mL × 2), dried over NaSO, filtered, and concentrated under reduced pressure to give a residue, which was triturated with ACN (150 mL) and MeOH (80 mL). The suspension was filtered and washed thoroughly with ACN (100 mL) and methanol (80 mL) to give the product tag 2-3 (36 g, 37.53 mmol, 91.43% yield, 90% purity) as a light yellow solid. 1H NMR (400MHz, chloroform-d) δ = 7.30 -7.27 (m, 2H), 7.17 - 7.00 (m, 1H), 6.90 - 6.77 (m, 2H), 6.49 - 6.32 (m, 2H), 5.98 - 5.49 (m, 1H), 3.99 - 3.85 (m, 4H), 3.85 - 3.70 (m, 4H), 3.53 - 3.35 (m, 2H), 1.80 - 1.68 (m, 4H), 1.43 (br d, J=5.0 Hz, 4H), 1.26 (s, 72H), 0.89 (t, J=6.7 Hz, 6H) ppm.

[0814] Step c: Synthesis of (9H-fluoren-9-yl)methyl((2,4-bis(docosyloxy)phenyl)(4-methoxyphenyl)methyl)carbamate (TAG 2-4)

[0815] [ka] To a solution of 9H-fluoren-9-ylmethylcarbamate (10.77 g, 45.03 mmol, 1.2 eq) in THF (300 mL) was added Tag 2-3 (36 g, 37.53 mmol, 90% purity, 1.0 eq) and MsOH (1.08 g, 11.26 mmol, 801.41 μL, 0.3 eq), and the reaction mixture was stirred at 70 °C for 12 h. TLC (PE: EtOAc = 10:1) showed the reaction was complete. After evaporation of most of the solvent, the residue was triturated with 5% aqueous NaHCO (10 mL), MeOH (80 mL), and ACN (150 mL). The suspension was filtered and washed thoroughly with ACN (120 mL) and methanol (80 mL) successively to give tag 2-4 (39 g, 32.36 mmol, 86.24% yield, 90% purity) as a light yellow solid. 1H NMR (400MHz, chloroform-d) δ =7.85 - 7.69 (m, 2H), 7.68 - 7.50 (m, 2H), 7.46 - 7.36 (m, 2H), 7.36 - 7.28 (m, 2H), 7.14 (br d, J=8.0 Hz, 2H), 6.81 (br d, J=8.3 Hz, 2H), 6.50 - 6.42 (m, 2H), 6.04 - 5.89 (m, 1H), 4.47 - 4.35 (m, 2H), 4.25 (br t, J=7.0 Hz, 1H), 3.97 - 3.91 (m, 2H), 3.86 - 3.71 (m, 5H), 3.42 - 3.19 (m, 1H), 1.80 - 1.64 (m, 4H), 1.48-1.36 (m, 4H), 1.1.32-1.1 (m, 72H), 0.95-0.82 (m, 6H) ppm.

[0816] Step d: Synthesis of (2,4-bis(docosyloxy)phenyl)(4-methoxyphenyl)methanamine (TAG 2)

[0817] [ka] To a solution of tag 2-4 (37 g, 30.70 mmol, 90% purity, 1 eq) in THF (240 mL) was added piperidine (51.73 g, 607.56 mmol, 60 mL, 19.79 eq), and the reaction mixture was stirred at 20 °C for 2 h. TLC (PE: EtOAc = 10:1) showed the reaction was complete. The mixture was added to ACN (800 mL), and the resulting suspension was filtered and washed thoroughly with ACN (200 mL) and methanol (100 mL) to give tag 2 (30 g, 29.57 mmol, 96.31% yield, 85% purity) as a light yellow solid. LC-MS (Method 7): RT = 2.038 min, m / z C 58 H 101 Calculated value of O3: 845.78 [M-NH3] + , Actual measured value 845.8. 1H NMR (400MHz, chloroform-d) δ =7.29 (d, J=8.5 Hz, 2H), 7.16 - 7.10 (m, 1H), 6.83 (d, J=8.8 Hz, 2H), 6.45 - 6.38 (m, 2H), 5.37 (s, 1H), 3.95 - 3.85 (m, 4H), 3.79 (s, 3H), 1.77 - 1.71 (m, 4H), 1.48 - 1.38 (m, 4H), 1.27 (s, 72H), 0.89 (s, 6H) ppm.

[0818] 5.2 Synthesis of Tag2-M2 Figure 3 and Table 5 outline the synthesis of M2 using Tag2.

[0819] [Table 6]

[0820] General Procedure C for One-Pot Amide Coupling and Fmoc Deprotection to Obtain Tag2-nAA To a solution of Tag2 or Tag2-(n-1)AA (1 eq.), AAn (1.5 eq.), and HOAt (1.5 eq.) in DCM (20 V) was added DIC (1.5 eq.) in one portion at 20°C. The mixture was stirred at 20°C for 12 hr. After completion, propan-1-amine (2 eq.) was added, and the mixture was stirred at 20°C for 0.5 hr. Then, 20% piperidine in THF (20 V) was added, and the mixture was stirred at 20°C for 1 hr. TLC (PE: EtOAc = 3:1) showed the reaction was complete. The mixture was diluted with ACN / MeOH = 3:1 (40 V). * 2) to afford the product Tag2-nAA as a light yellow solid, which was used directly in the next step without purification.

[0821] Step 1: Synthesis of (2S)-2-amino-N-((2,4-bis(docosyloxy)phenyl)(4-methoxyphenyl)methyl)-5-(3,5-dimethylphenyl)pentanamide (TAG2-1AA)

[0822] [ka] To a solution of Tag 2 (6.5 g, 7.16 mmol, 95% purity, 1 eq), AA1 (4.76 g, 10.74 mmol, 1.5 eq), and HOAt (1.46 g, 10.74 mmol, 1.50 mL, 1.5 eq) in DCM (80 mL) was added DIC (1.36 g, 10.74 mmol, 1.66 mL, 1.5 eq) in one portion at 20 °C. The mixture was stirred at 20 °C for 12 h. After completion, propan-1-amine (846.45 mg, 14.32 mmol, 1.18 mL, 2 eq) was added, and the mixture was stirred at 20 °C for 0.5 h. Then, piperidine (16.65 g, 195.52 mmol, 19.31 mL, 27.31 eq) in THF (80 mL) was added, and the mixture was stirred at 20 °C for 1 h. TLC (PE: EtOAc = 3:1) showed that the reaction was complete. The mixture was diluted with ACN / MeOH = 3:1 (150 mL * Trituration with 2) afforded tag 2-1AA (8.5 g, 6.78 mmol, 94.69% yield, 85% purity) as a light yellow solid, which was used directly in the next step without purification. LCMS (ESI): RT = 2.092 min, C 71 H 121 Calculated m / z for N2O4: 1065.92 [M+H] + ; m / z observed value 1065.9; LC-MS conditions: Method 7.

[0823] Step 2: Synthesis of (2S)-2-((S)-2-amino-3-(4'-(4-azidobutoxy)-2'-ethyl-[1,1'-biphenyl]-4-yl)propanamide)-N-((2,4-bis(docosyloxy)phenyl)(4-methoxyphenyl)methyl)-5-(3,5-dimethylphenyl)pentanamide (TAG2-2AA)

[0824] [ka] The reaction was carried out according to general procedure C for one-pot amide coupling and Fmoc deprotection using Tag 2-1AA (3 g, 2.39 mmol, 85% purity, 1 eq) and AA2 (2.18 g, 3.61 mmol, 1.51 eq). Crude Tag 2-2AA (3.6 g, 2.27 mmol, 94.68% yield, 90% purity) was obtained as a light yellow solid. LCMS (ESI): RT = 2.225 min, C 92 H 144 Calculated m / z for N6O6: 1430.11 [M+H] + ; m / z observed value 1431.0; LC-MS conditions: Method 7.

[0825] Step 3: Synthesis of (3S)-tert-butyl 3-amino-4-(((2S)-3-(4'-(4-azidobutoxy)-2'-ethyl-[1,1'-biphenyl]-4-yl)-1-(((2S)-1-(((2,4-bis(docosyloxy)phenyl)(4-methoxyphenyl)methyl)amino)-5-(3,5-dimethylphenyl)-1-oxopentan-2-yl)amino)-1-oxopropan-2-yl)amino)-4-oxobutanoate (TAG2-3AA)

[0826] [ka] The reaction was carried out according to general procedure C for one-pot amide coupling and Fmoc deprotection using Tag 2-2AA (3.8 g, 2.39 mmol, 90% purity, 1 eq) and AA3 (1.48 g, 3.59 mmol, 1.5 eq). Crude Tag 2-3AA (3.9 g, 2.19 mmol, 91.66% yield, 90% purity) was obtained as a light yellow solid. LCMS (ESI): RT = 3.660 min, C 38 H 50 Calculated m / z for N7O6: 700.37 [M-anchor-tBu+3H] + ;(Anchor=C 58 H 101 O3, MS = 845.8); m / z observed 700.4; LC-MS conditions: Method 7. (Anchor = Tag 2).

[0827] Step 4: Synthesis of (3S)-tert-butyl 3-((S)-2-amino-3-(tert-butoxy)propanamido)-4-(((2S)-3-(4'-(4-azidobutoxy)-2'-ethyl-[1,1'-biphenyl]-4-yl)-1-(((2S)-1-(((2,4-bis(docosyloxy)phenyl)(4-methoxyphenyl)methyl)amino)-5-(3,5-dimethylphenyl)-1-oxopentan-2-yl)amino)-1-oxopropan-2-yl)amino)-4-oxobutanoate (TAG2-4AA)

[0828] [ka] The reaction was carried out according to general procedure C for one-pot amide coupling and Fmoc deprotection using Tag 2-3AA (3.9 g, 2.19 mmol, 90% purity, 1 eq) and AA4 (1.26 g, 3.29 mmol, 1.5 eq). Crude Tag 2-4AA (3.8 g, 1.96 mmol, 89.44% yield, 90% purity) was obtained as a light yellow solid. LCMS (ESI): RT = 3.540 min, C 41 H 55 Calculated m / z for N8O8: 787.41 [M-anchor-2tBu+4H] + ;(Anchor=C 58 H 101 O3, MS = 845.8); m / z observed 787.5; LC-MS conditions: Method 7. (Anchor = Tag 2).

[0829] Step 5: Synthesis of (3S)-tert-butyl 3-((S)-2-(((2S,3R)-2-amino-3-(tert-butoxy)butanoyl)oxy)-3-(tert-butoxy)propanamido)-4-(((2S)-3-(4'-(4-azidobutoxy)-2'-ethyl-[1,1'-biphenyl]-4-yl)-1-(((2S)-1-(((2,4-bis(docosyloxy)phenyl)(4-methoxyphenyl)methyl)amino)-5-(3,5-dimethylphenyl)-1-oxopentan-2-yl)amino)-1-oxopropan-2-yl)amino)-4-oxobutanoate (TAG2-5AA)

[0830] [ka] The reaction was carried out according to general procedure C for one-pot amide coupling and Fmoc deprotection using Tag 2-4AA (3.8 g, 1.96 mmol, 90% purity, 1 eq) and AA5 (1.17 g, 2.94 mmol, 1.5 eq). Crude Tag 2-5AA (3.1 g, 1.55 mmol, 78.99% yield, 95% purity) was obtained as a light yellow solid. LCMS (ESI): RT = 3.510 min, C 45 H 62 N9O 10 m / z calculated value 888.45 [M-anchor-3tBu+5H] + ;(Anchor=C 58 H 101 O3, MS = 845.8); m / z observed 888.5; LC-MS conditions: Method 7. (Anchor = Tag 2).

[0831] Step 6: Synthesis of (3S)-tert-butyl 3-((S)-2-((2S,3R)-2-((S)-2-amino-3-(2-fluorophenyl)-2-methylpropanamido)-3-(tert-butoxy)butanamido)-3-(tert-butoxy)propanamido)-4-(((2S)-3-(4'-(4-azidobutoxy)-2'-ethyl-[1,1'-biphenyl]-4-yl)-1-(((2S)-1-(((2,4-bis(docosyloxy)phenyl)(4-methoxyphenyl)methyl)amino)-5-(3,5-dimethylphenyl)-1-oxopentan-2-yl)amino)-1-oxopropan-2-yl)amino)-4-oxobutanoate (TAG2-6AA)

[0832] [ka] The reaction was carried out according to general procedure C for one-pot amide coupling and Fmoc deprotection using Tag 2-5AA (1.5 g, 749.31 μmol, 95% purity, 1 eq) and AA6 (471.44 mg, 1.12 mmol, 1.5 eq). Tag 2-6AA (1.36 g, 588.20 μmol, 78.50% yield, 90% purity) was obtained as a light yellow solid. LCMS (ESI): RT = 3.726 min, C 55 H 72 FN 10 O 11 Calculated m / z value 1067.53 [M-anchor-3tBu+5H] + ;(Anchor=C 58 H 101 O3, MS = 845.8); m / z observed 1067.6; LC-MS conditions: Method 7. (Anchor = Tag 2).

[0833] Step 7: (4R,5S,8S,11S,14S,17S)-tert-butyl 5-amino-17-(((2S)-3-(4'-(4-azidobutoxy)-2'-ethyl-[1,1'-biphenyl]-4-yl)-1-(((2S)-1-(((2,4-bis(docosyloxy)phenyl)(4-methoxyphenyl)methyl)amino)-1-oxo-5-(m-tolyl)pentane Synthesis of (1-(2-yl)amino)-1-oxopropan-2-yl)carbamoyl)-11-(CAN-1-(tert-butoxy)ethyl)-14-(tert-butoxymethyl)-8-(2-fluorobenzyl)-2,2,4,8-tetramethyl-6,9,12,15-tetraoxo-3-oxa-7,10,13,16-tetraazanonadecane-19-oate (TAG2-7AA)

[0834] [ka] The reaction was carried out according to general procedure C for one-pot amide coupling and Fmoc deprotection using Tag 2-6AA (0.7 g, 302.75 μmol, 90% purity, 1 eq) and AA7 (360.99 mg, 908.25 μmol, 3 eq). Crude Tag 2-7AA (0.6 g, 241.27 μmol, 79.69% yield, 90% purity) was obtained as a light yellow solid. LCMS (ESI): RT = 3.977 min, C 59 H 79 FN 11 O 13 Calculated m / z value 1168.58 [M-anchor-4tBu+6H] + ;(Anchor=C 58 H 101 O3, MS=845.8); m / z found 1168.5; LC-MS conditions: Method 7 (Anchor=Tag 2) (filtration was very slow due to insufficient precipitation and collection of solid particles was difficult).

[0835] Step 8: (3S)-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-carbamoyl-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-3-[[(2S)-2-[[(2S,3R)-2-[[(2S)-3-(2-fluorophenyl)-2-[[(2S,3R)-3-hydroxy-2-[[2 Synthesis of -[[(2S)-2-[[3-[2-(1H-imidazol-5-yl)ethylamino]-2,2-dimethyl-3-oxo-propanoyl]amino]-3-(1H-tetrazol-5-yl)propanoyl]amino]acetyl]amino]butanoyl]amino]-2-methyl-propanoyl]amino]-3-hydroxy-butanoyl]amino]-3-hydroxy-propanoyl]amino]-4-oxo-butanoic acid (M2)

[0836] [ka] To a solution of the reactants AA10-AA9-AA8-OH (216.01 mg, 238.86 μmol, 90% purity, 1.1 eq) and Tag2-7AA (540 mg, 217.14 μmol, 90% purity, 1 eq) in DMF (2 mL) and DCM (6 mL) was added HATU (99.08 mg, 260.57 μmol, 1.2 eq) in one portion at 20 °C. The mixture was stirred at 20 °C for 2 h. LCMS showed complete conversion. The mixture was subjected to acidic cleavage using a TFA cocktail (14 mL, TFA:ethyl phenyl sulfide:HO = 95:2.5:2.5) for 50 min. The reaction mixture was filtered, and the filtrate was quenched with water / ACN = 4:1 (150 mL) and PE (20 mL). * The combined aqueous layers were lyophilized to give a residue, which was then purified by prep-HPLC (column: Welch Xtimate C18 100 * 40mm *Purification by HPLC using the following conditions (3 μm; mobile phase: [water (TFA)-ACN]; B%: 28% to 58%, 12 min) gave product M2 (50 mg, 31.58 μmol, 14.54% yield, 99.27% ​​purity) as a white solid. LCMS (ESI): RT = 3.910 min, C 75 H 101 FN 20 O 17 Calculated m / z value: 786.375 [M+H] + Found m / z 786.8; LC-MS conditions: Method 7. HPLC: RT = 4.86 min. HPLC conditions: Mobile phase: 1.0% ACN (0.1% TFA) in water to 5% ACN (0.1% TFA) in water in 1 min, then 5% ACN (0.1% TFA) in water to 100% ACN (0.1% TFA) in 5 min, hold at 100% ACN (0.1% TFA) for 2 min, return to 1.0% ACN (0.1% TFA) in water in 8.01 min, hold for 2 min. Flow rate: 1.2 ml / min. Column: Ultimate XB-C18, 3 um, 3.0 * 50mm. [Example]

[0837] Synthesis of M2 to Tag3 (LPPS) 6.1 Tag 3 Synthesis A synthetic route to generate tag 3 is shown in Scheme 4.

[0838] Scheme 4

[0839] [ka] Reagents and conditions: a) C 22 H 45Br (2.2 eq.), KCO (6 eq.), DMF / THF, 90 °C, 12 h, 99%; b) NHOH.HCl (3 eq.), EtN (5 eq.), toluene (20 mL), 100 °C, 12 h, 93%; c) DIBALH (3.0 eq.), toluene (30 mL), 100 °C, 12 h, 91%; d) 2-[4-[(2,4-dimethoxyphenyl)-(9H-fluoren-9-ylmethoxycarbonylamino)methyl]phenoxy]acetic acid (1.5 eq.), DIC (1.5 eq.), HOAt (1.5 eq.), DCM (2 mL), 20 °C, 12 h, 68%; e) 20% piperidine in THF, 20 °C, 2 h, 99%. Overall yield: 56.4%.

[0840] Bisalkylation of 2,4-dihydroxybenzaldehyde with C22-alkyl bromide in the presence of TEA in toluene, followed by treatment of tag 3-1 with an equal equivalent of hydroxylamine hydrochloride, afforded the oxime product tag 3-2. DIBAL reduction of tag 3-2 to the amine tag 3-3, followed by coupling with the acid tag 3-3a, afforded the amide tag 3-4. The reagent tag 3-3a (CAS: 145069-56-3, Rink amide linker), 4-[(2,4-dimethoxyphenyl)(fmoc-amino)methyl]phenoxyacetic acid, is commonly used in solid-phase synthesis for the preparation of C-terminal primary amide peptides. Finally, Fmoc deprotection of tag 3-4 afforded tag 3.

[0841] Step a: Synthesis of 2,4-di(docosoxy)benzaldehyde (TAG 3-1)

[0842] [ka] To a mixture of 2,4-dihydroxybenzaldehyde (2 g, 14.48 mmol, 1 eq) and 1-bromodocosane (12 g, 30.81 mmol, 2.13 eq) in DMF (50 mL) was added K2CO3 (12.01 g, 86.88 mmol, 6 eq), which was stirred at 90 °C for 12 h. Most of the reactant was consumed, and a new spot formed on TLC (PE / EtOAc = 5 / 1). After completion, the reaction mixture was poured into water (500 mL), the mixture was stirred for 1 h, and the precipitated solid was collected by filtration. The resulting solid was triturated with acetone (200 mL), and the solid was collected by filtration and slurry washed with methanol. After filtration, the resulting solid was dried under reduced pressure to give the title compound as an off-white solid. The resulting solid was triturated again with PE (100 mL), and the solid was collected by filtration. The desired compound tag 3-1 (10.9 g, 14.43 mmol, 99.67% yield, n / a purity) was obtained as an off-white solid. 1 H NMR (400MHz, chloroform-d) δ = 10.33 (s, 1H), 7.80 (d, J=8.6 Hz, 1H), 6.52 (d, J=8.8 Hz, 1H), 6.42 (d, J=2.1 Hz,1H), 4.08 - 3.97 (m, 4H), 1.89 - 1.69 (m, 4H), 1.52 - 1.42 (m, 4H), 1.26 (s, 72H), 0.89 (t, J=6.8 Hz, 6H) ppm.

[0843] Step b: Synthesis of (1E)-2,4-di(docosoxy)benzaldehyde oxime (TAG3-2)

[0844] [ka] To a mixture of tag 3-1 (6.5 g, 8.61 mmol, 1 eq) in toluene (90 mL), NHOH.HCl (5.98 g, 86.06 mmol, 10 eq) and EtN (8.71 g, 86.06 mmol, 11.98 mL, 10 eq) were added, and the reaction mixture was stirred at 100 °C for 12 h. Most of the reactant was consumed, and a new, more polar spot formed on TLC (PE / EtOAc = 10 / 1). After completion, the reaction mixture was cooled to 20 °C and poured into ACN (300 mL). The precipitated solid was then collected by filtration and slurry washed with methanol. The resulting solid was triturated with ACN (300 mL), and the solid was collected by filtration. After filtration, the resulting solid was dried under reduced pressure to give the title compound as a brown solid. The desired compound tag 3-2 (6.2 g, 8.05 mmol, 93.53% yield) was obtained as a brown solid. 1 H NMR (400MHz, chloroform-d) δ = 8.46 (s, 1H), 7.65 (d, J=8.5 Hz, 1H), 6.51 - 6.41 (m, 2H), 4.03 - 3.91 (m, 4H), 1.87 - 1.71 (m, 4H), 1.50 - 1.42 (m, 4H), 1.27 (s, 72H), 0.89 (t, J=6.8 Hz, 6H) ppm.

[0845] Step 3: Synthesis of [2,4-di(docosoxy)phenyl]methanamine (TAG3-3)

[0846] [ka] To a mixture of tag 3-2 (5 g, 6.49 mmol, 1 eq) in THF (50 mL) was added DIBALH (1 M, 19.47 mL, 3 eq) dropwise at 0 °C, and the reaction mixture was stirred at 90 °C for 12 h. Most of the reactant was consumed, and a small polar new spot formed on TLC (PE / EtOAc = 10 / 1). The reaction mixture was poured into saturated sodium tartrate tetrahydrate (100 mL), and the precipitated solid was collected by filtration and slurry washing with methanol. The resulting solid was triturated with ACN (30 mL), and the solid was collected by filtration. After filtration, the resulting solid was dried under reduced pressure to give the title compound tag 3-3 (4.5 g, 5.95 mmol, 91.66% yield) as an off-white solid. 1 H NMR (400MHz, chloroform-d) δ = 6.61 - 6.29 (m, 3H), 4.14 - 3.65 (m, 6H), 2.84 (br s, 2H), 1.83 - 1.73 (m, 4H), 1.4 - 1.5 (m, 4H), 1.27 (br s, 72H), 0.89 (br t, J=6.5 Hz, 6H) ppm.

