Peptides for incretin synthesis

Abstract

Crystalline forms of peptide fragments, methods for their preparation, and their use for preparing peptides are provided. The crystalline compounds of the present invention can be utilized as intermediates with improved purity and physical properties for peptide synthesis.

Description

[Technical Field]

[0001] (Related Applications) This application claims priority to U.S. Provisional Patent Application No. 63 / 378,397, filed October 5, 2022, and U.S. Provisional Patent Application No. 63 / 477,742, filed December 29, 2022, the entire contents of which are incorporated herein by reference.

[0002] (Sequence Listing) This application contains a Sequence Listing that has been submitted electronically in XML format, and is incorporated herein by reference in its entirety. The XML copy created on October 3, 2023 is named 30267_WO.XML and is 51,063 bytes in size.

[0003] FIELD OF THE INVENTION FIELD OF THE DISCLOSURE The present disclosure relates to peptides useful in the preparation of biopharmaceutical products, and methods for their preparation and use. [Background technology]

[0004] Diabetes mellitus is a chronic disease characterized by hyperglycemia due to defects in insulin secretion, insulin action, or both. In type 2 diabetes mellitus ("T2D"), the combined effects of impaired insulin secretion and insulin resistance are associated with elevated blood glucose levels.

[0005] Tirzepatide ("TZP"), a GIP / GLP1 dual agonist, is described and claimed in U.S. Patent No. 9,474,780. Tirzepatide may be useful for treating type 2 diabetes ("T2D"). Additional methods for preparing tirzepatide and intermediates in those processes are described in U.S. Patent Application Publication No. 2022 / 0135639(A1). Retatrutide ("GGG"), a GLP-1 / GIP / glucagon triple receptor agonist, is described in International Patent Publication Nos. 2019 / 125938 and 2021 / 034815. Mazdutide ("OXM"), a GLP-1 / glucagon dual agonist, is described and claimed in U.S. Patent No. 9,935,335 and International Patent Publication No. 2021 / 252829. Both retatortide and mazdutide may also be useful for treating diabetes and promoting weight loss. These patents and patent publications are incorporated herein by reference in their entireties.

[0006] Additional processes and intermediates are needed that enable improved technologies for the streamlined production of peptides such as tirzepatide, retatortide, and mazdutide with commercially desirable purity and volume. Similarly, efficient, environmentally friendly processes, including stable intermediates to provide peptides and other compounds with fewer purification steps, are needed. Improved environmentally friendly processes are also needed to provide peptide manufacturing processes that minimize waste streams to enhance both environmental and operator safety. Large-scale preparation of pharmaceutically acceptable peptides presents several technical challenges that can affect overall yield and purity. Processes are needed that avoid the use of transition metals and / or harsh reaction conditions that are incompatible with peptide synthesis.

[0007] The present disclosure seeks to meet these needs by providing novel intermediates and processes useful in the production of peptides such as tirzepatide, retatortide, and mazdutide.

[0008] Tirzepatide ("TZP") has the structure of SEQ ID NO:1, as shown below.

[0009] [ka]

[0010] Retatortide ("GGG") has the structure of SEQ ID NO: 12, as shown below.

[0011] [ka]

[0012] Mazdoutide ("OXM") has the structure of SEQ ID NO: 16, as shown below.

[0013] [ka] Summary of the Invention

[0014] The present disclosure describes peptide compounds useful for preparing tirzepatide (SEQ ID NO: 1), retatortide (SEQ ID NO: 12), mazdutide (SEQ ID NO: 16), and other peptides with improved efficacy.

[0015] In some embodiments, the present disclosure provides a compound of formula (I), or a salt, solvate, or hydrate thereof:

[0016] [ka] In the formula, R 1 is H or a protecting group, and R 2 is H or a protecting group, and R 3 is H or a protecting group, and R 4 is H or a protecting group, and R 1 , R 2 , R 3 , and R 4In some embodiments, R 1 is H, Fmoc, or Boc. In some embodiments, R 2 is H or t-butyl. In some embodiments, R 4 is H or benzyl. In some embodiments, R 3 is H, t-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, or trityl. 1 , R 2 , R 3 , and R 4 At least two of R 1 , R 2 , R 3 , and R 4 At least two of are protecting groups.