[0847] Step 4: Synthesis of 9H-fluoren-9-ylmethyl N-[[4-[2-[[2,4-di(docosoxy)phenyl]methylamino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamate (Tag 3-4)

[0848] [ka] To a solution of tag 3-3 (2 g, 2.64 mmol, 1 eq) and 2-[4-[(2,4-dimethoxyphenyl)-(9H-fluoren-9-ylmethoxycarbonylamino)methyl]phenoxy]acetic acid tag 3-3a (2.14 g, 3.97 mmol, 1.5 eq) in DCM (20 mL) was added DIC (500.58 mg, 3.97 mmol, 614.21 μL, 1.5 eq) and HOAt (539.89 mg, 3.97 mmol, 554.87 μL, 1.5 eq), and the resulting mixture was stirred at 20 °C for 2 h. Most of the reactant was consumed, and a new, more polar spot formed on TLC (PE / EtOAc = 3 / 1). ACN (100 mL) was added to the reaction mixture, and the resulting suspension was filtered and washed with MeOH (30 mL × 2) to give an off-white solid. The resulting solid was triturated with ACN (30 mL) and collected by filtration. After filtration, the resulting solid was dried under reduced pressure to give the title compound tag 3-4 (2.3 g, 1.80 mmol, 68.06% yield) as an off-white solid. 1 H NMR (400MHz, chloroform-d) δ = 7.77 (br d, J=7.1 Hz, 2H), 7.60 (br d, J=6.8 Hz, 1H), 7.49 - 7.27 (m, 5H), 7.22 - 7.02 (m, 4H), 6.95 - 6.69 (m, 2H), 6.57 - 6.36 (m, 4H), 6.02 (br d, J=8.6 Hz, 1H), 5.75 (br d, J=7.6 Hz, 1H), 4.55 - 4.14 (m, 5H), 4.08 - 3.66 (m, 10H), 4.08 - 3.63 (m, 1H), 1.95 - 1.67 (m, 4H), 1.54 - 1.41 (m, 4H), 1.27 (s, 72H), 0.89 (t, J=6.8 Hz, 6H) ppm.

[0849] Step 5: Synthesis of 2-[4-[amino-(2,4-dimethoxyphenyl)methyl]phenoxy]-N-[[2,4-di(docosoxy)phenyl]methyl]acetamide (Tag 3)

[0850] [ka] To a solution of tag 3-4 (2.2 g, 1.72 mmol, 1 eq) in THF (20 mL) was added piperidine (6.90 g, 81.01 mmol, 8 mL, 47.05 eq), and the resulting mixture was stirred at 20 °C for 2 hr. Most of the reactant was consumed, and a new, large, polar spot formed on TLC (PE / EtOAc = 9 / 1). LCMS showed that the desired mass was detected. ACN (100 mL) was added to the reaction solution, and the resulting suspension was filtered and washed with MeOH (30 mL × 2) to give an off-white solid. The resulting solid was triturated with ACN (20 mL), and the solid was collected by filtration. The resulting solid was dried under reduced pressure to give the title compound tag 3 (1.8 g, 1.71 mmol, 99.04% yield) as an off-white solid. LCMS (ESI): RT = 2.055 min, C 68 H 114 Calculated mass of N2O6H: 1055.9 [M+H] + , C 68 H 112 No. 6 + 1038.8 [M-NH2] + , m / z measured value 1038.8 [M-NH2] + ; LCMS conditions: Method 12. 1 H NMR (400MHz, chloroform-d) δ = 7.22 (br d, J=8.5 Hz, 1H), 7.16 - 7.04 (m, 2H), 6.84 - 6.67 (m, 2H), 6.52 (d, J=2.3 Hz, 1H), 6.49 - 6.29 (m, 4H), 5.36 (s, 1H), 4.43 - 4.24 (m, 1H), 4.05 - 3.85 (m, 5H), 3.82 - 3.70 (m, 6H), 3.19 (s, 2H), 1.83 - 1.75 (m, 4H), 1.26 (s, 76H), 0.90 - 0.87 (m, 6H) ppm.

[0851] 6.2 Synthesis of Tag3-nAA Figure 4 and Table 6 below outline the synthesis of M2 using tag 3.

[0852] [Table 7]

[0853] General Procedure D for Amide Coupling to Generate N-Fmoc-Tagged 3-nAA To a solution of Tag3 or Tag3-(n-1)AA (1 eq.) and AAn (1.5 eq.) in DCM (20 V), DIC (1.5 eq.) and HOAt (1.5 eq.) were added, and the resulting mixture was stirred at 20 °C for 2 h. Most of the reactant was consumed, and a new, large, polar spot formed on TLC (PE / EtOAc = 3 / 1). A small sample was removed, and piperidine was added to the small sample to remove the Fmoc group. LCMS analysis of the sample indicated that the desired mass was detected. ACN (80 V) was added to the reaction solution, and the resulting suspension was filtered and washed with MeOH (30 V × 2) to give an off-white solid. The resulting solid was triturated with ACN (20 V), and the solid was collected by filtration. After filtration, the resulting solid was dried under reduced pressure to give the title compound N-Fmoc-Tag3-nAA as an off-white solid.

[0854] General Procedure E for Fmoc Deprotection to Generate Tag 3-nAA: A solution of N-Fmoc-tag 3-nAA (1 eq.) in 20% piperidine in THF (10 V) was stirred at 20 °C for 1 hour. Most of the reactant was consumed, and a new spot formed on TLC (PE / EtOAc = 9 / 1). LCMS showed that the desired mass was detected. ACN (100 V) was added to the reaction solution, and the resulting suspension was filtered and washed with MeOH (30 V × 2) to give the product. The resulting solid was triturated with ACN (20 V), and the solid was collected by filtration. The title compound, tag 3-nAA, was obtained as an off-white solid.

[0855] Step f: Synthesis of 9H-fluoren-9-ylmethyl N-[(1S)-1-[[[4-[2-[[2,4-di(docosoxy)phenyl]methylamino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]carbamate (N-Fmoc-tag 3-1AA)

[0856] [ka] The reaction was carried out using general procedure D for amide coupling with Tag 3 (1 g, 947.29 μmol, 1 eq) and AA1 (630.23 mg, 1.42 mmol, 1.5 eq) in DCM (20 mL). N-Fmoc-Tag 3-1AA (1.1 g, 742.66 μmol, 78.40% yield) was obtained as an off-white solid. LCMS (ESI): RT = 2.252 min, C 81 H 131 Calculated mass of N3O7H: 1259.9, measured m / z: 1260.0 [M-Fmoc+H] + ;LCMS conditions: Method 12. 1 H NMR (400MHz, chloroform-d) δ = 7.83 - 7.50 (m, 4H), 7.45 - 7.26 (m, 5H), 7.12 - 6.61 (m, 9H), 6.55 - 6.17 (m, 4H), 5.55 - 5.27 (m, 1H), 4.54 - 4.09 (m, 6H), 4.06 - 3.70 (m, 8H), 3.69 - 3.55 (m, 3H), 3.49 (br s, 6H), 3.17 (br s, 2H), 2.50 (br s, 2H), 2.27 (br s, 6H), 2.01 (s, 2H), 1.27 (br s, 76H), 0.91 - 0.87 (m, 6H) ppm.

[0857] Step g: Synthesis of (2S)-2-amino-N-[[4-[2-[[2,4-di(docosoxy)phenyl]methylamino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]-5-(3,5-dimethylphenyl)pentanamide (TAG3-1AA)

[0858] [ka] The reaction was carried out using N-Fmoc-tag 3-1AA (1 g, 675.15 μmol, 1 eq) in THF (10 mL) using general procedure E for Fmoc deprotection. Tag 3-1AA (0.81 g, 643.41 μmol, 95.30% yield) was obtained as an off-white solid. LCMS (ESI): RT = 6.848 min, C 81 H 131 Calculated mass of N3O7H: 1258.9 [M+H] + , m / z measured value 1259.0 [M+H] + ;LCMS conditions: Method 12.

[0859] Step h: Synthesis of 9H-fluoren-9-ylmethyl N-[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4-[2-[[2,4-di(docosoxy)phenyl]methylamino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]carbamate (N-Fmoc-tag 3-2AA)

[0860] [ka] The reaction was carried out using general procedure D for amide coupling with Tag 3-1AA (0.6 g, 476.60 μmol, 1 eq) and AA2 (432.30 mg, 714.90 μmol, 1.5 eq). N-Fmoc-Tag 3-2AA5 (0.56 g, 303.43 μmol, 63.66% yield) was obtained as an off-white solid. LCMS (ESI): RT = 6.048 min, C 49 H 54 Calculated mass of N6O5Na: 829.4, observed m / z: 829.4 [M-Rink amide-anchor + 2H] + ;Rink amide = C 17 H 18 O4, anchor = C 51 H 97 NO2; LC-MS conditions: Method 7. 1 H NMR (400MHz, Chloroform-d) δ = 7.75 (br d, J=6.3 Hz, 2H), 7.57 - 7.28 (m, 5H), 7.24 - 6.89 (m, 10H), 6.86 - 6.62 (m, 5H), 6.54 - 6.31 (m, 5H), 6.20 (br s, 1H), 5.27 (br s, 1H), 4.53 - 4.07 (m, 7H), 4.06 - 3.53 (m, 12H), 3.40 (br t, J=6.3 Hz, 2H), 3.17 (br d, J=7.3 Hz, 2H), 2.64 - 2.45 (m, 4H), 2.22 (br d, J=3.3 Hz, 6H), 1.95 - 1.71 (m, 8H), 1.27 (s, 82H), 1.06 (br t, J=7.2 Hz, 3H), 0.90 - 0.87 (m, 6H) ppm.

[0861] Step i: Synthesis of (2S)-2-((S)-2-amino-3-(4'-(4-azidobutoxy)-2'-ethyl-[1,1'-biphenyl]-4-yl)propanamide)-N-((4-(2-((2,4-bis(docosyloxy)benzyl)amino)-2-oxoethoxy)phenyl)(2,4-dimethoxyphenyl)methyl)-5-(3,5-dimethylphenyl)pentanamide (TAG3-2AA)

[0862] [ka] The reaction was carried out using N-Fmoc-tag 3-2AA (400 mg, 216.73 μmol, 1 eq) using general procedure E for Fmoc deprotection to give tag 3-2AA (380 mg, 210.67 μmol, 97.20% yield, 90% purity) as a light yellow solid. LC-MS conditions: Method 7.

[0863] Step j: Synthesis of (3S)-tert-butyl 3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-(((2S)-3-(4'-(4-azidobutoxy)-2'-ethyl-[1,1'-biphenyl]-4-yl)-1-(((2S)-1-(((4-(2-((2,4-bis(docosyloxy)benzyl)amino)-2-oxoethoxy)phenyl)(2,4-dimethoxyphenyl)methyl)amino)-5-(3,5-dimethylphenyl)-1-oxopentan-2-yl)amino)-1-oxopropan-2-yl)amino)-4-oxobutanoate (N-Fmoc-tagged 3-3AA)

[0864] [ka] The reaction was carried out using general procedure D for amide coupling with tag 3-2AA (380 mg, 210.67 μmol, 90% purity, 1 eq) and AA3 (130.02 mg, 316.01 μmol, 1.5 eq). N-Fmoc-tag 3-3AA (350 mg, 156.19 μmol, 74.14% yield, 90% purity) was obtained as a light yellow solid. LCMS (ESI): RT = 5.542 min, C 53 H 60 Calculated m / z for N7O8: 922.44 [M-Rink amide-anchor-tBu+3H] + ;Rink amide = C 17 H 18 O4, anchor = C 51 H 97 NO2; m / z found 922.3; LC-MS conditions: Method 7.

[0865] Step k: Synthesis of (3S)-tert-butyl 3-amino-4-(((2S)-3-(4'-(4-azidobutoxy)-2'-ethyl-[1,1'-biphenyl]-4-yl)-1-(((2S)-1-(((4-(2-((2,4-bis(docosyloxy)benzyl)amino)-2-oxoethoxy)phenyl)(2,4-dimethoxyphenyl)methyl)amino)-5-(3,5-dimethylphenyl)-1-oxopentan-2-yl)amino)-1-oxopropan-2-yl)amino)-4-oxobutanoate (TAG3-3AA)

[0866] [ka] The reaction was carried out using N-Fmoc-tag 3-3AA (350 mg, 156.19 μmol, 90% purity, 1 eq) using general procedure E for Fmoc deprotection. Tag 3-3AA (280 mg, 140.43 μmol, 89.91% yield, 90% purity) was obtained as a light yellow solid. LCMS (ESI): RT = 3.810 min, C 38 H 50 Calculated m / z for N7O6: 700.37 [M-Rink amide-anchor-tBu+3H] + ;Rink amide = C 17H 18 O4, anchor = C 51 H 97 NO2; m / z found 700.3; LC-MS conditions: Method 7.

[0867] Step 1: Synthesis of (3S)-tert-butyl 3-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(tert-butoxy)propanamido)-4-(((2S)-3-(4'-(4-azidobutoxy)-2'-ethyl-[1,1'-biphenyl]-4-yl)-1-(((2S)-1-(((4-(2-((2,4-bis(docosyloxy)benzyl)amino)-2-oxoethoxy)phenyl)(2,4-dimethoxyphenyl)methyl)amino)-5-(3,5-dimethylphenyl)-1-oxopentan-2-yl)amino)-1-oxopropan-2-yl)amino)-4-oxobutanoate (N-Fmoc-tag 3-4AA)

[0868] [ka] The reaction was carried out using general procedure D for amide coupling with tag 3-3AA (280 mg, 140.35 μmol, 90% purity, 1 eq) and AA4 (80.72 mg, 210.52 μmol, 1.5 eq). N-Fmoc-tag 3-4AA (300 mg, 124.94 μmol, 89.02% yield, 90% purity) was obtained as a light yellow solid. LCMS (ESI): RT = 5.337 min, C 56 H 65 N8O 10 Calculated m / z 1009.47 [M-Rink amide-anchor-2tBu+4H] + ;Rink amide = C 17 H 18 O4, anchor = C 51 H 97 NO2; m / z found 1009.4; LC-MS conditions: Method 7.

[0869] Step m: Synthesis of (3S)-tert-butyl 3-((S)-2-amino-3-(tert-butoxy)propanamido)-4-(((2S)-3-(4'-(4-azidobutoxy)-2'-ethyl-[1,1'-biphenyl]-4-yl)-1-(((2S)-1-(((4-(2-((2,4-bis(docosyloxy)benzyl)amino)-2-oxoethoxy)phenyl)(2,4-dimethoxyphenyl)methyl)amino)-5-(3,5-dimethylphenyl)-1-oxopentan-2-yl)amino)-1-oxopropan-2-yl)amino)-4-oxobutanoate (TAG 3-4AA)

[0870] [ka] The reaction was carried out using N-Fmoc-tag 3-4AA (280 mg, 116.67 μmol, 90% purity, 1 eq) using general procedure E for Fmoc deprotection. N-tag 3-4AA (230 mg, 106.83 μmol, 91.56% yield, 90% purity) was obtained as a light yellow solid. LC-MS conditions: Method 7.

[0871] Step n: (5S,8S,11S)-tert-butyl 11-(((2S)-3-(4'-(4-azidobutoxy)-2'-ethyl-[1,1'-biphenyl]-4-yl)-1-(((2S)-1-(((4-(2-((2,4-bis(docosyloxy)benzyl)amino)-2-oxoethoxy)phenyl)(2,4-dimethoxyphenyl)methyl)amino)-5-(3,5-di Synthesis of N-Fmoc-tagged 3-5AA (methylphenyl)-1-oxopentan-2-yl)amino)-1-oxopropan-2-yl)carbamoyl)-5-(CAN-1-(tert-butoxy)ethyl)-8-(tert-butoxymethyl)-1-(9H-fluoren-9-yl)-3,6,9-trioxo-2-oxa-4,7,10-triazatridecan-13-oate

[0872] [ka] To a solution of Tag 3-4AA (230 mg, 106.83 μmol, 90% purity, 1 eq), AA5 (63.69 mg, 160.24 μmol, 1.5 eq), and HOAt (21.81 mg, 160.24 μmol, 22.42 μL, 1.5 eq) in DCM (4 mL) was added DIC (20.22 mg, 160.24 μmol, 24.81 μL, 1.5 eq) in one portion under N at 20 °C. The mixture was stirred at 20 °C for 2 h. To monitor the reaction, a small amount of the residue was subjected to acidic cleavage using a TFA cocktail (0.1 mL, TFA:TIPS:HO:anisole = 95:2.5:2.5:4.5) for 1 h. LCMS showed that the reaction was completely converted. The mixture was diluted with ACN (20 mL). * 3) to give the crude product N-Fmoc-tagged 3-4AA (250 mg, 97.10 μmol, 90.90% yield, 90% purity) as a light yellow solid.

[0873] LCMS(ESI): RT=5.170 min, C 60 H 72 N9O 12 m / z calculated 1110.52 [M-Rink amide-anchor-3tBu+5H] + ;Rink amide = C 17 H 18 O4, anchor = C 51 H 97 NO2; m / z found 1110.4; LC-MS conditions: Method 7. [Example]

[0874] Synthesis of M2 to Tag4 (LPPS) 7.1 Tag 4 Synthesis A synthetic route for preparing tag 4 is shown below in Scheme 5.

[0875] Scheme 5

[0876] [ka] Reagents and conditions: a) C 22 H 45Br(3.3eq.), K2CO3(8eq.), DMF / THF, 90℃, 12h, 99.8%;b)LiAlH4(3eq.), THF, 70℃, 3h, 92 .3%;c)PBr3(3eq.), DCM, 20℃, 3h, 63%;d)55A(1eq.), K2CO3(6eq.), NMP, 90℃, 16h, 90.5% ;e)NaBH4(5eq.), EtOH(1V), THF(10V), 70℃, 12h, 89.9%;f)I2(1.8eq.), Li2CO3(0.02eq ), TMSN3(4.5eq.), DCM, 12h, 37℃, 80.9%;g)Pme3(3eq.), H2O, THF / DCM, 20℃, 12h, 66.8%. Total yield: 25.5%.

[0877] Tris-alkylation of the 3,4,5-trihydroxybenzoyl ester with C22-alkyl bromide, followed by LAH reduction of the ester tag 4-1, yielded the alcohol tag 4-2 (Matsuda et al., "Solution-Phase Synthesis of Oligodeoxyribonucleotides Using the H-Phosphonate Method with N-Unprotected 5'-Phosphite Monomers," RSC Adv. 11:38094-38107 (2021), incorporated herein by reference in its entirety). Conversion of the alcohol tag 4-2 to the bromo analog tag 4-3 with triphenylphosphine and carbon tetrabromide, followed by alkylation of tag 4-3a, generated the hexa-alkylated derivative tag 4-4. Conversion of the ketone tag 4-4 ​​to the alcohol tag 4-5 using sodium borohydride, followed by conversion of the alcohol tag 4-5 to the azide tag 4-6 using trimethylsilyl azide as the nucleophile in the presence of lithium carbonate as the base, and final reduction to the azide tag 4-6 using trimethylphosphine gave tag 4.

[0878] Step a: Synthesis of methyl 3,4,5-tri(docosoxy)benzoate (TAG 4-1)

[0879] [ka] To a mixture of methyl 3,4,5-trihydroxybenzoate (5 g, 27.15 mmol, 1 eq.) and 1-bromodocosane (34.90 g, 89.60 mmol, 3.3 eq.) in DMF (100 mL) and THF (20 mL), K2CO3 (30.02 g, 217.22 mmol, 8 eq.) was added, and the reaction mixture was stirred at 90 °C for 12 h. Most of the reactant was consumed, and a new, less polar spot formed on TLC (PE / EtOAc = 3 / 1). After the reaction was complete, the reaction mixture was poured into purified water (1000 mL), the mixture was stirred for 1 h, and the precipitated solid was collected by filtration. The resulting solid was triturated with acetone (300 mL), and the solid was collected by filtration and slurry washed with methanol. After filtration, the resulting solid was dried under reduced pressure to give the title compound as an off-white solid. Compound tag 4-1 (30.1 g, 27.12 mmol, 99.88% yield) was obtained as a gray solid. 1 H NMR (400MHz, chloroform-d) δ = 7.29 (s, 2H), 4.03 (dt, J=2.5, 6.4 Hz, 6H), 3.91 (s, 2H), 3.93 - 3.87 (m, 1H), 1.85 - 1.71 (m, 6H), 1.54 - 1.45 (m, 6H), 1.28 (s, 108H), 0.90 (t, J=6.8 Hz, 9H) ppm.

[0880] Step b: Synthesis of [3,4,5-tri(docosoxy)phenyl]methanol (TAG 4-2)

[0881] [ka] To a solution of compound tag 4-1 (1 g, 900.99 μmol, 1 eq.) in THF (10 mL) was added LiAlH (102.59 mg, 2.70 mmol, 2.70 mL, 3.0 eq.) at 0 °C. The mixture was then heated to 70 °C for 3 h. TLC (PE / EA = 20 / 1) showed that most of the reactants were completely consumed, and one new major spot was detected. The reaction mixture was quenched with 1N NCl, and the mixture was stirred at 20 °C for 1 h. It was then filtered, and the filter cake was concentrated to give the product. Compound tag 4-2 (0.9 g, 831.88 μmol, 92.33% yield) was obtained as a gray solid. 1 H NMR (400 MHz, chloroform-d) δ ppm 6.58 (s, 2 H) 4.62 (s, 2 H) 3.93 - 4.04 (m, 6 H) 1.72 - 1.87 (m, 6 H) 1.43 - 1.55 (m, 6 H) 1.28 (s, 108 H) 0.90 (t, J = 6.79 Hz, 9 H).

[0882] Step c: Synthesis of 5-(bromomethyl)-1,2,3-tri(docosoxy)benzene (TAG 4-3)

[0883] [ka] To a solution of compound tag 4-2 (0.9 g, 831.88 μmol, 1 eq.) in DCM (10 mL) was added a solution of PBr (675.54 mg, 2.50 mmol, 3.0 eq.) in DCM (10 mL). The solution was then stirred at 20 °C for 3 h. TLC (PE / EA = 10 / 1) showed that the reactant was completely consumed, with one small polar spot detected. The reaction mixture was quenched with HO (50 mL) and ACN (50 mL), and the mixture was stirred at 20 °C for 1 h. It was then filtered, and the filter cake was dried by evaporation of the solvent to give the product. Compound tag 4-3 (0.6 g, 524.12 μmol, 63.00% yield) was obtained as a white solid. 1H NMR (400 MHz, chloroform-d) δ ppm 6.58 (s, 2 H), 4.44 (s, 2 H), 3.92–4.00 (m, 6 H), 1.86 (dt, J=6.53, 3.26 Hz, 7 H), 1.42–1.52 (m, 8 H), 1.26 (s, 125 H), 0.86–0.93 (m, 12 H).

[0884] Step d: Synthesis of bis[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methanone (TAG 4-4)

[0885] [ka] A solution of compound tag 4-3a (500 mg, 2.33 mmol, 1.0 eq.) and K2CO3 (1.94 g, 14.00 mmol, 6.0 eq.) in NMP (150 mL) was degassed and purged with N2 10 times for approximately 10 min. The mixture was then heated to 90 °C, and a solution of tag 4-3 (5.37 g, 4.69 mmol, 2.01 eq.) in NMP (350 mL) was added under N2. The mixture was then degassed and purged with N2 10 times for approximately 10 min, and stirred at 90 °C for 16 hr. TLC (PE / EA = 20 / 1) showed that the starting material was completely consumed and the desired spot was detected. The reaction solution was slowly added to stirring ACN (1000 mL), and the reaction mixture was then filtered, and the filter cake was diluted with MeOH (250 mL). * 3) and ACN (250 mL * The filter cake was dried by evaporation of the residual solvent to give the product, Compound Tag 4-4 ​​(5.5 g, 2.11 mmol, 90.55% yield, 90% purity) as a white solid. 1H NMR (400 MHz, chloroform-d) δ ppm 7.80 (d, J=8.75 Hz, 4 H), 6.99 - 7.09 (m, 4 H), 6.63 (s, 4 H), 5.03 (s, 4 H), 3.95 - 4.01 (m, 12 H), 1.80 (br dd, J=14.07, 6.44 Hz, 12 H), 1.43 - 1.54 (m, 12 H), 1.26 (s, 226 H), 0.89 (t, J=6.69 Hz, 18 H).

[0886] Step e: Synthesis of bis[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methanol (TAG 4-5)

[0887] [ka] To a solution of tag 4-4 ​​(5.5 g, 2.35 mmol, 1 eq.) in THF (100 mL) and EtOH (25 mL), NaBH (444.21 mg, 11.74 mmol, 5.0 eq.) was slowly added, and the mixture was heated to 70 °C and stirred at 77 °C for 5 h. TLC (PE / EA = 10 / 1) showed that the starting material was completely consumed and a new spot was detected. The reaction mixture was slowly added to saturated NH Cl / ACN (550 mL / 750 mL). The slurry was filtered, and the resulting crystals were collected in ACN (250 mL). * 3) and methanol (250 mL * The product was obtained by washing with 3) Compound tag 4-5 (5.5 g, 2.11 mmol, 89.92% yield, 90% purity) as a white solid. 1H NMR (400 MHz, chloroform-d) δ ppm 7.30 (d, J=8.63 Hz, 4 H) 6.96 (d, J=8.63 Hz, 4 H) 6.62 (s, 4 H) 5.81 (br s, 1 H) 4.94 (s, 4 H) 3.95 - 4.01 (m, 12 H) 1.74 - 1.84 (m, 12 H) 1.48 (br s, 12 H) 1.28 (s, 226 H) 0.90 (t, J=6.75 Hz, 18 H).

[0888] Step f: Synthesis of 5-[[4-[azido-[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methyl]phenoxy]methyl]-1,2,3-tri(docosoxy)benzene (TAG 4-6)

[0889] [ka] To a mixture of compound Tag 4-5 (5.5 g, 2.35 mmol, 1 eq.) and I2 (1.07 g, 4.22 mmol, 850.79 uL, 1.8 eq.) in DCM (105 mL) was added Li2CO3 (3.47 mg, 46.93 umol, 31.67 uL, 0.02 eq.) and TMSN3 (1.22 g, 10.56 mmol, 1.39 mL, 4.5 eq.), and the reaction mixture was stirred at 37 °C for 12 h. The reactant was consumed and the desired spot formed on TLC (PE / EtOAc = 20 / 1). The reaction solution was slowly added to stirring ACN (750 mL) and saturated Na2SO3 (250 mL). The reaction mixture was then filtered, and the filter cake was dissolved in methanol (250 mL). * 3) and ACN (250 mL * The filter cake was dried by evaporation of the residual solvent to give the product, tag 4-6 (5.0 g, 1.90 mmol, 80.95% yield, 90% purity) as a white solid. 1H NMR (400 MHz, chloroform-d) δ ppm 7.23 (br d, J=8.63 Hz, 4H) 6.96 (br d, J=8.63 Hz, 4H) 6.61 (s, 4 H) 5.62 (s, 1H) 5.65 (s, 1H) 4.93 (s, 4H) 3.94 - 3.99 (m, 12H) 1.76 - 1.84 (m, 12H) 1.43 - 1.50 (m, 12H) 1.27 (s, 226H) 0.87 - 0.90 (m, 18H).

[0890] Step g: Synthesis of bis[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methanamine (TAG 4)

[0891] [ka] To a solution of tag 4-6 (4.5 g, 1.90 mmol, 1 eq.) in THF (50 mL), DCM (50 mL), and HO (2.5 mL) was added PMe3 (1 M, 5.70 mL, 3.0 eq.). The reaction mixture was then stirred at 20 °C for 12 h. TLC (PE / EA = 20 / 1) showed that the reactant was completely consumed, and TLC (PE / EA = 1 / 1) showed that the desired spot was detected. The reaction solution was slowly added to stirring ACN (800 ml) and HO (200 mL). The reaction mixture was then filtered, and the filter cake was triturated again with ACN (800 mL), and then the filter cake was dissolved in MeOH (250 mL). * 3) and ACN (250 mL * 3) The filter cake was dried by evaporation of residual solvent to give the product Tag 4 (3.5 g, 1.27 mmol, 66.84% yield, 85% purity) as a white solid. 1H NMR (400 MHz, Chloroform-d) δ ppm 7.28 - 7.33 (m, 4H) 6.90 - 6.99 (m, 4H) 6.60 - 6.66 (m, 4H) 5.16 (s, 1H) 4.93 (s, 4H) 3.95 - 4.01 (m, 12H) 1.76 - 1.82 (m, 12H) 1.48 (br s, 12H) 1.28 (s, 226H) 0.88 - 0.92 (m, 18H).

[0892] 7.2 Synthesis of Tag4-M2 Figure 5 and Table 7 below outline the synthesis of M2 using tag 4.

[0893] [Table 8]

[0894] General Procedure F for Amide Coupling to Prepare N-Fmoc-Tagged 4-nAA To a solution of Tag4 or Tag4-(n-1)AA (1 eq.), AAn (1.5 eq.), and HOAt (1.5 eq.) in DCM (20 V) was added DIC (1.5 eq.) and HOAt (1.5 eq.). The solution was stirred at 20 °C for 12 h. A small sample was removed and subjected to acidic cleavage using a TFA cocktail (TFA:TIPS:HO = 95:2.5:2.5). The sample was filtered and analyzed by LCMS, which showed that the reactant was consumed and the desired mass was detected. ACN (80 V) was added to the reaction solution, and the resulting suspension was filtered and washed with ACN (20 V × 2) to give an off-white solid. The solvent was removed by filtration, and the filter cake was dried by evaporation of the residual solvent to give the product N-Fmoc-Tag4-nAA as an off-white solid, which was used directly in the next step.

[0895] General Procedure G for Fmoc Deprotection to Obtain Tag 4-nAA A solution of compound N-Fmoc-tag 4-nAA (1 eq.) in 20% piperidine (20 V) in THF and DCM was stirred at 20 °C for 2 h. TLC (PE / EA = 1 / 1) showed that the reactant was completely consumed and the desired spot was detected. ACN (200 V) was added to the reaction solution, and the resulting suspension was filtered and washed with ACN (40 V × 2) to obtain tag 4-nAA as an off-white solid. The crude product was dried under reduced pressure and used directly in the next step without further purification.

[0896] Step 1: Synthesis of (2S)-2-amino-N-[bis[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methyl]-5-(3,5-dimethylphenyl)pentanamide (TAG4-1AA)

[0897] [ka] The reaction was carried out according to general procedure F for amide coupling using Tag 4 (2.5 g, 1.07 mmol, 1 eq.) and AA1 (709.88 mg, 1.60 mmol, 1.5 eq.). N-Fmoc-Tag 4-1AA (2.6 g, 751.31 μmol, 70.41% yield, 80% purity) was obtained as an off-white solid. LC-MS: RT = 4.061 min, C 28 H 31 N2O3 + Calculated mass 443.23, measured m / z 443.20 [M-anchor + H] + ;(Anchor=C 159 H 286 O8, MS=2325.21). LCMS conditions: Method 12. Anchor=Tag 4.

[0898] The reaction was carried out according to general procedure G for Fmoc deprotection using N-Fmoc-tag 4-1AA (2.0 g, 722.41 umol, 1 eq.). Tag 4-1AA (1.6 g, crude) was obtained as a white solid and used directly in the next step. LC-MS: 443.2 (M+H). + .

[0899] Step 2: Synthesis of (2R)-2-[[(2S)-2-amino-3-[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]propanoyl]amino]-N-[bis[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methyl]-5-(3,5-dimethylphenyl)pentanamide (TAG 4-2AA)

[0900] [ka] The reaction was carried out according to general procedure F for amide coupling using tag 4-1AA (1.6 g, 628.37 μmol, 1 eq.) to give N-Fmoc-tag 4-2AA (1.9 g, 424.52 μmol, 67.56% yield, 70% purity) as a white solid, which was used directly in the next step. LC-MS: RT=5.525 min, C 49 H 55 N6O5 + Calculated mass 807.42, measured m / z 807.40 [M-anchor + H] + ;(Anchor=C 159 H 286 O8, MS=2325.21). LCMS conditions: Method 12. (Anchor=Tag 4).

[0901] The reaction was carried out according to general procedure G for Fmoc deprotection using N-Fmoc-tag 4-2AA (1.9 g, 606.46 μmol, 1 eq.). Tag 4-2AA (1.7 g, 584.05 μmol, 96.31% yield) was obtained as an off-white solid. LC-MS: RT=3.534 min, C 47 H 55 N6O5 + Calculated mass 783.42, measured m / z 783.40 [M-anchor + C 13 H 11 O2 + H] + ;(Anchor=C 159 H 286 O8, MS=2325.21);C 34 H 45 N6O3 +Calculated mass 585.35, measured m / z 585.40 [M-anchor + H] + ;(Anchor=C 159 H 286 O8, MS=2325.21); LCMS conditions: Method 12. (Anchor=Tag 4).

[0902] Step 3: Synthesis of tert-butyl (3S)-3-amino-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[bis[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methylcarbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG 4-3AA)

[0903] [ka] The reaction was carried out according to general procedure F for amide coupling using tag 4-2AA (1.7, 584.05 μmol, 1 eq.). N-Fmoc-tag 4-3AA (1.9 g, 345.02 μmol, 59.07% yield, 60% purity) was obtained as an off-white solid. LC-MS: RT=5.159 min, C 53 H 60 N7O8 + Calculated mass 922.41, measured m / z 922.40 [M-anchor + H] + ;(Anchor=C 159 H 286 O8, MS=2325.21). LCMS conditions: Method 12. (Anchor=Tag 4).

[0904] The reaction was carried out according to general procedure G for Fmoc deprotection using N-Fmc-tag 4-3AA (1.9 g, 575.04 μmol, 1 eq.) to give tag 4-3AA (1.50 g, 486.71 μmol, 84.64% yield) as an off-white solid. LC-MS: RT=3.633 min, C 51 H 60 N7O8 +Calculated mass 898.44, observed m / z 898.50 [M-tBu-anchor + C 13 H 11 O2 + H] + ;(Anchor=C 159 H 286 O8, MS=2325.21);C 38 H 50 N7O6 + Calculated mass 700.37, observed m / z 700.50 [M-tBu-anchor + H] + ;(Anchor=C 159 H 286 O8, MS=2325.21); LCMS conditions: Method 12. (Anchor=Tag 4).

[0905] Step 4: Synthesis of tert-butyl (3S)-3-[[(2S)-2-amino-3-tert-butoxy-propanoyl]amino]-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[bis[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methylcarbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG 4-4AA)

[0906] [ka] The reaction was carried out according to general procedure F for amide coupling using tag 4-3AA (1.5 g, 486.71 μmol, 1 eq.) in DCM (50 mL). N-Fmoc-tag 4-4AA (1.45 g, 420.62 μmol, 86.42% yield) was obtained as an off-white solid. LC-MS: RT=5.019 min, C 56 H 65 N8O 10 + Calculated mass 1009.40, measured m / z 1009.40 [M-anchor + H] + ;(Anchor=C 159 H 286 O8, MS=2325.21). LCMS conditions: Method 12. (Anchor=Tag 4).

[0907] The reaction was carried out according to general procedure G for Fmoc deprotection using N-Fmoc-tag 4-4AA (1.4 g, 406.11 μmol, 1 eq.). Tag 4-4AA (1.30 g, 403.09 μmol, 99.26% yield) was obtained as an off-white solid. LC-MS: RT=3.527 min, C 54 H 65 N8O 10 + Calculated mass 985.47, observed m / z 985.50 [M-tBu-anchor + H] + ;(Anchor=C 159 H 286 O8, MS=2325.21);C 41 H 55 N8O8 + Calculated mass 787.41, observed m / z 787.50 [M-tBu-anchor + H] + ;(Anchor=C 159 H 286 O8, MS=2325.21); LCMS conditions: Method 12. (Anchor=Tag 4).

[0908] Step 5: Synthesis of tert-butyl (3S)-3-[[(2S)-2-[[(2S,3R)-2-amino-3-tert-butoxy-butanoyl]amino]-3-tert-butoxy-propanoyl]amino]-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[bis[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methylcarbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG 4-5AA)

[0909] [ka] The reaction was carried out according to general procedure F for amide coupling using tag 4-4AA (1.45 g, 420.62 μmol). N-Fmoc-tag 4-5AA (1.3 g, 360.66 μmol, 85.74% yield) was obtained as an off-white solid. LC-MS: RT=4.894 min, C 60 H 72 N9O 12 + Calculated mass 1110.52, observed m / z 1110.60 [M-tBu-anchor + H] + ;(Anchor=C 159 H 286 O8, MS=2325.21);C 73 H 82 N9O 14 + Calculated mass 1308.57, observed m / z 1308.60 [M-tBu-anchor + H] + ;(Anchor=C 159 H 286 O8, MS=2325.21); LCMS conditions: Method 12. (Anchor=Tag 4).

[0910] The reaction was carried out according to general procedure G for Fmoc deprotection using N-Fmoc-tag 4-5AA (1.30 g, 360.66 μmol, 1 eq.). Tag 4-5AA (1.0 g, 295.66 μmol, 81.98% yield) was obtained as an off-white solid, which was used directly in the next step. LC-MS: RT=3.557 min, C 58 H 72 N9O 12 + Calculated mass 1086.52, observed m / z 1086.30 [M-3tBu-anchor + C 13 H 11 O2+4H] + ;(Anchor=C 159 H 286 O8, MS=2325.21);C 45 H 62 N9O 10 + Calculated mass 888.45, observed m / z 888.50 [M-3tBu-anchor + 4H]+; (anchor = C 159 H286 O8, MS=2325.21); LCMS conditions: Method 12. (Anchor=Tag 4).

[0911] Step 6: Synthesis of tert-butyl (3S)-3-[[(2S)-2-[[(2S,3R)-2-[[2-amino-3-(2-fluorophenyl)-2-methyl-propanoyl]amino]-3-tert-butoxy-butanoyl]amino]-3-tert-butoxy-propanoyl]amino]-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[bis[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methylcarbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG 4-6AA)

[0912] [ka] The reaction was carried out according to general procedure F for amide coupling using Tag 4-5AA (1.0 g, 295.66 μmol, 1 eq.). N-Fmoc-Tag 4-6AA (0.8 g, 211.43 μmol, 71.51% yield) was obtained as an off-white solid. LC-MS: RT=5.208 min, C 70 H 82 FN 10 O 13 + Calculated mass 1289.60, observed m / z 1289.60 [M-3tBu-anchor + 4H] + ;(Anchor=C 159 H 286 O8, MS=2325.21); LCMS conditions: Method 12. (Anchor=Tag 4).

[0913] The reaction was carried out according to general procedure G for Fmoc deprotection using N-Fmoc-tag 4-6AA (0.8 g, 211.43 μmol, 1 eq.). Tag 4-6AA (0.7 g, 196.55 μmol, 92.96% yield) was obtained as an off-white solid. LC-MS: RT=3.697 min, C 68 H 82 FN 10 O 13 + Calculated mass 1265.60, measured m / z 1265.70 [M-3tBu-anchor + C 13 H 11 O2+4H] + ;(Anchor=C 159 H 286 O8, MS=2325.21); RT=3.630 min, C 55 H 72 FN 10 O 11 + Calculated mass 1067.53, observed m / z 1067.60 [M-3tBu-anchor + 4H] + ;(Anchor=C 159 H 286 O8, MS=2325.21); LCMS conditions: Method 12. (Anchor=Tag 4).

[0914] Step 7: tert-Butyl (3S)-3-[[(2S)-2-[[(2S,3R)-2-[[2-[[(2S,3R)-2-amino-3-tert-butoxy-butanoyl]amino]-3-(2-fluorophenyl)-2-methyl-propanoyl]amino]-3-tert-butoxy-butanoyl]amino]-3-tert-butoxy-propanoyl]amino]-4-[[( Synthesis of 1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[bis[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methylcarbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG 4-7AA)

[0915] [ka] The reaction was carried out according to general procedure F for amide coupling using Tag 4-6AA (0.7 g, 196.55 μmol, 1 eq.). N-Fmoc-Tag 4-7AA (0.7 g, 177.62 μmol, 90.37% yield) was obtained as an off-white solid. LC-MS: RT=5.224 min, C 74 H 89 FN 11 O 15 + Calculated mass 1390.64, observed m / z 1390.60 [M-3tBu-anchor + 4H] + ;(Anchor=C 159 H 286 O8, MS=2325.21); LCMS conditions: Method 12. (Anchor=Tag 4).

[0916] The reaction was carried out according to general procedure G for Fmoc deprotection using N-Fmoc-tag 4-7AA (0.7 g, 177.62 μmol, 1 eq.). Tag 4-7AA (0.58 g, 155.97 μmol, 87.81% yield) was obtained as an off-white solid, which was used directly in the next step. LC-MS: RT=3.869 min, C 72 H 89 FN 11 O 15 + Calculated mass 1366.65, measured m / z 1366.70 [M-3tBu-anchor + C 13 H 11 O2+4H] + ;(Anchor=C 159 H 286 O8, MS=2325.21);C 59 H 79 FN 11 O 13 + Calculated mass 1168.58, observed m / z 1168.60 [M-3tBu-anchor + 4H] + ;(Anchor=C 159 H 286 O8, MS=2325.21); LCMS conditions: Method 12. (Anchor=Tag 4).

[0917] Step 8: tert-Butyl (3S)-3-[[(2S)-2-[[(2S,3R)-2-[[2-[[(2S,3R)-2-[(2-aminoacetyl)amino]-3-tert-butoxy-butanoyl]amino]-3-(2-fluorophenyl)-2-methyl-propanoyl]amino]-3-tert-butoxy-butanoyl]amino]-3-tert-butoxy-propanoyl]amino Synthesis of ]-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[bis[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methylcarbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG 4-8AA)

[0918] [ka] The reaction was carried out according to general procedure F for amide coupling using Tag 4-7AA (0.6 g, 161.35 μmol, 1 eq.). N-Fmoc-Tag 4-8AA (0.56 g, 140.07 μmol, 86.81% yield) was obtained as an off-white solid. LC-MS: RT=5.092 min, C 76 H 92 FN 12 O 16 + Calculated mass of 1447.67, [M-3tBu-anchor+4H] + (Anchor = C159H286O8, MS = 2325.21); LCMS conditions: Method 12. (Anchor = Tag 4).

[0919] The reaction was carried out according to general procedure G for Fmoc deprotection using N-Fmoc-tag 4-8AA (0.56 g, 140.07 μmol, 1 eq.). Tag 4-8AA (0.5 g, 132.42 μmol) was obtained as an off-white solid and used directly in the next step. LC-MS: RT=3.848 min, C 61 H 82 FN 12 O 14+ Calculated mass 1225.60, observed m / z 1225.70 [M-3tBu-anchor + 4H] + ;(Anchor=C 159 H 286 O8, MS=2325.21);C 74 H 92 FN 12 O 16 + Calculated mass 1423.67, measured m / z 1423.70 [M-3tBu-anchor + C 13 H 11 O2+4H] + ;(Anchor=C 159 H 286 O8, MS=2325.21); LCMS conditions: Method 12. (Anchor=Tag 4).