[0017] In some embodiments, the compound of formula (I) has the formula:

[0018] [ka] is.

[0019] In some embodiments, the compound of formula (I) has the formula:

[0020] [ka] is.

[0021] In some embodiments, the compound of formula (I) has the formula:

[0022] [ka] is.

[0023] In some embodiments, the compound of formula (I) has the formula:

[0024] [ka] is.

[0025] In some embodiments, the compound of formula (I) has the formula:

[0026] [ka] is.

[0027] In some embodiments, the compound of Formula (I) is a solvate. In some embodiments, the solvate is formed from methyl t-butyl ether (MTBE) or a mixture containing MTBE.

[0028] In some embodiments, the compound of Formula (I) is crystalline. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle of 5.2° 2θ ± 0.2 degrees, combined with one or more peaks selected from 8.4°, 8.8°, 10.4°, 15.5°, 17.1°, and 17.7°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle of 6.1° 2θ ± 0.2 degrees, combined with one or more peaks selected from 10.3°, 14.9°, 16.8°, 18.1°, and 18.2°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle of 6.2° 2θ ± 0.2 degrees, combined with one or more peaks selected from 8.9°, 12.3°, 14.9°, 15.4°, and 21.8°.

[0029] In some embodiments, the present disclosure provides a method of synthesizing a compound of formula (I), comprising: -Formula (Y 保護 )

[0030] [ka] In the formula, R 1 and R 2 is a protecting group, 保護 ) and a compound of formula (Aib 保護 )

[0031] [ka] In the formula, R 5 is a protecting group, 保護 ) to react with a compound of formula (Y 保護 -Aib 保護 forming a compound of

[0032] [ka] -R 5 and removing the protecting group of the formula (Y 保護 -Aib),

[0033] [ka] -Formula (Y 保護 -Aib) and a compound of formula (E 保護 )

[0034] [ka] In the formula, R 3 is a protecting group, 保護 ) to react with a compound of formula (Y 保護 -Aib-E 保護 forming a compound of

[0035] [ka] -Formula (Y 保護 -Aib-E 保護 ) and a compound of formula (G 保護 )

[0036] [ka] In the formula, R 4 is a protecting group, 保護 ) to react with a compound of formula (Y 保護 -Aib-E 保護 -G 保護 forming a compound of

[0037] [ka] and, optionally, -Protecting group R 1 , R 2 , R 3 , and R 4 removing one or more of:

[0038] In some embodiments, the present disclosure describes a method for synthesizing a polypeptide of SEQ ID NO: 1, comprising conjugating a compound of formula (I) to the N-terminus of a polypeptide of SEQ ID NO: 2 via the C-terminus of the compound.

[0039] In some embodiments, the present disclosure describes a method for synthesizing a polypeptide of SEQ ID NO: 26, comprising conjugating a compound of formula (I) via the C-terminus of the compound to the N-terminus of a polypeptide of SEQ ID NO: 27.

[0040] In some embodiments, the present disclosure provides a compound of formula (II), or a salt, solvate, or hydrate thereof:

[0041] [ka] In the formula, R 6 is H or a protecting group, and R 7 is H or a protecting group, and R 8 is H or a protecting group, and R 9 is H or a protecting group, and R 10is H or a protecting group, or a salt, solvate, or hydrate thereof. In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups. In some embodiments, R 6 is H, Fmoc, or Boc. In some embodiments, R 7 is H or t-butyl. In some embodiments, R 8 is H or t-butyl. In some embodiments, R 9 is H or t-butyl. In some embodiments, R 10 is H or benzyl. In some embodiments, R 6 , R 7 , R 8 , R 9 , and R 10 At least one of R is a protecting group. 6 , R 7 , R 8 , R 9 , and R 10 is H.