[0920] Step 9: tert-Butyl (3S)-3-[[(2S)-2-[[(2S,3R)-2-[[2-[[(2S,3R)-2-[[2-[[(2S)-2-amino-3-[1-[(2,4-dimethoxyphenyl)methyl]tetrazol-5-yl]propanoyl]amino]acetyl]amino]-3-tert-butoxy-butanoyl]amino]-3-(2-fluorophenyl)-2-methyl-propanoyl]amino]-3-tert-butoxy-butanoyl Synthesis of [[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[bis[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methylcarbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG 4-9AA)

[0921] [ka] The reaction was carried out according to general procedure F for amide coupling using Tag 4-8AA (105.19 mg, 198.64 μmol, 1.5 eq.), DMB-AA9 (105.19 mg, 198.64 μmol, 1.5 eq.), DIC (25.07 mg, 198.64 μmol, 30.76 μL, 1.5 eq.), and HOAt (27.04 mg, 198.64 μmol, 27.79 μL, 1.5 eq.). N-Fmoc-Tag 4-9AA (0.5 g, 116.62 μmol, 88.07% yield) was obtained as an off-white solid. LC-MS: RT = 4.817 min, C 80 H 98 FN 17 O 17 + Calculated mass 793.86, measured mass 794.30 [M-3tBu-PMB-Anchor+7H] 2+ ;(Anchor=C 159 H 286 O8, MS=2325.21);C 95 H 106 FN 17 O 19 Calculated mass 892.89, measured mass 893.50 [M-3tBu-PMB-Anchor+7H] + ;(Anchor=C 159 H 286 O8, MS=2325.21); LCMS conditions: Method 12. (Anchor=Tag 4).

[0922] The reaction was carried out according to general procedure G for Fmoc deprotection using N-Fmoc-tag 4-9AA (0.5 g, 116.62 μmol, 1 eq). Tag 4-9AA (0.5 g, crude) was obtained as an off-white solid. LC-MS: RT=3.838 min, C 64 H 86 FN 17 O 16 + Calculated mass 683.80, observed m / z 683.20 [M-4tBu-DMB-anchor + 7H] 2+ ;(Anchor=C 159 H 286 O8, MS=2325.21);C 77 H 96 FN17 O 18 2+ Calculated mass 782.85, observed m / z 782.30 [M-4tBu-DMB-anchor + 7H] + ;(Anchor=C 159 H 286 O8, MS=2325.21); LCMS conditions: Method 12. (Anchor=Tag 4).

[0923] Step 10: tert-butyl(3S)-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[bis[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methylcarbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-3-[[(2S)-3-tert-butoxy-2-[[(2S,3R)-3-tert-butoxy-2-[[2-[[(2S,3R)-3 Synthesis of tert-butoxy-2-[[2-[[(2S)-3-[1-[(2,4-dimethoxyphenyl)methyl]tetrazol-5-yl]-2-[[2,2-dimethyl-3-oxo-3-[2-(3-tritylimidazol-4-yl)ethylamino]propanoyl]amino]propanoyl]amino]acetyl]amino]butanoyl]amino]-3-(2-fluorophenyl)-2-methyl-propanoyl]amino]butanoyl]amino]propanoyl]amino]-4-oxo-butanoate (TAG 4-10AA)

[0924] [ka] The reaction was carried out according to general procedure F for amide coupling using Tag 4-9AA (0.5 g, 123.00 μmol, 1 eq.), AA10 (86.26 mg, 184.50 μmol, 1.5 eq.), DIC (23.28 mg, 184.50 μmol, 28.57 μL, 1.5 eq.), and HOAt (25.11 mg, 184.50 μmol, 25.81 μL, 1.5 eq.) in DCM (15 mL) and THF (15 mL). Tag 4-10AA (0.5 g, 110.75 μmol, 90.04% yield) was obtained as an off-white solid, which was used directly in the next step. LC-MS: RT = 3.874 min, C 75 H 101 FN 20 O 172 + Calculated mass 786.37, measured mass 787.00 [M-4tBu-DMB-Trt-Anchor+8H] 2+ ;(Anchor=C 159 H 286 O8, MS=2325.21);C 88 H 111 FN 20 O 19 Calculated mass 885.41, measured mass 886.00 [M-4tBu-DMB-Trt-Anchor + 8H] + ;(Anchor=C 159 H 286 O8, MS=2325.21); LCMS conditions: Method 12. Note: (Anchor = Tag 4).

[0925] Step 11: Synthesis of bis[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methanamine (M2)

[0926] [ka] To a mixture of Tag 4-10AA (432.77 mg, 95.86 μmol, 1 eq.) in TFA (4.5 mL) and HO (0.25 mL) was added triisopropylsilane (143.92 mg, 908.82 μmol, 186.67 μL, 9.48 eq.) in one portion under N at 20 °C. The mixture was stirred at 20 °C for 1.5 hr. LCMS showed that the desired mass was detected. LC-MS: RT = 3.822 min, C 75 H 100 FN 20 O 17 m / z calculated value 1570.75 [M-Boc-4tBu+2H] 2+ m / z found 786.20; LCMS conditions: Method 12. The mixture was filtered and the filtrate was diluted with t-BuOMe (1000 mL) to give a precipitate, which was centrifuged for 10 min (5000 R) to give the crude product. The residue was purified by Prep-HPLC (column: Welch Xtimate C18 100 * 40mm * The resulting mixture was purified by HPLC using a HPLC-MS / MS column (HPLC: 3 μm; mobile phase: [water (TFA)-ACN]; B%: 30% to 60%, 15 min). Compound M2 (1 mg, 5.98 e-1 μmol, purity 94%) was obtained as a white solid. LCMS (ESI): RT = 3.748 min, C 75 H 100 FN 20 O 17 m / z calculated value 1570.75 [M-Boc-4tBu+2H] 2+ ; m / z found 786.20; LCMS conditions: Method 12. HPLC RT=9.411 min, purity 94%. [Example]

[0927] Synthesis of M1 and M3 to Tag5 (LPPS) 8.1 Tag 5 Synthesis Tag 5 was prepared in 10 steps as shown in Scheme 6 below.

[0928] Scheme 6

[0929] [ka] Tag 5-3 was prepared from methyl 3,4,5-trihydroxybenzoate as described by Horenstein et al., "Synthesis of Unprotected (±)-Tunichrome An-1, a Tunicate Blood Pigment," J. Am. Chem. Soc. 111(16):6242-6246 (1989), which is incorporated herein by reference in its entirety. Reductive amination of tag 5-3a with 2 eq. of tag 5-3 gave tag 5-4 in 70% yield. Removal of the N-allyl group of tag 5-4 using 1,3-dimethylbarbituric acid in the presence of Pd(PPh3)4 catalyst provided tag 5-5, which was coupled with tag 5-5a to form tag 5-6. Fmoc deprotection of tag 5-6 in piperidine gave tag 5-7. N-Alloc protection of tag 5-7, followed by removal of TBS from tag 5-8 and alkylation with C22-alkyl bromide in one pot produced tag 5-9. Removal of the N-alloc group from tag 5-9 afforded the final product, tag 5. Further improvement was attempted by replacing the N-Fmoc of tag 5-5a with N-alloc, but tag 5-5a without N-Fmoc was poorly soluble in organic solvents. The amounts and yields of each step are summarized in Table 8 below.

[0930] [Table 9-1]

[0931] [Table 9-2]

[0932] Step a: Synthesis of methyl 3,4,5-tris((tert-butyldimethylsilyl)oxy)benzoate (TAG5-1)

[0933] [ka] To a clear solution of methyl 3,4,5-trihydroxybenzoate (3 kg, 16.29 mol, 1 eq) in DMF (24 L), imidazole (4.99 kg, 73.31 mol, 4.5 eq) was added in one portion to obtain a clear solution. TBDMSCl (8.59 kg, 57.02 mol, 6.99 L, 3.5 eq) was added in four portions (2.148 kg x 4) over 2 h under N2 at a rate sufficient to maintain the internal temperature between 25 °C and 30 °C. After addition, a yellow solution was obtained. The reaction was stirred at 25 °C under N2 for 16 h until a thick suspension formed. TLC showed complete consumption of the starting material and one new major spot (petroleum ether: EtOAc = 9:1, Rf = 0.58) was detected. The reaction mixture was transferred to a transit barrel (2 * The mixture was pumped into a 50-L jacketed flask (20 L) and saturated NaHCO3 (15 L) solution was poured into the flask. 20 L (1 / 2 volume) of the reaction suspension was added to the above saturated NaHCO3 (15 L) solution with vigorous stirring and extracted with PE (6 L × 2). The other (1 / 2 volume) reaction mixture was treated as above. All organic layers were combined, washed with brine (10 L), and dried over Na2SO4. The organic layer was filtered and concentrated under reduced pressure (approximately 45 °C, -0.8 atm) to give approximately 10 kg of a pale green oil. The pale green oil was dissolved in MeOH (28 L) and stirred at 0–4 °C (internal temperature) for 12 h. A white solid precipitated. The product, Tag 5-1 (7.8 kg, 14.80 mol, 90.86% yield), was obtained as a white solid via filtration and drying under reduced pressure. 1 HNMR: (400 MHz, CDCl3) δ = 7.23 (s, 2H), 3.85 (s, 3H), 1.00 (s, 9H), 0.99 - 0.86 (m, 18H), 0.24 (s, 12H), 0.15 (s, 6H); LC-MS: (analyzed by method 1) Rt= 3.183 min, m / z=527.3 [M+H] + .

[0934] Step b: Synthesis of [3,4,5-tris[[tert-butyl(dimethyl)silyl]oxy]phenyl]methanol (TAG 5-2)

[0935] [ka] A solution of tag 5-1 (4 kg, 7.59 mol, 1 eq) in THF (15 L) was added to a suspension of LAH (216.09 g, 5.69 mol, 0.75 eq) in THF (16 L) at a rate sufficient to maintain a gentle reflux (internal temperature 66 °C). The reaction mixture was stirred at an internal temperature of 66 °C (oil bath temperature: 77 °C) for 3 h. TLC showed complete consumption of tag 5-1 and the formation of one new spot (petroleum ether:ethyl acetate = 10:1, Rf = 0.41). The reaction mixture was cooled (internal temperature: 0 °C) and diluted with 12 L of methyl tert-butyl ether (MTBE). The reaction mixture was rapidly stirred, and 216 mL of water was added dropwise until the color changed from gray to white. Then, 216 mL of NaOH solution (wt%, 15%) was added. The resulting mixture was stirred for 10 min. Water (620 mL) and silica gel (500 g) were added sequentially to the reaction mixture. The heterogeneous mixture was filtered through Celite, and the resulting filtrate was concentrated under vacuum to give a colorless oil. This was further evacuated with an oil pump to remove the solvent residue, affording tag 5-2 (3.48 kg, 6.84 mol, 90.1% yield, 98% purity) as a white solid. 1 LC-MS: (Analyzed by method 1) Rt= 3.25 min, m / z=481.3 [M+H-H2O] + .

[0936] Step c: Synthesis of 3,4,5-tris[[tert-butyl(dimethyl)silyl]oxy]benzaldehyde (TAG 5-3)

[0937] [ka] To a suspension of PCC (1.58 kg, 7.32 mol, 1.05 eq) in DCM (12 L) was added a solution of tag 5-2 (3.48 kg, 6.98 mol, 1 eq) in DCM (16 L) at 0 °C. The reaction mixture was stirred at 20 °C for 20 h. TLC showed that the reactant tag 5-2 was completely consumed, with one new major spot (petroleum ether:ethyl acetate = 10:1, PMA, Rf = 0.61) detected. MTBE (12 L) and silica gel (800 g) were added sequentially to the reaction mixture, which was then stirred for 10 min and filtered through Celite. The filtrate was concentrated under vacuum, and the resulting dark brown oil was diluted with petroleum ether (15 L), passed through a short pad of silica / Celite (800 g / 400 g), and concentrated to give tag 5-3 (3.13 kg, 6.30 mol, 90.34% yield) as a pale yellow solid. 1 H NMR (400 MHz, CDCl3) δ = 9.74 (s, 1H), 7.04 (s, 2H), 1.00 (s, 9H), 0.96 (s,18H), 0.26 (s, 12H), 0.16 (s, 6H). LC-MS: (analyzed by method 1) Rt= 3.550 minutes, m / z = 497.4 [M+H] + Tag5-3 was used directly in the next step without purification.

[0938] Step d: Synthesis of N,N-bis[[3,4,5-tris[[tert-butyl(dimethyl)silyl]oxy]phenyl]methyl]prop-2-en-1-amine (TAG 5-4)

[0939] [ka] To a solution of tag 5-3a (240 g, 2.51 mol, 98% purity, 1 eq) was added NaOAc (721.82 g, 8.80 mol, 3.5 eq) in THF (8 L) and MeOH (12.5 L). The mixture was stirred under N for 30 min, then a solution of tag 5-3 (1.40 kg, 2.77 mol, 98% purity, 1.1 eq) in THF (1.5 L) and HOAc (3.02 kg, 50.28 mol, 2.88 L, 20 eq) were added sequentially at 20 °C. The reaction mixture was stirred for 30 min. A solution of NaBHCN (157.98 g, 2.51 mol, 1 eq) in THF (1.5 L) was added dropwise to the reaction mixture over 30 min, and the reaction was then stirred at 25 °C for 30 min. An additional solution of tag 5-3 (1.66 kg, 3.27 mol, 98% purity, 1.3 eq) in THF (1.5 L) was added, and the reaction was stirred for 30 min. An additional solution of NaBHCN (236.98 g, 3.77 mol, 1.5 eq) in THF (1.5 L) was added dropwise to the reaction mixture over 1 h, and the reaction was then stirred at 25 °C for 12 h. TLC showed that the reactant tag 5-3 was consumed, with one major spot appearing (petroleum ether / ethyl acetate = 20 / 1, staining reagent: PMA, Rf = 0.66). Two impurities, tag 5-2 and tag 5-4', were also detected. The reaction was quenched by the addition of saturated aq. NaHCO (18 L, 3.6 kg NaHCO suspended in 16 L HO), and the separated aq. solution was then dissolved in MTBE (12 L). * The crude material was mixed with silica gel (4.5 kg) and purified by column chromatography using petroleum ether / ethyl acetate (1 / 0 to 100 / 10) to give tag 5-4 (1.88 kg, 1.73 mol, 71.2% yield based on 3a). 1H NMR (400 MHz, CDCl3) δ = 6.50 (s, 4H), 6.02 - 5.71 (m, 1H), 5.21 (d, J = 17.6 Hz, 1H), 5.12 (d, J = 10.0 Hz, 1H), 3.37 (br s, 4H), 2.98 (br d. + .

[0940] Step e: Synthesis of 1-[3,4,5-tris[[tert-butyl(dimethyl)silyl]oxy]phenyl]-N-[[3,4,5-tris[[tert-butyl(dimethyl)silyl]oxy]phenyl]methyl]methanamine (TAG 5-5)

[0941] [ka] To a solution of 1,3-dimethylbarbituric acid (CAS No.: 769-42-6, 885.13 g, 5.67 mol, 3.2 eq) in DCM (12.5 L) was added a solution of tag 5-4 (1.9 kg, 1.77 mol, 95% purity, 1 eq) in DCM (4 L). The reaction was degassed and purged with N three times, then Pd(PPh3)4 (61.41 g, 53.15 mmol, 0.03 eq) was added. The reaction mixture was degassed and purged with N2 three times and stirred under N2 atmosphere at an internal temperature of 39 °C (oil bath temperature: 45 °C) for 4 h. TLC showed complete consumption of the reactant tag 5-4 and the formation of one new spot (petroleum ether:ethyl acetate = 10:1, NH3.HO (0.5%, v / v), ninhydrin, Rf = 0.62). The reaction mixture was cooled to 10 °C and quenched by the addition of NH3.HO (400 mL, wt%: 25%). A white solid precipitated. Sulfhydryl silica gel (350 g), silica gel (700 g), and Na2SO4 (1000 g) were added to the reaction, and the reaction mixture was stirred for 30 min. The reaction mixture was filtered through a plug of Celite (500 g) to give tag 5-5 (1.69 kg, 1.64 mol, 92.6% yield, 95% purity) as a yellow oil, which was dissolved in DCM (10 L) and used directly without purification. 1 LC-MS: (Analyzed using method 1) Rt = 4.21 min, m / z=978.5 [M+H] + .

[0942] Step f: Synthesis of 9H-fluoren-9-ylmethyl N-[[4-[2-[bis[[3,4,5-tris[[tert-butyl(dimethyl)silyl]oxy]phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamate (Tags 5-6)

[0943] [ka] To a solution of tag 5-5a (913.17 g, 1.69 mol, 1.13 eq) in DCM (6 L) was added HOAt (245.71 g, 1.81 mol, 252.53 mL, 1.2 eq) and DIC (227.82 g, 1.81 mol, 279.53 mL, 1.2 eq), and the reaction mixture was stirred at 25 °C for 30 min. Then, a solution of tag 5-5 (1.55 kg, 1.50 mol, 95% purity, 1 eq) in DCM (10 L) was added, and the reaction mixture was stirred at 25 °C for 8 hr. TLC showed that the reactant tag 5-5 was consumed, and one new major spot (petroleum ether:ethyl acetate = 5:1, 4% DCM for improved solubility, Rf = 0.39) was detected. The reaction mixture was diluted with saturated NaHCO3 (5 L) and HCl (10 L). * 3) and then DCM (3 L) * The resulting mixture was extracted with 2). The combined organic layers were washed with brine (2.5 L), dried over NaSO, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO, petroleum ether / ethyl acetate = 20 / 1 to 1 / 1) to give tag 5-6 as a white solid (2.2 kg, 1.25 mol, yield 82.85%, purity 85%). 1H NMR (400 MHz, CDCl3) δ (ppm) = 7.77 (br d, J = 7.3 Hz, 2H), 7.59 (br d, J = 6.4 Hz, 2H), 7.39 (br d, J = 6.6 Hz, 2H), 7.31 (br d, J = 6.4 Hz, 2H), 7.24 - 7.14 (m, 1H), 7.11 - 7.08 (m, 2H), 6.79 (br d, J = 8.5 Hz, 2H), 6.50 - 6.47 (m, 2H), 6.38 (m, 2H), 6.31 (s, 2H), 6.06 - 5.99 (m, 1H), 5.86 - 5.72 (m, 1H), 4.66 (s, 2H), 4.46 - 4.38 (m, 4H), 4.29 - 4.19 (m, 3H), 3.81 (s, 3H), 3.74 (s, 3H), 1.00 (s, 18H), 0.92 (s, 36H), 0.19 (s, 24H), 0.13 (s, 12H). LC-MS: (analyzed by method 1) Rt= 5.39 min, m / z=251.0 [M+6H] 6+ .

[0944] Step g: Synthesis of 2-[4-[amino-(2,4-dimethoxyphenyl)methyl]phenoxy]-N,N-bis[[3,4,5-tris[[tert-butyl(dimethyl)silyl]oxy]phenyl]methyl]acetamide (TAG 5-7)

[0945] [ka] To tag 5-6 (2.2 kg, 1.25 mol, 85% purity, 1 eq) in THF (16 L) was added piperidine (530.63 g, 6.23 mol, 615.44 mL, 5 eq). The reaction mixture was degassed and purged with N2 three times, and then the mixture was stirred under N2 atmosphere at 60 °C for 2 hr. TLC showed the formation of one new major spot (petroleum ether:ethyl acetate = 1:1, Rf = 0.16). The reaction mixture was concentrated to give the crude material (1.8 kg, 1.18 mol, 94.91% yield, 84% purity) as a yellow oil, which was dissolved in DCM (12 L) and saturated ammonium chloride (10 L). * The residue was washed with 2), water (10 L), and brine (10 L). It was then dried over NaSO and filtered to give a solution which was concentrated to give tag 5-7 (1.8 kg with 84% purity, 94.9% yield). Tag 5-7 was used directly in the next step without purification. 1 NMR (400 MHz, CDCl3) δ (ppm) 7.27 (br d, J = 8.6 Hz, 2H), 7.12 (d, J = 8.0 Hz, 1H), 6.81 (d, J = 8.6 Hz, 2H), 6.44 (s, 2H), 6.39 (s, 2H), 6.31 (s, 2H), 5.38 (s, 1H), 4.66 (s, 2H), 4.43 (s, 2H), 4.29 (s, 2H), 3.79 - 3.76 (m, 6H), 1.00 (s,18H), 0.92 (s, 36H), 0.25 - 0.16 (m, 24H), 0.13 (s, 12H). LC-MS: (analyzed by method 1) Rt= 4.30 min, m / z=1260.7 [M+H-NH3] + .

[0946] Step h: Synthesis of allyl N-[[4-[2-[bis[[3,4,5-tris[[tert-butyl(dimethyl)silyl]oxy]phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamate (TAGs 5-8)

[0947] [ka] To a solution of tag 5-7 (1.8 kg, 1.18 mol, 84% purity, 1 eq) in DCM (20 L) was added DIEA (305.78 g, 2.37 mol, 412.10 mL, 2 eq) and allyl carbonochloride (285.17 g, 2.37 mol, 250.15 mL, 2 eq). The reaction mixture was stirred at 25 °C for 12 h. TLC showed that the starting material was completely consumed and one new major spot (petroleum ether:ethyl acetate = 5:1, Rf = 0.45) was detected. The reaction mixture was quenched by the addition of saturated NH4Cl (5 L), followed by DCM (5 L). * The combined organic layer was extracted with saturated NH4Cl (5 L * The crude product was purified by column chromatography (SiO, petroleum ether / ethyl acetate = 20:1 to 3:1, 2% DCM was added to the elution solvent to promote solubility) to give tags 5-8 in two fractions totaling 1.59 kg (97% yield with >98% purity). Fraction 1 (1.35 kg, 991.02 mmol, 99% purity) gave a yellow oil in 83.78% yield. Fraction 2 (380 g, approximately 80% purity) was further purified to give 236 g, 160.02 mmol, 98.1% purity as a pale yellow oil. 1H NMR (400 MHz, CDCl3) δ (ppm) = 7.10 (br d, J = 8.8 Hz, 3H,26,28,37), 6.77 (d, J = 8.6 Hz, 2H), 6.52 - 6.42 (m, 2H), 6.37 (s, 2H), 6.30 (s, 2H), 6.01 - 5.78 (m, 3H), 5.30 (br dd, J = 1.6, 17.2 Hz, 1H), 5.21 (br dd, J = 1.3, 10.4 Hz, 1H), 4.64 (s, 2H), 4.61 - 4.58 (m, 2H), 4.42 (s, 2H), 4.26 (s, 2H), 3.80 (s, 3H), 3.71 (s, 3H), 0.99 (s, 18H), 0.95 - 0.88 (m, 36H), 0.18 (s, 24H), 0.12 (s, 12H). LC-MS: (Method 2) Rt = 2.22 min, m / z=1379.74 [M+H3O] + m / z=2741.2 [2M+HO] +

[0948] Step i: Synthesis of allyl N-[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamate (TAG 5-9)

[0949] [ka] Five batches of reactions were run in parallel on a 50 g scale. To a mixture of tag 5-8 (50.0 g, 33.80 mmol, 1 eq) and 1-bromodocosane (86.2 g, 206.19 mmol, 6.1 eq) was added K2CO3 (42.1 g, 304.2 mmol, 9 eq), 18-crown-6 (5.4 g, 20.3 mmol, 0.6 eq), and KF (17.67 g, 304.21 mmol, 9 eq). The reaction mixture was degassed and purged with N2 three times, and then the mixture was stirred at 85 °C under a N2 atmosphere for 24 h. LCMS showed that tag 5-8 had been consumed. TLC showed one new major spot (THF / petroleum ether = 1:5, Rf = 0.35). The reaction mixture was suspended in THF (400 mL) and filtered through a silica gel plug to remove inorganic salts. The filtrate was concentrated to give a residue, which was purified by flash silica gel chromatography (ISCO®; 330 g SepaFlash® Silica Flash Column, 0–50% THF / petroleum ether gradient elution: 120 mL / min) to give tag 5-9 (76 g, 30.06 mmol, 88.94% yield).

[0950] Four additional reactions were carried out and purified as above. The five batches of purified tag 5-9 were then combined and triturated with MeOH (200 mL, 70 °C, 2 hr), then cooled to 20 °C and filtered to give a total of 386 g of tag 5-9 (purity: approximately 97.7%). 1H NMR (400 MHz, CDCl3) δ (ppm) = 7.12 (br d, J = 8.5 Hz, 3H), 6.81 (br d, J = 8.6 Hz, 2H), 6.50 - 6.43 (m, 2H), 6.40 (s, 2H), 6.30 (s, 2H), 6.00 (br d, J = 8.8 Hz,1H), 5.97 - 5.87 (m, 1H), 5.86 - 5.72 (br d, J = 8.6 Hz, 1H), 5.38 - 5.26 (m, 1H), 5.25 -5.08 (m, 1H), 4.71 (s, 2H), 4.59 (br d, J = 5.4 Hz, 2H), 4.50 (s, 2H), 4.40 (s, 2H), 3.93 (br t, J = 5.7 Hz, 4H), 3.90 - 3.81 (m, 8H), 3.80 (s, 3H), 3.72 (s, 3H), 1.82 - 1.68 (m, 12H), 1.53 - 1.20 (m, 228H), 0.89 (t, J = 6.8 Hz). LC-MS (Method 3): Rt= 11.33 min, m / z=2544.7 / 2545.99 [M+H3O] + NMR data were consistent with the target structure using HSQC. The chemical shift of C55 was 69.30 ppm, and HMBC:H55 correlated to C13.

[0951] Step j: Synthesis of 2-[4-[amino-(2,4-dimethoxyphenyl)methyl]phenoxy]-N,N-bis[[3,4,5-tri(docosoxy)phenyl]methyl]acetamide (TAG 5)

[0952] [ka] To a solution of 1,3-dimethylhexahydropyrimidine-2,4,6-trione (15.3 g, 97.7 mmol, 3.2 eq) in DCM (560 mL) was added tag 5-9 (80 g, 30.54 mmol, 1 eq). The reaction was degassed and purged with N three times, then Pd(PPh3)4 (1.41 g, 1.22 mmol, 0.04 eq) was added. The mixture was degassed, purged with N three times, and stirred under N2 atmosphere at 45 °C for 6 h. TLC showed that tag 5-9 was consumed and one new major spot (petroleum ether:THF = 1:1, Rf = 0.14, PMA) was detected. The reaction was concentrated, and the residue was triturated with MeOH (1 L) at 70 °C for 1 h, then cooled to 20 °C. The mixture was filtered to yield the crude product, which was further triturated with ACN (1 L, 70 °C, 1 hr), cooled to 20 °C, filtered, washed sequentially with ACN (400 mL), MeOH (400 mL), EtOAc (400 mL), and petroleum ether (400 mL), and then dried under vacuum to give Tag 5 (74.8 g, 29.32 mmol, 95.99% yield, 95.79% purity) as an off-white solid. Four additional replicate reactions were carried out as above to produce all Tag 5 (303.9 g at 95.5% purity). 1 H NMR (400 MHz, CDCl3) δ (ppm) 7.14 - 7.25 (m, 2 H), 7.03 (br d, J=7.75 Hz, 1 H), 6.76 (br d, J=8.25 Hz, 2 H), 6.34 (br d, J=12.38 Hz, 4 H), 6.22 (br s, 2 H), 5.30 (br s, 1 H), 4.65 (s, 2 H), 4.23 - 4.50 (m, 4 H), 3.63 - 3.96 (m, 18 H), 1.67 (br d, J=5.50 Hz, 12 H), 1.37 (br s, 12 H), 1.18 (s, 216 H), 0.81 (br t, J=6.44 Hz, 18 H) ppm. 13C NMR (101 MHz, CDCl3) δ = 168.42, 159.83, 157.67, 156.74, 153.62, 153.27, 137.65 - 137.58, 137.58 - 137.50, 131.79, 131.09, 128.17, 128.07, 114.29, 107.04, 105.04, 104.00, 98.74, 77.25, 73.48, 73.43, 69.18, 69.14, 67.75, 55.37, 55.34, 52.88, 49.67, 48.50, 31.97, 30.42, 29.77, 29.71, 29.57, 29.51, 29.42, 26.23, 22.74, 14.16. LC-MS (Method 3): Rt = 11.20 min, m / z=2444.02 [M+H] + .

[0953] 8.2 Characterization of Tag5 The characterization of Tag5 is shown in Table 9.

[0954] [ka]

[0955] [Table 10]

[0956] IR Data The powder sample of Tag 5 was placed directly on the surface of the ATR, while the sample completely covered the crystal (ZnSe) surface of the ATR. The sample was compressed using a sample compression bar to ensure even distribution and complete contact with the crystal surface of the ATR. The IR conditions used to collect the IR data are listed in Table 10.

[0957] [Table 11]

[0958] λ=2915.54cm -1indicates a CH(CH3) bond, λ=2848.94 cm -1 indicates a CH(CH2) bond, λ=1647.99 cm - 1 shows a C=O (C=ONR2) bond, λ = 1505.99 cm -1 indicates a C-N bond, λ=1466.52 cm -1 indicates a C=C(Ph) bond, λ=1379.34 cm -1 indicates a CH(CH3) bond, λ=1174.13 cm -1 showed COC binding.

[0959] Determination of moisture content The sample was weighed and placed in a clean, dry vial. The vial was sealed. After the instrument reached the set temperature (150°C), the vial was placed in the oven. After the Karl Fischer reaction was complete, the moisture content of the sample was obtained (Table 11). The moisture content was determined to be 0.26%.

[0960] [Table 12]

[0961] Ion chromatography The chromatographic conditions used are shown in Tables 12-13. The following ion amounts were determined: Cl - :0.01%;Br - :0.01%;F - :<0.01;CO3 2- :0.02%.

[0962] [Table 13]

[0963] [Table 14]

[0964] Differential scanning calorimeter The sample (1-8 mg) was placed in a high-pressure crucible and sealed with an alignment tool. The prepared sample was placed in the corresponding position of the instrument, the software parameters were selected, and the sequence was started (Table 14).

[0965] The first peak (melting point, endothermic) was observed at 76.3° C. The second peak (C22 must be volatile, endothermic) was observed at approximately 371° C.

[0966] [Table 15]

[0967] Residual Solvent (RS) Determination Residual solvents (GC and HS conditions) are shown in Table 15. Instrument: SHIMADZU-GC2030 & HS-20. The column used for chromatography was DB-624 (25 m x 0.2 mm, ID 1.12 μm). P / N: 128-1324. Split ratio: 30. Control mode: linear velocity. Flow rate: 1.2 mL / min. Carrier gas: N2. Detector: flame ionization detector (FID). All solvent residues were found to be below the limit of quantitation (LOQ).

[0968] [Table 16]

[0969] Solubility Test Tag 5 (10 mg) was added to solvent (100 μL) and shaken at different temperatures (Table 16). Tag 5 was found to be soluble in DCM and THF, soluble in any ratio of PE / DCM and PE / THF, insoluble in DMF and DMF / DCM, and insoluble in DMSO and DMSO / DCM (see Table 16).

[0970] [Table 17]

[0971] Stability testing A stability test for Tag 5 was performed at pH=2 for 2 hours using the following procedure. Tag 5 (1 g, 98.01% purity) was added to a 50 mL flask equipped with a magnetic stir bar in THF (20 mL), and the mixture was stirred for 5 minutes to dissolve the material. A solution of TFA in DCM (v / v=1:5, 2 mL) was added dropwise to the solution containing Tag 5 to bring the pH to 2, and the resulting solution was stirred continuously for 2 hours. The reaction was monitored by LCMS, which showed more than five peaks. It was concluded that Tag 5 was not very stable in acidic solutions (pH=2).

[0972] 8.3 Synthesis of GLP1 Peptide Mimetics to Tag 5 (LPPS) The synthesis of a GLP1 peptidomimetic (M1) to tag 5 is shown in Figures 6 and 10A-B and in Table 17. The amino acids used in the preparation of this peptidomimetic are shown in Table 18.

[0973] [Table 18-1]

[0974] [Table 18-2]

[0975] [Table 18-3]

[0976] [Table 19-1]

[0977] [Table 19-2]

[0978] General Procedure H for Amide Coupling to Obtain N-Fmoc-Tagged 5-nAA To a mixture of Tag5 or Tag5-(n-1)AA (1 eq.) in DCM (50 V) was added AAn (1.5 eq.), HOAt (1.5 eq.), and DIC (1.5 eq.). The resulting suspension was stirred at 20 °C for 2 h. A small sample of the reaction mixture was removed, suspended in ACN, and centrifuged twice (6000 rpm for 3 min). The precipitate that formed was dissolved in one drop of DCM, cleaved with TFA / TIPS / HO (95 / 2.5 / 2.5), diluted with MeOH / HO (2 / 1), and filtered. The filtrate was checked by LCMS to confirm the desired mass. The reaction was concentrated under reduced pressure to give a residue that was added to ACN / MeOH (2 / 1, 60 V) and stirred for 1 h. The suspension was filtered and washed twice with ACN (10 V). The precipitate was collected and used directly in the next step without further purification. The desired compound N-Fmoc-tagged 5-nAA was obtained as a white solid.

[0979] General Procedure I for Fmoc Deprotection to Prepare Tag 5-nAA A clear solution of N-Fmoc-tagged 5-nAA (1 eq.) in 20% piperidine / THF (20 V) was stirred at 50 °C for 0.5 h. A small sample was suspended in a CAN and centrifuged twice (6000 rpm for 3 min). The precipitate was collected, redissolved in one drop of DCM, cleaved with TFA / TIPS / HO (95 / 2.5 / 2.5), diluted with MeOH / HO (2 / 1), and filtered. The filtrate was checked by LCMS to confirm the completion of the reaction. The reaction was concentrated under reduced pressure to give a residue, which was added to ACN / MeOH (2 / 1, 60 V) and stirred for 1 h. The suspension was filtered and washed twice with ACN (10 V). The precipitate was collected and used directly in the next step without further purification. The desired compound, tagged 5-nAA, was obtained as a white solid.

[0980] Step a: Synthesis of 9H-fluoren-9-ylmethyl N-[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]carbamate (N-Fmoc-tag 5-1AA)

[0981] [ka] The reaction was carried out according to general procedure H for amide coupling using Tag 5 (1 g, 409.15 μmol, 1 eq), AA1 (272 mg, 0.62 mmol, 1.5 eq), HOAt (84 mg, 0.62 mmol, 1.5 eq), and DIC (77 mg, 0.62 mmol, 95.1 μL, 1.5 eq). The reaction was monitored by LC-MS. N-Fmoc-Tag 5-1AA (1.18 g, 344.59 μmol, 84.22% yield) was obtained as a white solid. LC-MS: RT = 5.17 min, C 28 H 30 Calculated mass of N2O3H: 443.24, measured m / z: 443.2 [M-tag + 2H] + .

[0982] Step b: Synthesis of (2S)-2-amino-N-[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]-5-(3,5-dimethylphenyl)pentanamide (TAG5-1AA)

[0983] [ka] The reaction was carried out according to General Procedure I for Fmoc deprotection. A clear solution of compound N-Fmoc-tag 5-1AA (1.16 g, 404.24 μmol, 1 eq) in 20% piperidine / THF (20 mL) was stirred at 50° C. for 0.5 hr. The reaction was monitored by LC-MS. Tag 5-1AA (1.06 g, 400.40 μmol, 99.05% yield) was obtained as a white solid. LC-MS: RT=2.10 min, C 13 H 20 Calculated mass of N2OH: 221.17, observed m / z: 221.1 [M-tag + 2H] + .

[0984] Step c: Synthesis of 9H-fluoren-9-ylmethyl N-[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]carbamate (N-Fmoc-tag 5-2AA)

[0985] [ka] The reaction was carried out according to general procedure H for amide coupling. To a mixture of compound tagged 5-1AA (1.06 g, 400.40 μmol, 1 eq) in DCM (40 mL) was added AA2 (363 mg, 600.6 μmol, 1.5 eq), HOAt (81.7 mg, 0.6 mmol, 84 μL, 1.5 eq), and DIC (76 mg, 0.6 mmol, 93 μL, 1.5 eq). The resulting suspension was stirred at 20 °C for 2 h. N-Fmoc-tag 5-2AA (1.25 g, 386.51 μmol, 96.53% yield) was obtained as a white solid. LC-(ESI):MS RT=6.72 min, C 49 H 54Calculated mass of N6O5H: 807.43, measured m / z: 807.4 [M-tag + 2H] + .

[0986] Step d: Synthesis of (2S)-2-[[(2S)-2-amino-3-[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]propanoyl]amino]-N-[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]-5-(3,5-dimethylphenyl)pentanamide (TAG5-2AA)

[0987] [ka] The reaction was carried out according to General Procedure I for Fmoc deprotection. A clear solution of compound N-Fmoc-tag 5-2AA (1.25 g, 386.51 μmol, 1 eq) in 20% piperidine / THF (25 mL) was stirred at 50° C. for 0.5 hr. Tag 5-2AA (1.15 g, crude) was obtained as a white solid. LC-MS: RT=4.46 min, C 34 H 44 Calculated mass of N6O3H: 585.36, measured m / z: 585.4 [M-tag + 2H] + .

[0988] Step e: Synthesis of tert-butyl (3S)-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-3-(9H-fluoren-9-ylmethoxycarbonylamino)-4-oxo-butanoate (N-Fmoc-tagged 5-3AA)

[0989] [ka] The reaction was carried out according to general procedure H for amide coupling. To a mixture of compound tagged 5-2AA (1.15 g, 381.83 μmol, 1 eq) in DCM (60 mL) was added AA3 (235.5 mg, 0.57 μmol, 1.5 eq), HOAt (78 mg, 0.57 mmol, 80.1 μL, 1.5 eq), and DIC (72.3 mg, 0.57 mmol, 88.7 μL, 1.5 eq). The resulting suspension was stirred at 20 °C for 2 h. N-Fmoc-tag 5-3AA (1.3 g, 381.76 μmol, 99.98% yield) was obtained as a white solid.

[0990] LCMS conditions (CR): Reversed-phase LCMS was performed using a NanoChrom ChromCore 120 C18 3µm column, 3.0 µm, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30 mm column.

[0991] LC-MS: RT=6.25 min, C 53 H 59 Calculated mass of N7O8H: 922.45, observed m / z: 922.4 [M-tag-tBu+3H] + .

[0992] LCMS conditions (BF): Reverse-phase LCMS was performed with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. Xtimate C18 2.1 * It was performed using a 30mm 3um column.

[0993] Step f: Synthesis of tert-butyl (3S)-3-amino-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG5-3AA)

[0994] [ka] The reaction was carried out according to General Procedure I for Fmoc deprotection. A clear solution of compound N-Fmoc-tag 5-3AA (1.30 g, 381.76 μmol, 1 eq) in 20% piperidine / THF (26 mL) was stirred at 50° C. for 0.5 hr. Compound tag 5-3AA (1.2 g, 377.00 μmol, 98.75% yield) was obtained as a white solid.

[0995] LC-MS: RT=4.646 min, C 38 H 49 Calculated mass of N7O6H: 700.38, observed m / z: 700.5 [M-tag-tBu+3H] + ;RT=5.065 minutes, C 42 H 57 Calculated mass of N7O6Na: 778.43, measured m / z: 778.5 [M-tag + 2H + Na] + .

[0996] Step g: tert-Butyl(3S)-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl Synthesis of ]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-3-[[(2S)-3-[tert-butyl(dimethyl)silyl]oxy-2-(9H-fluoren-9-ylmethoxycarbonylamino)propanoyl]amino]-4-oxo-butanoate (N-Fmoc-tag 5-4AA)

[0997] [ka] The reaction was carried out according to General Procedure H for amide coupling. To a mixture of compound N-Fmoc-tag 5-3AA (1.1 g, 345.59 μmol, 1 eq) in DCM (50 mL) was added TBS-AA4 (229 mg, 0.52 mmol, 1.5 eq), HOAt (70.5 mg, 0.52 mmol, 1.5 eq), and DIC (65.4 mg, 0.52 mmol, 80.3 μL, 1.5 eq). The resulting suspension was stirred at 20 °C for 2 h. The reaction was monitored by LC-MS. Compound N-Fmoc-tag 5-4AA (1.06 g, 293.91 μmol, 85.05% yield) was obtained as a white solid.

[0998] LC-MS: RT=6.01 min, C 56 H 64 N8O 10 Calculated mass of H: 1009.48, observed m / z: 1009.4 [M-tag-tBu-TBS+4H] + LCMS conditions (BF): Reverse-phase LCMS was performed using an Xtimate C18 2.1 column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. *It was performed using a 30mm 3um column.

[0999] Step h: Synthesis of tert-butyl (3S)-3-[[(2S)-2-amino-3-[tert-butyl(dimethyl)silyl]oxy-propanoyl]amino]-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG5-4AA)

[1000] [ka] A clear solution of compound N-Fmoc-tag 5-4AA (1.05 g, 291.13 μmol, 1 eq) in 20% piperidine / THF (20 mL) was stirred for 0.5 hr at 50° C. Following the general precipitation procedure, the desired compound tag 5-4AA (985 mg, 291.05 μmol, 99.97% yield) was obtained as a white solid.

[1001] LC-MS: RT=4.24 min, C 41 H 54 Calculated mass of N8O8H: 787.42, observed m / z: 787.4 [M-tag-tBu-TBS+4H] + ;RT=5.10 minutes, C 47 H 68 Calculated mass of N8O8SiH: 901.50, measured m / z: 901.5 [M-tag-tBu+3H] + LCMS conditions (BF): Reverse-phase LCMS was performed using an Xtimate C18 2.1 column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30mm 3um column.

[1002] Step i: tert-butyl(3S)-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carba Synthesis of N-Fmoc-tagged 5-5AA (N-Fmoc-tag 5-5AA)

[1003] [ka] To a mixture of compound N-Fmoc-tag 5-4AA (985 mg, 291.05 μmol, 1 eq) in DCM (50 mL), TBS-AA5 (2S,3R)-3-[tert-butyl(dimethyl)silyl]oxy-2-(9H-fluoren-9-ylmethoxycarbonylamino)butanoic acid (199 mg, 0.44 mmol, 1.5 eq), HOAt (59.4 mg, 0.44 mmol, 1.5 eq), and DIC (55 mg, 0.44 mmol, 67.6 μL, 1.5 eq) were added. The resulting suspension was stirred at 20 °C for 2 h. Following the general precipitation procedure, compound N-Fmoc-tag 5-5AA (1.05 g, 274.73 μmol, 94.39% yield) was obtained as a white solid.

[1004] LC-MS: RT=5.91 min, C 60 H 71 N9O 12 Calculated mass of H: 1110.53, observed m / z: 1110.6 [M-tag-tBu-2TBS+5H] +LCMS conditions (BF): Reverse-phase LCMS was performed using an Xtimate C18 2.1 column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30mm 3um column.

[1005] Step j: Synthesis of tert-butyl (3S)-3-[[(2S)-2-amino-3-[tert-butyl(dimethyl)silyl]oxy-propanoyl]amino]-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG5-5AA)

[1006] [ka] A clear solution of compound N-Fmoc-tag 5-5AA (1.05 g, 291.13 μmol, 1 eq) in 20% piperidine / THF (20 mL) was stirred at 50 °C for 0.5 hr. Following the general precipitation procedure, the desired compound tag 5-5AA (0.83 g, 245.25 μmol, 84.24% yield) was obtained as a white solid. LC-MS: RT = 5.107 min, C 51 H 75 N9O 10 Calculated mass of SiH: 1002.55, measured m / z: 1002.7 [M-tag-tBu-TBS+4H] + LCMS conditions (CR): Reversed-phase LCMS was performed using a NanoChrom ChromCore 120 C18 3µm column, 3.0 µm column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. *It was performed using a 30 mm column.

[1007] Step k: tert-butyl(3S)-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl) Synthesis of N-Fmoc-tag 5-6AA (butyl]amino)-2-oxo-ethyl]amino)-3-[[(2S)-3-[tert-butyl(dimethyl)silyl]oxy-2-[[(2S,3R)-3-[tert-butyl(dimethyl)silyl]oxy-2-[[(2S)-2-(9H-fluoren-9-ylmethoxycarbonylamino)-3-(2-fluorophenyl)-2-methyl-propanoyl]amino]butanoyl]amino]propanoyl]amino]-4-oxo-butanoate (N-Fmoc-tag 5-6AA)

[1008] [ka] To a mixture of compound N-Fmoc-tag 5-5AA (0.83 g, 245.25 μmol, 1 eq) in DCM (50 mL), AA6 (2S)-2-(9H-fluoren-9-ylmethoxycarbonylamino)-3-(2-fluorophenyl)-2-methyl-propanoic acid (154.3 mg, 367.9 μmol, 1.5 eq), HOAt (84 mg, 0.62 mmol, 1.5 eq), and DIC (78 mg, 0.62 mmol, 96 μL, 1.5 eq) were added. The resulting suspension was stirred at 20 °C for 2 h. Following the general precipitation procedure, the desired compound N-Fmoc-tag 5-6AA (0.95 g, crude material) was obtained as a white solid.

[1009] LC-MS: RT=6.46 min, C 70 H 81 FN 10 O 13Calculated mass of H: 1289.61, observed m / z: 1290.8 [M-tag-tBu-2TBS+5H] + LCMS conditions (BV): Reverse phase LCMS was performed using a NanoChrom ChromCore 120 C18 3µm column, 3.0 µm column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30 mm column.

[1010] LC-MS: RT=6.25 min, C 70 H 81 FN 10 O 13 Calculated mass of H: 1289.61, observed m / z: 1289.5 [M-tag-tBu-2TBS+5H] + LCMS conditions (BF): Reverse-phase LCMS was performed using an Xtimate C18 2.1 column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30mm 3um column.

[1011] Step 1: tert-Butyl (3S)-3-[[(2S)-2-[[(2S,3R)-2-[[(2S)-2-amino-3-(2-fluorophenyl)-2-methyl-propanoyl]amino]-3-[tert-butyl(dimethyl)silyl]oxy-butanoyl]amino]-3-[tert-butyl(dimethyl)silyl]oxy-propanoyl]amino]-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl Synthesis of]nyl]methyl]-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG5-6AA)

[1012] [ka] A clear solution of compound N-Fmoc-tag 5-6AA (0.95 g, 237.43 μmol, 1 eq) in 20% piperidine / THF (20 mL) was stirred for 0.5 hr at 50° C. Following the general precipitation procedure, compound tag 5-6AA (0.85 g, 224.93 μmol, 94.74% yield) was obtained as a white solid.

[1013] LC-MS: RT=4.64 min, C 55 H 71 FN 10 O 11 Calculated mass of H: 1067.54, observed m / z: 1067.7 [M-tag-tBu-2TBS+5H] + LCMS conditions (CR): Reversed-phase LCMS was performed using a NanoChrom ChromCore 120 C18 3µm column, 3.0 µm column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30 mm column.

[1014] Step M: tert-butyl(3S)-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carba Synthesis of N-Fmoc-tag 5-7AA (N-Fmoc-tag 5-7AA)

[1015] [ka] To a mixture of compound N-Tag 5-6AA (0.85 g, 224.93 μmol, 1 eq) in DCM (50 mL), TBS-AA7 (2S,3R)-3-[tert-butyl(dimethyl)silyl]oxy-2-(9H-fluoren-9-ylmethoxycarbonylamino)butanoic acid (1.02 g, 2.25 mmol, 10 eq), HOAt (306.16 mg, 2.25 mmol, 314.65 μL, 10 eq), and DIC (283.87 mg, 2.25 mmol, 348.30 μL, 10 eq) were added. The resulting suspension was stirred at 40 °C for 12 h. Following the general precipitation procedure, compound N-Fmoc-Tag 5-7AA (0.91 g, crude material) was obtained as a white solid.

[1016] LC-MS: RT=6.24 min, C 74 H 88 FN 11 O 15 Calculated mass of H: 1390.65, observed m / z: 1390.5 [M-tag-tBu-3TBS+6H] + LCMS conditions (BF): Reverse-phase LCMS was performed using an Xtimate C18 2.1 column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30mm 3um column.

[1017] Step n: tert-Butyl (3S)-3-[[(2S)-2-[[(2S,3R)-2-amino-3-[tert-butyl(dimethyl)silyl]oxy-butanoyl]amino]-3-[tert-butyl(dimethyl)silyl]oxy-propanoyl]amino]-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4- Synthesis of [2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate;H2O (Tag 5-7AA)

[1018] [ka] A clear solution of compound N-Fmoc-tag 5-7AA (0.91 g, 238.10 μmol, 1 eq) in 20% piperidine / THF (3 mL) was stirred for 0.5 hr at 50° C. Following the general precipitation procedure, compound tag 5-7AA (0.85 g, 236.13 μmol, 99.17% yield) was prepared as a white solid.

[1019] LC-MS: RT=5.72 min, C 65 H 92 FN 11 O 13 Calculated mass of SiH: 1282.67, observed m / z: 1282.7 [M-tag-tBu-2TBS+5H] + LCMS conditions (CR): Reversed-phase LCMS was performed using a NanoChrom ChromCore 120 C18 3µm column, 3.0 µm column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30 mm column.

[1020] Step o: tert-Butyl(3S)-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-3-[ Synthesis of [(2S)-3-[tert-butyl(dimethyl)silyl]oxy-2-[[(2S,3R)-3-[tert-butyl(dimethyl)silyl]oxy-2-[[2-[[(2S,3R)-3-[tert-butyl(dimethyl)silyl]oxy-2-[[2-(9H-fluoren-9-ylmethoxycarbonylamino)acetyl]amino]butanoyl]amino]-3-(2-fluorophenyl)-2-methyl-propanoyl]amino]butanoyl]amino]propanoyl]amino]-4-oxo-butanoate (N-Fmoc-tag 5-8AA)

[1021] [ka] To a mixture of compound N-Fmoc-tag 5-7AA (0.85 g, 236.13 μmol, 1 eq) in DCM (50 mL), 2-(9H-fluoren-9-ylmethoxycarbonylamino)acetic acid (AA8) (105.3 mg, 354 μmol, 1.5 eq), HOAt (48.2 mg, 354 μmol, 1.5 eq), and DIC (44.7 mg, 354 μmol, 54.8 μL, 1.5 eq) were added. The resulting suspension was stirred at 20 °C for 2 h. Following the general precipitation procedure, the desired compound N-Fmoc-tag 5-8AA (0.79 g, 184.86 μmol, 78.29% yield) was obtained as a white solid.

[1022] LC-MS: RT=6.32 min, C 76 H 91 FN 12 O 16Calculated mass of H: 1447.68, observed m / z: 1448.6 [M-tag-tBu-3TBS+6H] + LCMS conditions (BV): Reverse phase LCMS was performed using a NanoChrom ChromCore 120 C18 3µm column, 3.0 µm column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30 mm column.

[1023] LC-MS: RT=6.11 min, C 76 H 91 FN 12 O 16 Calculated mass of H: 1447.68, observed m / z: 1447.6 [M-tag-tBu-3TBS+6H] + LCMS conditions (BF): Reverse-phase LCMS was performed using an Xtimate C18 2.1 column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30mm 3um column.

[1024] Step p: tert-Butyl(3S)-3-[[(2S)-2-[[(2S,3R)-2-[[2-[[(2S,3R)-2-[(2-aminoacetyl)amino]-3-[tert-butyl(dimethyl)silyl]oxy-butanoyl]amino]-3-(2-fluorophenyl)-2-methyl-propanoyl]amino]-3-[tert-butyl(dimethyl)silyl]oxy-butanoyl]amino]-3-[tert-butyl(dimethyl)silyl]oxy-propanoyl]amino]-4-[[(1S)-1-[[4- Synthesis of [4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG5-8AA)

[1025] [ka] A clear solution of compound N-Fmoc-tag 5-8AA (0.79 g, 184.86 μmol, 1 eq) in 20% piperidine / THF (20 mL) was stirred at 50 °C for 0.5 hr. Following the general precipitation procedure, compound tag 5-8AA (0.74 g, 182.66 μmol, 98.81% yield) was obtained as a white solid. LC-MS: RT = 4.47 min, C 61 H 81 FN 12 O 14 Calculated mass of H: 1225.61, observed m / z: 1225.6 [M-tag-tBu-3TBS+6H] + .

[1026] Step Q: tert-butyl(3S)-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-3-[[(2S)-3-[tert-butyl(dimethyl Synthesis of N-Fmoc-tagged 5-9AA) of N-(2-(9H-fluoren-9-ylmethoxycarbonylamino)-3-(1H-triazol-5-yl)propanoyl)amino]acetyl]amino]butanoyl]amino]-4-oxo-butanoate (N-Fmoc-tagged 5-9AA)

[1027] [ka] Use of protected AA9. To a mixture of compound Tag 5-8AA (50 mg, 12.34 μmol, 1 eq) in DCM (5 mL) was added AA9 (7 mg, 18.51 μmol, 1.5 eq), HOAt (2.52 mg, 18.51 μmol, 1.5 eq), and DIC (2.34 mg, 18.51 μmol, 2.87 μL, 1.5 eq). The resulting suspension was stirred at 20 °C for 2 h. The desired mass was not detected (Table 19).

[1028] [Table 20]

[1029] LCMS conditions (CR): Reversed-phase LCMS was performed using a NanoChrom ChromCore 120 C18 3µm column, 3.0 µm, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30 mm column.

[1030] LCMS conditions (BF): Reverse-phase LCMS was performed with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. Xtimate C18 2.1 * It was performed using a 30mm 3um column.

[1031] Use of protected DMB-AA9. To a mixture of compound tagged 5-8AA (0.5 g, 123.42 μmol, 1 eq) in DCM (5 mL), DMB-AA9 (98 mg, 185 μmol, 1.5 eq), HOAt (25.2 mg, 185 μmol, 1.5 eq), and DIC (23.36 mg, 185 μmol, 28.6 μL, 1.5 eq) were added. The resulting suspension was stirred at 20 °C for 2 h. Following the general precipitation procedure, the desired compound N-Fmoc-tag 5-9AA (510 mg, 111.77 μmol, 90.57% yield) was obtained as a white solid (Table 19).

[1032] LC-MS: RT=5.91 min, C 80 H 96 FN 17 O 17 Calculated mass of H: 1586.74 [M-tag-tBu-3TBS-DMB+7H] + , C 80 H 96 FN 17 O 17 Calculated mass of H2: 793.87 [M-tag-tBu-3TBS-DMB+8H] 2+ , m / z measured value 794.3 [M-tag-tBu-3TBS-DMB+8H] 2+LCMS conditions (BV): Reverse phase LCMS was performed using a NanoChrom ChromCore 120 C18 3µm column, 3.0 µm column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30 mm column.

[1033] LC-MS: RT=5.68 min, C 80 H 96 FN 17 O 17 Calculated mass of H: 1586.74 [M-tag-tBu-3TBS-DMB+7H] + , C 80 H 96 FN 17 O 17 Calculated mass of H2: 793.87 [M-tag-tBu-3TBS-DMB+8H] 2+ , m / z measured value 794.2 [M-tag-tBu-3TBS-DMB+8H] 2+ LCMS conditions (BF): Reverse-phase LCMS was performed using an Xtimate C18 2.1 column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30mm 3um column.

[1034] Step R: tert-butyl(3S)-3-[[(2S)-2-[[(2S,3R)-2-[[2-[[(2S,3R)-2-[[2-[[(2S)-2-amino-3-[1-[(2,4-dimethoxyphenyl)methyl]tetrazol-5-yl]propanoyl]amino]acetyl]amino]-3-[tert-butyl(dimethyl)silyl]oxy-butanoyl]amino]-3-(2-fluorophenyl)-2-methyl-propanoyl]amino]-3-[tert-butyl(dimethyl)silyl]oxy-butanoyl]amino]-3-[tert-butyl(dimethyl)silyl]oxy Synthesis of [[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-4-oxo-butanoate (TAG5-9AA)

[1035] [ka] A clear solution of compound N-Fmoc-tag 5-9AA (500 mg, 109.58 μmol, 1 eq.) in 20% piperidine / THF (20 mL) was stirred for 0.5 hr at 50° C. Following the general precipitation procedure, the desired compound tag 5-9AA (0.47 g, 108.28 μmol, 98.81% yield) was obtained as a white solid.

[1036] LC-MS: RT=4.78 min, C 65 H 86 FN 17 O 15 Calculated mass of H: 1364.66, observed m / z: 1364.7 [M-tag-tBu-3TBS-DMB+7H] +LCMS conditions (CR): Reversed-phase LCMS was performed using a NanoChrom ChromCore 120 C18 3µm column, 3.0 µm column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30 mm column.

[1037] LC-MS: RT=4.47 min, C 65 H 86 FN 17 O 15 Calculated mass of H: 1364.66, observed m / z: 1364.7 [M-tag-tBu-3TBS-DMB+7H] + LCMS conditions (BF): Reverse-phase LCMS was performed using an Xtimate C18 2.1 column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30mm 3um column.

[1038] Step S: tert-butyl(3S)-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-3-[[(2S)-3-[tert-butyl(dimethyl)silyl]oxy-2-[[(2S,3R) Synthesis of -3-[tert-butyl(dimethyl)silyl]oxy-2-[[2-[[(2S,3R)-3-[tert-butyl(dimethyl)silyl]oxy-2-[[2-[[(2S)-3-[1-[(2,4-dimethoxyphenyl)methyl]tetrazol-5-yl]-2-[[2,2-dimethyl-3-oxo-3-[2-(3-tritylimidazol-4-yl)ethylamino]propanoyl]amino]propanoyl]amino]acetyl]amino]butanoyl]amino]-3-(2-fluorophenyl)-2-methyl-propanoyl]amino]butanoyl]amino]propanoyl]amino]-4-oxo-butanoate (TAG 5-10AA)

[1039] [ka] To a mixture of compound Tag 5-9AA (450 mg, 103.67 μmol, 1 eq) in DCM (20 mL) was added AA10 (72.73 mg, 155.5 μmol, 1.5 eq), HOAt (21.2 mg, 155.5 μmol, 1.5 eq), and DIC (19.6 mg, 155.5 μmol, 24.1 μL, 1.5 eq). The resulting suspension was stirred at 20 °C for 2 h. Following the general precipitation procedure, the desired compound Tag 5-10AA (480 mg, 100.21 μmol, 96.66% yield) was obtained as a white solid. LC-MS (Method 4): RT = 4.456 min, C 75 H 99 FN 20 O 17Calculated mass of H: 1571.76 [M-tag-tBu-3TBS-DMB-Trt+8H] + , C 75 H 99 FN 20 O 17 Calculated mass of H2: 786.38 [M-tag-tBu-3TBS-DMB-Trt+9H] 2+ , m / z measured value 786.7 [M-tag-tBu-3TBS-DMB-Trt+9H] 2+ ;LC-MS (Method 4): RT=4.85 min, C 75 H 99 FN 20 O 17 Calculated mass of H: 1571.76 [M-tag-tBu-3TBS-DMB-Trt+8H] + , C 75 H 99 FN 20 O 17 Calculated mass of H2: 786.38 [M-tag-tBu-3TBS-DMB-Trt+9H] 2+ , m / z measured value 786.8 [M-tag-tBu-3TBS-DMB-Trt+9H] 2+ .

[1040] Step T: tert-Butyl(3S)-4-[[(1S)-2-[[(1S)-1-[[[4-[2-[bis[[3,4,5-tris(docosa-1,3,5,7,9,11,13,15,17,19,21-undecainoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamoyl]-4-(3,5- Dimethylphenyl)butyl]amino]-1-[[4-[4-[4-[4-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(tert-butoxycarbonylamino)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxymethyl]triazol-1-yl]butoxy]-2-ethyl-phenyl]phenyl]methyl]-2-oxo-ethyl] Synthesis of amino]-3-[[(2S)-3-[tert-butyl(dimethyl)silyl]oxy-2-[[(2S,3R)-3-[tert-butyl(dimethyl)silyl]oxy-2-[[2-[[(2S,3R)-3-[tert-butyl(dimethyl)silyl]oxy-2-[[2-[[(2S)-3-[1-[(2,4-dimethoxyphenyl)methyl]tetrazol-5-yl]-2-[[2,2-dimethyl-3-oxo-3-[2-(3-tritylimidazol-4-yl)ethylamino]propanoyl]amino]propanoyl]amino]acetyl]amino]butanoyl]amino]-3-(2-fluorophenyl)-2-methyl-propanoyl]amino]butanoyl]amino]propanoyl]amino]-4-oxo-butanoate (Tag5-M1)

[1041] [ka] Compound Tag 5-10AA (0.3 g, 62.63 μmol, 1 eq) was dissolved in HO (1 mL) and THF (4 mL). Then, a solution of CuSO4.5HO (23.46 mg, 93.94 μmol, 1.5 eq) and sodium ascorbate (18.61 mg, 93.94 μmol, 1.5 eq) in HO (1 mL), a solution of TBTA (16.62 mg, 31.31 μmol, 0.5 eq) in DMSO (0.5 mL), and a solution of tert-butyl PEG-linker (95.37 mg, 187.89 μmol, 3 eq) in THF (4 mL) were added. The mixture was stirred at 40 °C for 12 h. Following the general precipitation procedure, the desired compound Tag 5-M1 (330 mg, 62.29 μmol, 99.46% yield) was obtained as a white solid.

[1042] LC-MS: RT=3.72 min, C 94 H 136 FN 21 O 25 Calculated mass of H2: 990.01 [M-tag-tBu-3TBS-DMB-Trt-Boc + 10H] 2+ , m / z measured value 990.6 [M-tag-tBu-3TBS-DMB-Trt-Boc+10H] 2+ LCMS conditions (CR): Reversed-phase LCMS was performed using a NanoChrom ChromCore 120 C18 3µm column, 3.0 µm column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30 mm column.

[1043] LC-MS: RT=3.39 min, C 94 H 136 FN 21 O 25 Calculated mass of H2: 990.01, observed m / z: 990.4 [M-tag-tBu-3TBS-DMB-Trt-Boc+10H] 2+ ;RT=3.99 minutes, C 113 H 150 FN 21 O 25Calculated mass of H2: 1111.06, observed m / z: 1111.5 [M-tag-tBu-3TBS-DMB-Boc+9H] 2+ LCMS conditions (BF): Reverse-phase LCMS was performed using an Xtimate C18 2.1 column, eluting with a 10% to 80% gradient of ACN containing 0.02% TFA (solvent B) and water containing 0.04% TFA (solvent A) at a flow rate of 1.5 mL / min. * It was performed using a 30mm 3um column.

[1044] Step u: (3S)-4-[[(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[(1S)-1-carbamoyl-4-(3,5-dimethylphenyl)butyl]amino]-2-oxo-ethyl]amino]-3-[[(2S)-2-[[(2S,3R)-2-[[3-(2-fluorophenyl)-2-[[(2S,3R)-3-hydroxy-2-[[2-[[ Synthesis of (2S)-2-[[3-[2-(1H-imidazol-5-yl)ethylamino]-2,2-dimethyl-3-oxo-propanoyl]amino]-3-(1H-tetrazol-5-yl)propanoyl]amino]acetyl]amino]butanoyl]amino]-2-methyl-propanoyl]amino]-3-hydroxy-butanoyl]amino]-3-hydroxy-propanoyl]amino]-4-oxo-butanoic acid (M1)

[1045] [ka] To a mixture of tag 5-M1 (430 mg, 81.17 μmol, 1 eq) bearing all side chain protecting groups in DCM (10 mL) was added TFA cocktail (TFA / TIPS / HO=95 / 2.5 / 2.5) (30 mL). The resulting suspension was stirred at 20 °C for 2 hr. A sample was removed, suspended in PE / MTBE (3 / 1, 1 mL), and centrifuged to give a solid. The solid was diluted with MeOH and HO, filtered, and the filtrate was examined by LCMS, which showed the desired mass of M1. The reaction was concentrated under reduced pressure to give a residue, which was added to PE / TBME (3 / 1, 20 V) and stirred for 1 h. The suspension was filtered to give a white solid. The residue was purified by prep-HPLC (basic conditions): Column: Boston Prime C18 150 * 30mm * Purification was performed by HPLC using a 5 μm column; mobile phase: [water (ammonia hydroxide v / v)-ACN]; gradient: 25% to 45% B over 11 min. The desired compound M1 (5.1 mg, 2.45 μmol, 3.02% yield, 95% purity) was obtained as a white solid. LC-MS: RT = 3.399 min, C 94 H 136 N 21 O 25 Calculated mass of FH: 1979.01 [M+H] + , C 94 H 136 N 21 O 25 Calculated mass of FH2: 990.01 [M+2H] 2+ , m / z measured value 990.5 [M+2H] 2+ ;LCMS conditions: Method 7.

[1046] M1, 1 H NMR, 19 FNMR (non-TFA salt) and M1 (TFA salt), F NMR, 13M1 was characterized using C NMR as well as UPLC. UPLC of M1 showed an impurity at RT = 6.5 min, which was a system-related impurity. LCMS of M1 showed that the major peak at retention time 1.564 min corresponded to the desired product M1, with observed masses of [M+H] + = 1980.3, [M / 2+H] + = 990.3, [M / 3+H] + = 660.7, [M / 4+H] + = 495.8.

[1047] Confirmation of M1-HCl salt by IC showed that the high quality content of HCl was 2.08% and the molar content was 1.15 moleq.

[1048] 8.4 Synthesis of M3 into Tag 5 (LPPS) The synthesis of M3 to tag5 is shown here. Figure 7 shows the synthesis of tag5-M3, and Figure 12 shows the extension conditions and results for making M3 using tag5.

[1049] Step a: Synthesis of 9H-fluoren-9-ylmethyl N-[rac-(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[[4-[2-[bis[4-[[3,4,5-tri(docosoxy)phenyl]methoxy]phenyl]methylamino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]amino]-2-oxo-ethyl]carbamate (N-Fmoc-tag 5-2AA)

[1050] [ka] To a mixture of compound tag 5 (2 g, 757 μmol, 1 eq.) in DCM (50 mL) was added AA2 (687 mg, 1.14 mmol, 1.5 eq.) in THF (40 mL), HOAt (155 mg, 1.14 mmol, 1.5 eq.), and DIC (143.28 mg, 1.14 mmol, 175.81 μL, 1.5 eq.). The solution was then stirred at 50 °C for 12 h and monitored by LC-MS trace. The reaction was carried out using General Procedure H for amide coupling to give N-Fmoc-tag 5-2AA (2.3 g, 712.30 μmol, 94.11% yield) as a white solid. LC-MS: RT = 5.506 min, C 36 H 37 Calculated mass of N5O4Na: 626.03, measured m / z: 626.70 [M-tag + H + Na] + .

[1051] Step b: Synthesis of rac-(2S)-2-amino-3-[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]-N,N-bis[[3,4,5-tri(docosoxy)phenyl]methyl]propanamide (TAG5-2AA)

[1052] [ka] A clear solution of compound N-Fmoc-tag 5-2AA (1 g, 366.11 μmol, 1 eq) in 20% piperidine / THF (20 mL) was stirred at 50° C. for 2 h without monitoring. Following the general precipitation procedure, tag 5-2AA (0.9 g, 358.68 μmol, 97.97% yield) was obtained as a white solid.

[1053] Step c: Synthesis of tert-butyl rac-(3S)-3-(9H-fluoren-9-ylmethoxycarbonylamino)-4-oxo-4-[[rac-(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[[[4-[2-[bis[[3,4,5-tri(docosoxy)phenyl]methyl]amino]-2-oxo-ethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]amino]-2-oxo-ethyl]amino]butanoate (N-Fmoc-tagged 5-3AA)

[1054] [ka] To a mixture of compound N-Fmoc-tag 5-2AA (0.9 g, 320.45 μmol, 1 eq) in DCM (40 mL) was added AA3 (395.55 mg, 961.36 μmol, 3.0 eq), HOAt (130.85 mg, 961.36 μmol, 134.48 μL, 3.0 eq), and DIC (121.32 mg, 961.36 μmol, 148.86 μL, 3.0 eq). The resulting suspension was stirred at 20 °C for 2 h. Following the general precipitation procedure, the desired compound N-Fmoc-tag 5-3AA (1.2 g, crude material) was obtained as a white solid. LC-MS: RT = 5.038 min, C 40 H 43 Calculated mass of N6O7: 719.31, observed m / z: 719.30 [M-tag-tBu+2H] + .

[1055] Step d: Synthesis of tert-butyl rac-(3S)-3-amino-4-oxo-4-[[rac-(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[bis[[3,4,5-tri(docosoxy)phenyl]methyl]amino]-2-oxo-ethyl]amino]butanoate (TAG5-3AA)

[1056] [ka] A clear solution of compound N-Fmoc-tag 5-3AA (1.2 g, crude material) in 20% piperidine / THF (25 mL) was stirred at 50° C. for 2 hours without monitoring. Following the general precipitation procedure, the desired compound tag 5-3AA (1.11 g, 414.12 μmol, 100.00% yield) was obtained as a white solid.

[1057] Step e: Synthesis of tert-butyl rac-(3S)-4-oxo-4-[[rac-(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[bis[[3,4,5-tri(docosoxy)phenyl]methyl]amino]-2-oxo-ethyl]amino]-3-[[rac-(2S)-3-[tert-butyl(dimethyl)silyl]oxy-2-(9H-fluoren-9-ylmethoxycarbonylamino)propanoyl]amino]butanoate (N-Fmoc-tag 5-4AA)

[1058] [ka] To a mixture of compound N-Fmoc-tag 5-3AA (1.11 g, 414.12 μmol, 1 eq.) in DCM (60 mL), TBS-AA4 (548.61 mg, 1.24 mmol, 3.0 eq.), DIC (156.78 mg, 1.24 mmol, 192.37 μL, 3.0 eq.), and HOAt (169.10 mg, 1.24 mmol, 173.79 μL, 3.0 eq.) were added. Following the general precipitation procedure, the desired compound N-Fmoc-tag 5-4AA (1.2 g, 386.60 μmol, 93.36% yield) was obtained as a white solid. LC-MS: RT = 4.791 min, C 43 H 47 Calculated mass of N7O9Na2: 851.34, observed m / z: 851.70 [M-tag-tBu-OTBS+4H] + .

[1059] Step f: Synthesis of tert-butyl rac-(3S)-4-oxo-3-[[rac-(2S)-2-amino-3-[tert-butyl(dimethyl)silyl]oxy-propanoyl]amino]-4-[[rac-(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[bis[[3,4,5-tri(docosoxy)phenyl]methyl]amino]-2-oxo-ethyl]amino]butanoate (TAG5-4AA)

[1060] [ka] A clear solution of compound N-Fmoc-tag 5-4AA (1.2 g, crude material) in 20% piperidine / THF (25 mL) was stirred without monitoring at 50° C. for 2 h. Following the general precipitation procedure, the desired compound tag 5-4AA (1.1 g, 381.71 μmol, 98.74% yield) was obtained as a white solid.

[1061] Step g: Synthesis of tert-butyl rac-(3S)-4-oxo-4-[[rac-(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[bis[[3,4,5-tri(docosoxy)phenyl]methyl]amino]-2-oxo-ethyl]amino]-3-[[rac-(2S)-3-[tert-butyl(dimethyl)silyl]oxy-2-[[rac-(2S,3R)-3-[tert-butyl(dimethyl)silyl]oxy-2-(9H-fluoren-9-ylmethoxycarbonylamino)butanoyl]amino]propanoyl]amino]butanoate (N-Fmoc-tag 5-5AA)

[1062] [ka] To a mixture of compound N-Fmoc-tag 5-4AA (1.1 g, 381.71 μmol, 1 eq.) in THF (40 mL), TBS-AA5 (521.75 mg, 1.15 mmol, 3.0 eq.), DIC (144.52 mg, 1.15 mmol, 177.32 μL, 3.0 eq.), and HOAt (155.87 mg, 1.15 mmol, 160.19 μL, 3.0 eq.) were added. The resulting suspension was stirred at 20 °C for 2 h and monitored by LC-MS. Following the general precipitation procedure, the desired compound N-Fmoc-tag 5-5AA (1.25 g, 376.58 μmol, 98.66% yield) was obtained as a white solid. LC-MS: RT = 4.670 min, C 47 H 55 N8O 11 Calculated mass of H: 907.39, observed m / z: 907.50 [M-tag-tBu-2OTBS+5H] + .

[1063] Step h: Synthesis of tert-butyl rac-(3S)-4-oxo-4-[[rac-(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[bis[[3,4,5-tri(docosoxy)phenyl]methyl]amino]-2-oxo-ethyl]amino]-3-[[rac-(2S)-3-[tert-butyl(dimethyl)silyl]oxy-2-[[rac-(2S,3R)-2-amino-3-[tert-butyl(dimethyl)silyl]oxy-butanoyl]amino]propanoyl]amino]butanoate (TAG5-5AA)

[1064] [ka] A clear solution of compound N-Fmoc-tag 5-5AA (1.25 g, 376.58 μmol, 1 eq.) in 20% piperidine / THF (20 mL) was stirred for 2 h at 50° C. Following the general precipitation procedure, the desired compound tag 5-5AA (1.1 g, crude material) was obtained as a white solid.

[1065] Step i: Synthesis of tert-butyl rac-(3S)-4-oxo-4-[[rac-(1S)-1-[[4-[4-(4-azidobutoxy)-2-ethyl-phenyl]phenyl]methyl]-2-[bis[[3,4,5-tri(docosoxy)phenyl]methyl]amino]-2-oxo-ethyl]amino]-3-[[rac-(2S)-3-[tert-butyl(dimethyl)silyl]oxy-2-[[rac-(2S,3R)-3-[tert-butyl(dimethyl)silyl]oxy-2-[[2-(9H-fluoren-9-ylmethoxycarbonylamino)-3-(2-fluorophenyl)-2-methyl-propanoyl]amino]butanoyl]amino]propanoyl]amino]butanoate (N-Fmoc-tag 5-6AA)

[1066] [ka] To a mixture of compound N-Fmoc-tag 5-5AA (1.1 g, 355.17 μmol, 1 eq.) in THF (50 mL), AA6 (rac-(2S)-2-(9H-fluoren-9-ylmethoxycarbonylamino)-3-...

Claims

1. Formula (I) 【Chemistry 1】 [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R １ is C １～５ is alkyl, R ２ represents, independently at each occurrence, an optionally substituted C １０～３０ is alkyl, R ３ represents, independently at each occurrence, an optionally substituted C １０～３０ alkyl] A compound having the structure: or a salt thereof.

2. R １ The compound of claim 1 , wherein is methyl.

3. R ２ is, independently at each occurrence, optionally replaced by C １８～２６ alkyl, and R ３ is, independently at each occurrence, optionally replaced by C １８～２６ The compound of claim 1 , wherein the aryl group is alkyl.

4. Each R ２ are independently optionally substituted C ２２ alkyl, and each R ３ are independently optionally substituted C ２２ The compound of claim 1 , wherein the aryl group is alkyl.

5. The compound is represented by formula (Ia) 【Chemistry 2】 2. The compound of claim 1 having the structure:

6. Formula (I) 【Transformation 3】 [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R １ is C １～５ is alkyl, R ２ represents, independently at each occurrence, an optionally substituted C １０～３０ is alkyl, R ３ represents, independently at each occurrence, an optionally substituted C １０～３０ alkyl] or a salt thereof, comprising: (a) Structure 【Chemistry 4】 [In the formula, wherein PG is a suitable protecting group; and (b) forming a compound of formula (I) from a compound of formula (II); A process involving:

7. 7. The process of claim 6, wherein PG is selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

8. The compound of formula (II) is represented by the formula (IIa) 【Transformation 5】 7. The process of claim 6, having the structure:

9. wherein said step (b) of forming a compound of formula (I) comprises: reacting the compound of formula (II) with a deprotecting agent to produce a compound of formula (I); 7. The process of claim 6, comprising:

10. The deprotecting agent is Pd(PPh) ３ , PhSiH ３ , H ２ 10. The process of claim 9, wherein the amine is selected from the group consisting of methyl methyl ether, piperidine, and trifluoroacetic acid (TFA).

11. structure 【Transformation 6】 [In the formula, PG １ is a suitable protecting group, PG ２ is a suitable protecting group; and Before step (a), a step of forming a compound of formula (II) from a compound of formula (III) 7. The process of claim 6, further comprising:

12. PG １ is independently selected from the group consisting of tert-butyldimethylsilyl (TBS), trimethylsilyl (TMS), triethylsilyl (TES), tert-butyldiphenylsilyl (TBDPS), and triisopropylsilyl (TIPS).

13. PG ２ is independently selected from the group consisting of tert-butyldimethylsilyl (TBS), trimethylsilyl (TMS), triethylsilyl (TES), tert-butyldiphenylsilyl (TBDPS), and triisopropylsilyl (TIPS).

14. Each PG １ and each PG ２ The process of claim 11 , wherein

15. PG １ and P.G. ２ 12. The process of claim 11, wherein:

16. The compound of formula (III) is of formula (IIIa) 【Transformation 7】 12. The process of claim 11 having the structure:

17. said step of forming a compound of formula (II) prior to step (a) reacting the compound of formula (III) with a deprotecting agent to obtain a deprotected intermediate; The intermediate is reacted with a compound of formula (IV) R-LG(IV) [In the formula, R is an optionally substituted C １０～３０ is alkyl, and LG is a suitable leaving group, Producing a compound of formula (II) 12. The process of claim 11, comprising:

18. 18. The process of claim 17, wherein LG is selected from the group consisting of halogen, OTf, OMs, and OTs.

19. The deprotecting agent is tetrabutylammonium fluoride (TBAF), NH ４ F, SiF ４ , AcOH, HCl, LiAlH ４ , and K ２ CO ３ 18. The process of claim 17, selected from the group consisting of:

20. structure 【Transformation 8】 providing a compound of formula (V) having the formula: Forming a compound of formula (III) from a compound of formula (V) 12. The process of claim 11, further comprising:

21. said step of forming a compound of formula (III) The compound of formula (V) is reacted with a compound of formula (VIa) PG-X(VIa) or Formula (VIb) PG ２ O(VIb) [In the formula, X is Cl or Br, Producing a compound of formula (III) 21. The process of claim 20, comprising:

22. The compound of formula (VIa) or formula (VIb) is AllocCl, Alloc ２ O, Cbz ２ O, CbzCl, FmocCl, and Boc ２ 22. The process of claim 21 , wherein the compound is selected from the group consisting of: O.

23. structure 【Chemistry 9】 [In the formula, PG ３ is a suitable protecting group; and forming a compound of formula (V) from a compound of formula (VII) 21. The process of claim 20, further comprising:

24. PG ３ is selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

25. The compound of formula (VII) has the following structure: 【Chemistry 10】 24. The process of claim 23, comprising:

26. said step of forming a compound of formula (V) reacting the compound of formula (VII) with a deprotecting agent to produce a compound of formula (V).

24. The process of claim 23, comprising:

27. The deprotecting agent is Pd(PPh) ３ , PhSiH ３ , H ２ 27. The process of claim 26, wherein the amine is selected from the group consisting of piperidine and trifluoroacetic acid (TFA).

28. structure 【Chemistry 11】 providing a compound of formula (VIII) having the formula forming a compound of formula (VII) from a compound of formula (VIII) 24. The process of claim 23, further comprising:

29. said step of forming a compound of formula (VII) The compound of formula (VIII) is 【Chemistry 12】 with a compound of formula (IX) having the formula (VII) 29. The process of claim 28, comprising:

30. The compound of formula (VIII) has the following structure: 【Chemistry 13】 29. The process of claim 28, comprising:

31. The compound of formula (IX) has the following structure: 【Chemistry 14】 30. The process of claim 29, comprising:

32. structure 【Chemistry 15】 [In the formula, PG ４ is a suitable protecting group; and forming a compound of formula (VIII) from a compound of formula (X) 29. The process of claim 28, further comprising:

33. PG ４ is selected from the group consisting of allyl, allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

34. The compound of formula (X) has the following structure: 【Chemistry 16】 33. The process of claim 32, comprising:

35. said step of forming a compound of formula (VIII) reacting a compound of formula (X) with a deprotecting agent 33. The process of claim 32, comprising:

36. The deprotecting agent is Pd / K ２ CO ３ , 3-dimethylbarbituric acid / Pd(PPh ３ ) ４ , Pd(PPh) ３ , PhSiH ３ , H ２ 36. The process of claim 35, wherein the amine is selected from the group consisting of piperidine and trifluoroacetic acid (TFA).

37. structure 【Chemistry 17】 providing a compound of formula (XI) having the formula forming a compound of formula (X) from a compound of formula (XI) 33. The process of claim 32, further comprising:

38. The compound of formula (XI) has the following structure: [Chemistry 18] 38. The process of claim 37, comprising:

39. said step of forming a compound of formula (X) The compound of formula (XI) is reacted with a compound of formula (XII) 【Chemistry 19】 or a salt thereof, Producing a compound of formula (X) 38. The process of claim 37, comprising:

40. structure 【Chemistry 20】 providing a compound of formula (XIII) having the formula forming a compound of formula (XI) from a compound of formula (XIII) 38. The process of claim 37, further comprising:

41. The compound of formula (XIII) has the following structure: 【Chemistry 21】 41. The process of claim 40, comprising:

42. said step of forming a compound of formula (XI) reacting a compound of formula (XIII) with an oxidizing agent to produce a compound of formula (XI) 41. The process of claim 40, comprising:

43. The oxidizing agent is pyridinium chlorochromate (PCC), pyridinium dichromate (PDC), silver carbonate (Ag ２ CO ３ ), tetra-n-propylammonium perruthenate (TPAP), and Dess-Martin periodinane (DMP).

44. structure 【Chemistry 22】 [In the formula, Y is -OC １～６ wherein R is an alkyl group; and forming a compound of formula (XIII) from a compound of formula (XIV) 41. The process of claim 40, further comprising:

45. The compound of formula (XIV) has the following structure: 【Chemistry 23】 45. The process of claim 44, having the structure:

46. said step of forming a compound of formula (XIII) reacting a compound of formula (XIV) with a reducing agent 45. The process of claim 44, comprising:

47. The reducing agent is LiAlH ４ , DIBALH, and LiBH ４ 47. The process of claim 46, selected from the group consisting of:

48. structure 【Chemistry 24】 providing a compound of formula (XV) having the formula forming a compound of formula (XIV) from a compound of formula (XV) 45. The process of claim 44, further comprising:

49. The compound of formula (XV) has the following structure: 【Chemistry 25】 49. The process of claim 48, comprising:

50. said step of forming a compound of formula (XIV) reacting a compound of formula (XV) with a protecting group introducing agent to produce a compound of formula (XIV).

49. The process of claim 48, comprising:

51. 51. The process of claim 50, wherein the protecting group introducing agent is selected from the group consisting of TBSCl, TBSOTf, TMSCl, TMSOTf, TESCl, TESOTf, TBDPSCl, TBDPSOTf, TIPSCl, and TIPSOTf.

52. 52. A product prepared by the method of any one of claims 6 to 51.

53. The product is a compound of formula (I) 【Chemistry 26】 [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R １ is C １～５ is alkyl, R ２ represents, independently at each occurrence, an optionally substituted C １０～３０ is alkyl, R ３ represents, independently at each occurrence, an optionally substituted C １０～３０ alkyl] or a salt thereof.

54. The product is represented by formula (Ia) 【Chemistry 27】 or a salt thereof.

55. Formula (I) 【Chemistry 28】 [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R １ is C １～５ is alkyl, R ２ represents, independently at each occurrence, an optionally substituted C １０～３０ is alkyl, R ３ represents, independently at each occurrence, an optionally substituted C １０～３０ alkyl] or a salt thereof, Methods for making peptides or peptidomimetics.

56. 56. The method of claim 55, wherein the compound of formula (I) is used as a support for producing a peptide or peptidomimetic.

57. The peptidomimetic has the formula 【Chemistry 29】 or a salt thereof.

58. The peptidomimetic has the formula 【Transformation 30】 or a salt thereof.

59. Formula (I) 【Chemistry 31】 [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R １ is C １～５ is alkyl, R ２ is, for each occurrence, optionally substituted C １０～３０ independently selected from alkyl, R ３ is, for each occurrence, optionally substituted C １０～３０ alkyl] or a salt thereof.

60. The compound has the formula 【Chemistry 32】 60. The compound of claim 59, having the formula:

61. The compound has the formula 【Transformation 33】 60. The compound of claim 59, having the formula:

62. 61. A pharmaceutical composition comprising a peptidomimetic according to claim 57 or a compound according to any one of claims 59-60.

63. 61. A pharmaceutical dosage form comprising a peptidomimetic according to claim 57 or a compound according to any one of claims 59 to 60.

64. 61. A method of selectively targeting GLP1R on the cell surface using a peptidomimetic according to claim 57 or a compound according to any one of claims 59-60.

65. 65. The method of claim 64, wherein the cell is a mammalian cell.

66. 66. The method of claim 64 or 65, wherein the cell is a human cell.

67. A method for enhancing the activity of GLP1R in an individual in need thereof, comprising administering to the individual an effective amount of a peptidomimetic described in claim 57 or a compound described in any one of claims 59 to 60, a composition described in claim 62, or a dosage form described in claim 63.

68. A method for lowering blood glucose levels in an individual in need thereof, comprising administering to the individual an effective amount of a peptide mimetic described in claim 57 or a compound described in any one of claims 59 to 60, a composition described in claim 62, or a dosage form described in claim 63.

69. A method for reducing body weight in an individual in need thereof, comprising administering to the individual an effective amount of a peptidomimetic described in claim 57 or a compound described in any one of claims 59 to 60, a composition described in claim 62, or a dosage form described in claim 63.

70. A method for treating a GLP1R-related disease in an individual in need thereof, comprising administering to the individual an effective amount of a peptide mimetic described in claim 57 or a compound described in any one of claims 59 to 60, a composition described in claim 62, or a dosage form described in claim 63.

71. 71. The method of claim 70, wherein the GLP1R-related disease is type II diabetes, obesity, liver disease, coronary artery disease, or kidney disease.

72. 71. The method of claim 70, wherein the GLP1R-related disease is type II diabetes and / or obesity.

73. 73. The method of any one of claims 67 to 72, wherein the peptidomimetic of claim 57 or the compound of any one of claims 59 to 60, the composition of claim 62, or the dosage form of claim 63 is administered subcutaneously, intravenously, intradermally, intraperitoneally, or intramuscularly.

74. 1. A method of making a peptide or peptidomimetic comprising: (a) Formula (I) 【Transformation 34】 [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R １ is C １～５ is alkyl, R ２ is, for each occurrence, optionally substituted C １０～３０ independently selected from alkyl, R ３ is, for each occurrence, optionally substituted C １０～３０ alkyl] or a salt thereof, (b) providing a first amino acid (AA1), and (c) coupling said first amino acid (AA1) with a compound of formula (I) to form a peptide bond between said first amino acid (AA1) and the compound of formula (I); A method comprising:

75. 75. The method of claim 74, wherein the first amino acid (AA1) contains a protected amino group.

76. The first amino acid (AA1) is 【Chemistry 35】 75. The method of claim 74, wherein:

77. 75. The method of claim 74, further comprising activating a chemical group on the first amino acid (AA1) to prepare the first amino acid (AA1) for coupling with a compound of formula (I) prior to step (c).

78. (d) removing the protecting group from the first amino acid (AA1); (e) providing a second amino acid (AA2), and (f) coupling said second amino acid (AA2) with said first amino acid (I-AA1) to form a peptide bond (I-AA1-AA2) between said second amino acid (AA2) and said first amino acid; 75. The method of claim 74, further comprising:

79. the second amino acid (AA2) is 【Transformation 36】 79. The method of claim 78, wherein:

80. (g) removing the protecting group from the second amino acid (AA2); (h) providing a third amino acid (AA3), and (i) coupling said third amino acid (AA3) with said second amino acid to form a peptide bond (I-AA1-AA2-AA3) between said third amino acid (AA3) and said second amino acid; 79. The method of claim 78, further comprising:

81. the third amino acid (AA3) is 【Chemistry 37】 81. The method of claim 80, wherein:

82. (j) removing the protecting group from the third amino acid (AA3); (k) providing a fourth amino acid (AA4), and (l) coupling the fourth amino acid (AA4) with the third amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4) between the fourth amino acid (AA4) and the third amino acid; 81. The method of claim 80, further comprising:

83. the fourth amino acid (AA4) is 【Transformation 38】 83. The method of claim 82, wherein:

84. (m) removing the protecting group from the fourth amino acid (AA4); (n) providing a fifth amino acid (AA5), and (o) coupling the fifth amino acid (AA5) with the fourth amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5) between the fifth amino acid (AA5) and the fourth amino acid; 83. The method of claim 82, further comprising:

85. the fifth amino acid (AA5) is 【Chemistry 39】 85. The method of claim 84, wherein:

86. (p) removing the protecting group from the fifth amino acid (AA5); (q) providing a sixth amino acid (AA6), and (r) coupling the sixth amino acid (AA6) with the fifth amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5-AA6) between the sixth amino acid (AA6) and the fifth amino acid.

85. The method of claim 84, further comprising:

87. the sixth amino acid (AA6) is 【Chemistry 40】 87. The method of claim 86, wherein:

88. (s) removing the protecting group from the sixth amino acid (AA6); (t) providing a seventh amino acid (AA7), and (u) coupling the seventh amino acid (AA6) with the sixth amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5-AA6-AA7) between the seventh amino acid (AA7) and the sixth amino acid; 87. The method of claim 86, further comprising:

89. the seventh amino acid (AA7) is 【Chemistry 41】 89. The method of claim 88, wherein:

90. (v) removing the protecting group from the seventh amino acid (AA7); (w) providing an eighth amino acid (AA8), and (x) coupling the eighth amino acid (AA8) with the seventh amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5-AA6-AA7-AA8) between the eighth amino acid (AA8) and the seventh amino acid; 89. The method of claim 88, further comprising:

91. the eighth amino acid (AA8) is 【Chemistry 42】 91. The method of claim 90, wherein:

92. (y) removing the protecting group from the eighth amino acid (AA8); (z) providing a ninth amino acid (AA9), and (aa) coupling the ninth amino acid (AA9) with the eighth amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5-AA6-AA7-AA8-AA9) between the ninth amino acid (AA9) and the eighth amino acid; 91. The method of claim 90, further comprising:

93. the ninth amino acid (AA9) is 【Chemistry 43】 93. The method of claim 92, wherein:

94. (bb) removing the protecting group from the ninth amino acid (AA9); (cc) providing a tenth amino acid (AA10), and (dd) coupling the tenth amino acid (AA10) with the ninth amino acid to form a peptide bond (I-AA1-AA2-AA3-AA4-AA5-AA6-AA7-AA8-AA9-AA10) between the tenth amino acid (AA10) and the ninth amino acid.

93. The method of claim 92, further comprising:

95. The tenth amino acid (AA10) is 【Chemistry 44】 95. The method of claim 94, wherein:

96. (i) Amino acids (AA ｎ ) removing the protecting group from (ii) the subsequent amino acid (AA ｎ＋１ ), and (iii) the subsequent amino acid (AA ｎ＋１ ) to the amino acid (AA ｎ ) to form the subsequent amino acid (AA ｎ＋１ ) and the amino acid (AA ｎ forming a peptide bond between and repeating the steps of:

95. The method of claim 94, wherein the repeating step is performed 1 to 100 times.

97. (ee) removing the protecting group from the tenth amino acid (AA10); (ff) Formula (B) 【Chemistry 45】 wherein x is an integer from 1 to 15; and (gg) coupling a compound of formula (B) to one of the amino acids in said peptide or said peptidomimetic to form a triazole ring.

95. The method of claim 94, further comprising:

98. The compound of formula (B) is represented by the following formula: 【Chemistry 46】 98. The method of claim 97, comprising:

99. 99. The method of any one of claims 74 to 98, further comprising (hh) cleaving the peptide bond between the first amino acid (AA1) and the compound of formula (I) to obtain a peptide or peptidomimetic.

100. The first amino acid (AA1), the second amino acid (AA2), the third amino acid (AA3), the fourth amino acid (AA4), the fifth amino acid (AA5), the sixth amino acid (AA6), the seventh amino acid (AA7), the eighth amino acid (AA8), the ninth amino acid (AA9), the tenth amino acid (AA10), and any subsequent amino acid (AA ｎ＋１ ) Each one of them is, 【Chemistry 47】 [In the formula, PG ５ 99. The method of any one of claims 74 to 98, wherein each of the groups is independently selected from the group consisting of:

101. PG ５ is independently selected from the group consisting of triphenylmethyl (Trt), allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

102. The first amino acid (AA1), the second amino acid (AA2), the third amino acid (AA3), the fourth amino acid (AA4), the fifth amino acid (AA5), the sixth amino acid (AA6), the seventh amino acid (AA7), the eighth amino acid (AA8), the ninth amino acid (AA9), the tenth amino acid (AA10), and any subsequent amino acid (AA ｎ＋１ ) Each one of them is, 【Chemistry 48】 99. The method of any one of claims 74 to 98, wherein the hydroxyl group is independently selected from the group consisting of:

103. The peptidomimetic has the formula 【Chemistry 49】 or a salt thereof.

104. The peptidomimetic has the formula [Transformation 50] or a salt thereof.

105. Formula (AI) 【Chemistry 51】 [In the formula, m and n are independently integers from 0 to 5; R １ａ is C １～５ is alkyl, R ２ａ represents, independently at each occurrence, an optionally substituted C １０～３０ is alkyl, R ３ａ represents, independently at each occurrence, an optionally substituted C １０～３０ alkyl] A compound having the structure: or a salt thereof.

106. R １ａ is methyl.

107. R ２ａ is, independently at each occurrence, optionally replaced by C １８～２６ alkyl, and R ３ａ is, independently at each occurrence, optionally replaced by C １８～２６ 106. The compound of claim 105, which is alkyl.

108. Each R ２ａ are independently optionally substituted C ２２ alkyl, and each R ３ａ are independently optionally substituted C ２２ 106. The compound of claim 105, which is alkyl.

109. Formula (BI) 【Chemistry 52】 [In the formula, m is an integer from 1 to 5; R １ｂ is C １～５ is alkyl, R ２ｂ represents, independently at each occurrence, an optionally substituted C １０～３０ alkyl] A compound having the structure: or a salt thereof.

110. R １ｂ is methyl.

111. Each R ２ｂ are independently optionally substituted C １８～２６ 110. The compound of claim 109, which is alkyl.

112. Each R ２ｂ are independently optionally substituted C ２２ 110. The compound of claim 109, which is alkyl.

113. Formula (CI) 【Chemistry 53】 [In the formula, m and n are independently integers from 1 to 5, and (m+n)≧3; R ２ｃ represents, independently at each occurrence, an optionally substituted C １０～３０ is alkyl, R ３ｃ represents, independently at each occurrence, an optionally substituted C １０～３０ alkyl] A compound having the structure: or a salt thereof.

114. R ２ｃ is, independently at each occurrence, optionally replaced by C １８～２６ alkyl, and R ３ｃ is, independently at each occurrence, optionally replaced by C １８～２６ 114. The compound of claim 113, which is alkyl.

115. Each R ２ｃ are independently optionally substituted C ２２ alkyl, and each R ３ｃ are independently optionally substituted C ２２ 114. The compound of claim 113, which is alkyl.

116. Formula (DI) 【Chemistry 54】 [In the formula, k is an integer from 1 to 15, q is an integer from 0 to 10; R １ｄ is C １～６ is alkyl, PG ６ is a suitable protecting group, PG ７ is a suitable protecting group. or a salt thereof, comprising: (a) Structure 【Transformation 55】 providing a compound of formula (DII) having the formula: (b) forming a compound of formula (DI) from a compound of formula (DII). A process involving:

117. PG ６ is selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

118. PG ７ is selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

119. The compound of formula (DI) is represented by formula (DIa) or formula (DIb) 【Transformation 56】 117. The process of claim 116, having the structure:

120. The compound of formula (DII) is represented by the formula (DIIa) or the formula (DIIb) 【Chemistry 57】 117. The process of claim 116, having the structure:

121. wherein step (b) of forming a compound of formula (DI) comprises The compound of formula (DII) is reacted with a compound of formula (DIII) 【Chemistry 58】 with a compound of formula (II) 117. The process of claim 116, comprising:

122. The compound of formula (DIII) is a compound of formula (DIIIa) 【Chemistry 59】 122. The process of claim 121, having the structure:

123. structure 【Transformation 60】 or a salt thereof; and Before step (a), forming a compound of formula (DII) from a compound of formula (DIV).

117. The process of claim 116, further comprising:

124. The compound of formula (DIV) is a compound of formula (DIVa) or formula (DIVb) 【Chemistry 61】 124. The process of claim 123, having the structure:

125. said step of forming a compound of formula (DII) The compound of formula (DIV) is reacted with a compound of formula (DVa) PG ６ -X(DVa) or Formula (DVb) PG ６ ２ O(DVb) [In the formula, X is OSu, OTf, Cl, or Br, Producing a compound of formula (DII) 124. The process of claim 123, comprising:

126. The compound of formula (DVa) or formula (DVb) is AllocCl, Alloc ２ O, Cbz ２ O, CbzCl, FmocCl, FmocOSu, and Boc ２ 126. The process of claim 125, wherein the compound is selected from the group consisting of: O.

127. structure 【Transformation 62】 [In the formula, PG ８ is a suitable protecting group; and Forming a compound of formula (DIV) from a compound of formula (DVI) 124. The process of claim 123, further comprising:

128. said step of forming a compound of formula (DIV) reacting a compound of formula (DVI) with a deprotecting agent to form a compound of formula (DIV).

128. The process of claim 127, comprising:

129. The deprotecting agent is Pd(PPh) ３ , PhSiH ３ , H ２ 129. The process of claim 128, wherein the amine is selected from the group consisting of piperidine and trifluoroacetic acid (TFA).

130. The compound of formula (DVI) is a compound of formula (DVIa) or formula (DVIb) 【Transformation 63】 128. The process of claim 127, having the structure:

131. structure 【Chemistry 64】 providing a compound of formula (DVII) having the formula: Forming a compound of formula (DVI) from a compound of formula (DVII) 128. The process of claim 127, further comprising:

132. said step of forming a compound of formula (DVI) reacting a compound of formula (DVII) with a base to produce a compound of formula (DVI) 132. The process of claim 131, comprising:

133. 129. The process of claim 128, wherein the base is selected from the group consisting of LiOH, NaOH, and KOH.

134. The compound of formula (DVII) is represented by the formula (DVIIa) or the formula (DVIIb) 【Transformation 65】 132. The process of claim 131, having the structure:

135. structure 【Chemical Formula 66】 [In the formula, and LG′ is a suitable living group; and Forming a compound of formula (DVII) from a compound of formula (DVIII) 132. The process of claim 131, further comprising:

136. said step of forming a compound of formula (DVII) reacting the compound of formula (DVIII) with an azide group introducing agent to produce a compound of formula (DVII).

136. The process of claim 135, comprising:

137. The azide group introducing agent is NaN ３ , TMSN ３ , (PhO) ２ P(O)N ３ , Zn(N ３ ) ２ ＊ 2Py, and n-Bu ４ N.N. ３ 137. The process of claim 136, selected from the group consisting of:

138. 136. The process of claim 135, wherein LG' is selected from the group consisting of halogen, OTf, OMs, and OTs.

139. The compound of formula (DVIII) is a compound of formula (DVIIIa) or formula (DVIIIb) 【Transformation 67】 136. The process of claim 135, having the structure:

140. structure 【Transformation 68】 [In the formula, wherein LG″ is a suitable living group; and Forming a compound of formula (DVIII) from a compound of formula (DIX) 136. The process of claim 135, further comprising:

141. 141. The process of claim 140, wherein LG" is selected from the group consisting of halogen, OTf, OMs, and OTs.

142. said step of forming a compound of formula (DVIII) The compound of formula (DIX) is reacted with a compound of formula (DX) 【Transformation 69】 to produce a compound of formula (DVII) 141. The process of claim 140, comprising:

143. The compound of formula (DIX) is represented by the formula (DIXa) 【Transformation 70】 141. The process of claim 140, having the structure:

144. The compound of formula (DX) is represented by the formula (DXa) 【Chemistry 71】 143. The process of claim 142, having the structure:

145. structure 【Chemistry 72】 [In the formula, R ２ｄ is C １～６ wherein R is an alkylene; and Forming a compound of formula (DIX) from a compound of formula (DXI) 141. The process of claim 140, further comprising:

146. said step of forming a compound of formula (DIX) reacting the compound of formula (DXI) with a reducing agent to produce a compound of formula (DIX).

146. The process of claim 145, comprising:

147. The reducing agent is H ２ , N.H. ４ HCO ２ ,NABH(Oac) ３ , and LiAlH ４ 146. The process of claim 145, selected from the group consisting of:

148. The compound of formula (DXI) may be prepared by reacting a compound of formula (DXIa) 【Transformation 73】 142. The process of claim 141, having the structure:

149. structure 【Chemistry 74】 providing a compound of formula (DXII) having the formula: Forming a compound of formula (DXI) from a compound of formula (DXII) 146. The process of claim 145, further comprising:

150. said step of forming a compound of formula (DXI) The compound of formula (DXII) is reacted with a compound of formula (DXIII) PPh ３ ＊ C １～６ Alkyl-Hal(DXIII) [In the formula, wherein Hal is a halogen, Producing a compound of formula (DXI) 150. The process of claim 149, comprising:

151. The compound of formula (DXII) may be prepared by reacting a compound of formula (DXIIa) 【Chemistry 75】 150. The process of claim 149, having the structure:

152. The compound of formula (DXIII) is a compound of formula (DXIIIa) PPh ３ ＊ MeBr(DXIIIa) 151. The process of claim 150, having the structure:

153. structure 【Transformation 76】 providing a compound of formula (DXIV) having the formula: Forming a compound of formula (DXII) from a compound of formula (DXIV) 150. The process of claim 149, further comprising:

154. said step of forming a compound of formula (DXII) The compound of formula (DXIV) is reacted with a compound of formula (DXV) 【Chemical 77】 [In the formula, wherein LG"' is a suitable living group, Producing a compound of formula (DXII) 154. The process of claim 153, comprising:

155. 155. The process of claim 154, wherein LG"' is selected from the group consisting of halogen, OTf, OMs, and OTs.

156. The compound of formula (DXIV) is a compound of formula (DXIVa) 【Transformation 78】 154. The process of claim 153, having the structure:

157. structure 【Transformation 79】 [In the formula, LG ＊ is a suitable living group; and Forming a compound of formula (DX) from a compound of formula (DXVI) 143. The process of claim 142, further comprising:

158. The LG ＊ 158. The process of claim 157, wherein is selected from the group consisting of halogen, OTf, OMs, and OTs.

159. said step of forming a compound of formula (DX) Compound of formula (DXVI) with compound of formula (DXVII) 【Chemistry 80】 with a compound of formula (DX) 158. The process of claim 157, comprising:

160. The compound of formula (DXVI) may be prepared by reacting a compound of formula (DXVIc) 【Chemistry 81】 158. The process of claim 157, having the structure:

161. structure 【Chemistry 82】 providing a compound of formula (DXVIII) having the formula: Forming a compound of formula (DXVI) from a compound of formula (DXVIII) 158. The process of claim 157, further comprising:

162. said step of forming a compound of formula (DXVI) The compound of formula (DXVIII) is reacted with a compound of formula (DXIXa) LG ＊ -X(DXIXa) or formula (DXIXb) LG ＊ ２ O(DXIXb) [In the formula, X is OSu, Cl, or Br, Producing a compound of formula (DXVI) 162. The process of claim 161, comprising:

163. The compound of formula (DXIXa) or formula (DXIXb) is TfCl, Tf ２ O, MsCl, and Ms ２ 163. The process of claim 162, wherein the compound is selected from the group consisting of: O.

164. Formula (DII) 【Chemistry 83】 [In the formula, q is an integer from 0 to 10; R １ｄ is C １～６ is alkyl, PG ６ is a suitable protecting group. or a salt thereof, comprising: (a) Structure 【Chemical 84】 or a salt thereof; and (b) forming a compound of formula (DII) from a compound of formula (DXXI). A process involving:

165. PG ６ is selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

166. The compound of formula (DXXI) is the compound 【Chemical 85】 The process of claim 164, wherein the process is carried out by

167. 165. The process of claim 164, wherein the compound of formula (DXXI) is treated with a catalyst.

168. The catalyst is Pd(dppf)Cl ２ - DCM.

169. 165. The process of claim 164, wherein the compound of formula (DXXI) is treated with a base.

170. The base is K ２ CO ３ 170. The process of claim 169, wherein

171. The preparation is ２ 165. The process of claim 164, carried out in a solvent mixture consisting of O / iPrOH.

172. The compound of formula (DII) is represented by the formula (DIIa) or the formula (DIIb) 【Chemical 86】 165. The process of claim 164, having the structure:

173. The compound of formula (DXXI) 【Chemistry 87】 of 【Chemical 88】 165. The process of claim 164, wherein the compound is formed by exposing the compound to

174. The compound of formula (DXXI) is PPh ３ The process of claim 173, wherein the compound is formed in the presence of

175. 174. The process of claim 173, wherein the compound of formula (DXXI) is formed in the presence of DIAD.

176. 174. The process of claim 173, wherein the compound of formula (DXXI) is formed in the presence of 4 Å molecular sieves.

177. 174. The process of claim 173, wherein the compound of formula (DXXI) is formed in an organic solvent.

178. 178. The process of claim 177, wherein the organic solvent is THF. 【Request Item 179】 【Chemistry 89】 but, 【Chemistry 90】 PBin ２ 174. The process of claim 173, wherein the compound is formed by exposing the compound to [Request Item 180] [Chemistry 91] Pd(OAc) ２ 180. The process of claim 179, wherein the compound is formed in the presence of 【Request Item 181】 【Chemistry 92】 But PCy ３ 180. The process of claim 179, wherein the compound is formed in the presence of [Request Item 182] [Chemistry 93] 180. The process of claim 179, wherein is formed in the presence of KOAc. [Request Item 183] [Chemistry 94] 180. The process of claim 179, wherein is formed in the presence of dioxane. [Request Item 184] [Chemistry 95] 180. The process of claim 179, wherein is formed at an elevated temperature over a period of time. [Request Item 185] [Chemistry 96] but, 【Chemistry 97】 Bu ４ N ＋ Br ３ － 180. The process of claim 179, wherein the compound is formed by exposing the compound to 【Request Item 186】 【Chemistry 98】 186. The process of claim 185, wherein is formed in an organic solvent.

187. 187. The process of claim 186, wherein the organic solvent is a mixture of DCM and methanol.

188. Formula (DII) 【Chem.99】 [In the formula, q is an integer from 0 to 10; R １ｄ is C １～６ is alkyl, PG ６ is a suitable protecting group. or a salt thereof, comprising: (a) Structure 【Chemistry 100】 or a salt thereof; and (b) forming a compound of formula (DII) from a compound of formula (DIV). A process involving:

189. PG ６ is selected from the group consisting of allyloxycarbonyl (Alloc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc).

190. The compound of formula (DII) is represented by the formula (DIIa) or the formula (DIIb) 【Chemistry 101】 189. The process of claim 188, having the structure:

191. The compound of formula (DIV) is a compound of formula (DIVa) or formula (DIVb) 【Chemical Engineering 102】

192. said step of forming a compound of formula (DII) The compound of formula (DIV) is reacted with a compound of formula (DVa) PG ６ -X(DVa) or Formula (DVb) PG ６ ２ O(DVb) [In the formula, X is OSu, Otf, Cl, or Br, Producing a compound of formula (DII) 189. The process of claim 188, comprising:

193. The compound of formula (DVa) or formula (DVb) is AllocCl, Alloc ２ O, Cbz ２ O, CbzCl, FmocCl, FmocOSu, and Boc ２ 193. The process of claim 192, wherein the compound is selected from the group consisting of: O.

194. structure 【Chemistry 103】 [In the formula, PG ８ is a suitable protecting group; and Forming a compound of formula (DIV) from a compound of formula (DXVI) 189. The process of claim 188, further comprising:

195. said step of forming a compound of formula (DIV) reacting a compound of formula (DXVI) with a deprotecting agent to produce a compound of formula (DIV).

195. The process of claim 194, comprising:

196. The deprotecting agent is HCl, Pd(PPh) ３ , PhSiH ３ , H ２ 196. The process of claim 195, wherein the amine is selected from the group consisting of piperidine and trifluoroacetic acid (TFA).

197. The compound of formula (DXVI) is represented by formula (DXVIa) or formula (DXVIb) 【Chemical 104】 195. The process of claim 194, having the structure:

198. structure 【Chemistry 105】 [In the formula, and LG′ is a suitable living group; and Forming a compound of formula (DXVI) from a compound of formula (DXVII) 195. The process of claim 194, further comprising:

199. said step of forming a compound of formula (DXVI) reacting the compound of formula (DXVII) with an azide group introducing agent to produce a compound of formula (DXVI).

200. The process of claim 198, comprising:

200. The azide group introducing agent is NaN ３ , TMSN ３ , (PhO) ２ P(O)N ３ , Zn(N ３ ) ２ ＊ 2Py, and n-Bu ４ N.N. ３ 200. The process of claim 199, selected from the group consisting of:

201. 200. The process of claim 198, wherein LG' is selected from the group consisting of halogen, OTf, OMs, and OTs.

202. The compound of formula (DXVII) is represented by the formula (DXVIIa) or the formula (DXVIIb) 【Chemistry 106】 200. The process of claim 198, having the structure:

203. structure 【Chemistry 107】 [In the formula, wherein LG″ is a suitable living group; and Forming a compound of formula (DXVII) from a compound of formula (DIX) 200. The process of claim 198, further comprising:

204. 204. The process of claim 203, wherein LG" is selected from the group consisting of halogen, OTf, OMs, and OTs.

205. said step of forming a compound of formula (DXVII) The compound of formula (DIX) is reacted with a compound of formula (DXVIII) 【Chemistry 108】 to produce a compound of formula (DXVII) 204. The process of claim 203, comprising:

206. The compound of formula (DIX) is represented by the formula (DIXa) 【Chemistry 109】 The process of claim 203, having the structure:

207. The compound of formula (DXVIII) is a compound of formula (DXVIIIa) 【Chemical 110】 The process of claim 205, having the structure:

208. structure 【Chemistry 111】 providing a compound of formula (DXIX) having the formula: forming a compound of formula (DIX) from a compound of formula (DXIX) 204. The process of claim 203, further comprising:

209. said step of forming a compound of formula (DIX) The compound of formula (DXIX) is reacted with a compound of formula (DXV) 【Chemistry 112】 [In the formula, wherein LG"' is a suitable living group, Producing a compound of formula (DXIX) 209. The process of claim 208, comprising:

210. 210. The process of claim 209, wherein LG"' is selected from the group consisting of halogen, OTf, OMs, and OTs.

211. The compound of formula (DXIX) may be prepared by reacting a compound of formula (DXIXa) 【Chemistry 113】 The process of claim 208, having the structure:

212. The compound of formula (DXV) may be a compound of formula (DXVa) 【Chemical 114】 210. The process of claim 209, having the structure:

213. structure 【Chemical 115】 providing a compound of formula (DXX) having the formula: Forming a compound of formula (DXIX) from a compound of formula (DXX) 209. The process of claim 208, further comprising:

214. said step of forming a compound of formula (DXIX) reacting the compound of formula (DXX) with a LG″ introducing agent to produce a compound of formula (DXIX).

214. The process of claim 213, comprising:

215. The LG" introducing agent is NBu ４ Br ３ 215. The process of claim 214, wherein:

216. The compound of formula (DXX) may be a compound of formula (DXXa) 【Chemistry 116】 214. The process of claim 213, having the structure:

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