[0042] In some embodiments, the compound of formula (II) has the formula:

[0043] [ka] is a compound of

[0044] In some embodiments, the compound of Formula (I) is a solvate. In some embodiments, the solvate is formed from a solvent comprising heptane.

[0045] In some embodiments, the compound of Formula (I) is crystalline. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 7.7 to 7.9 degrees ± 0.2 degrees, combined with one or more peaks selected from 5.8°, 10.0°, 10.8-10.9°, 11.3-11.4°, 12.0-12.1°, 12.8°, 14.2-14.4°, and 16.8-17.0°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 7.3 degrees ± 0.2 degrees, combined with one or more peaks selected from 5.1°, 5.7°, 7.6°, 9.5°, and 12.4°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 7.8° ± 0.2 degrees in combination with one or more peaks selected from 8.5°, 11.5°, 12.0°, 12.8°, 14.3°, 15.5°, 20.2°, and 23.3°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 8.2 to 8.3° ± 0.2 degrees in combination with one or more peaks selected from 5.9°, 7.7°, 9.2°, 10.2°, 11.3°, 13.8 to 13.9°, 15.5 to 15.7°, 17.1°, and 18.5°.

[0046] In some embodiments, the present disclosure describes a method for synthesizing a polypeptide of SEQ ID NO: 1, comprising: (i) conjugating a compound of formula (II) via the N-terminus of the compound to the C-terminus of a polypeptide of SEQ ID NO: 3 to form a polypeptide of SEQ ID NO: 4; and (ii) conjugating the polypeptide of SEQ ID NO: 4 via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 5.

[0047] In some embodiments, the present disclosure provides a compound of formula (III), or a salt, solvate, or hydrate thereof:

[0048] [ka] In the formula, R 11 is H or a protecting group, and R 12 is H or a protecting group, or a salt, solvate, or hydrate thereof. In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups. In some embodiments, R 11 is H or t-butyl. In some embodiments, R 12 is H or t-butyl. In some embodiments, R 11 and R 12 At least one of R is a protecting group. 11 and R 12 is H.

[0049] In some embodiments, the compound of formula (III) has the formula:

[0050] [ka] is a compound of

[0051] In some embodiments, the compound of formula (III) is a solvate.

[0052] In some embodiments, the compound of Formula (III) is crystalline. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 8.0° ± 0.2 degrees, combined with one or more peaks selected from 7.0°, 10.3°, 14.1°, 15.2°, 16.7°, 18.0°, 19.0°, 19.7°, 20.8°, and 21.9°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 9.0° ± 0.2 degrees, combined with one or more peaks selected from 5.7°, 9.9°, 16.2°, 17.1°, 17.9°, 18.1°, 18.4°, 18.8°, 19.9°, 20.1°, and 22.5°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle of 10.5° 2θ ± 0.2 degrees in combination with one or more peaks selected from 5.6°, 11.9°, 13.3°, 15.4°, 15.6°, 18.1°, 19.9°, and 21.1°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle of 10.7° 2θ ± 0.2 degrees in combination with one or more peaks selected from 4.9°, 14.8°, 20.3°, and 21.5°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle of 10.5° 2θ ± 0.2 degrees in combination with one or more peaks selected from 5.9°, 10.5°, 10.9°, 12.1°, 13.1°, 15.9°, 17.5°, 20.9°, 21.1°, and 21.9°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle of 7.8° 2θ ± 0.2 degrees in combination with one or more peaks selected from 11.3°, 11.5°, 15.4°, 15.6°, and 21.5°.In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle of 10.0° ± 0.2 degrees 2θ in combination with one or more peaks selected from 8.1°, 12.5°, 13.5°, 14.7°, 17.8°, 18.8°, 20.0°, and 22.4°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle of 21.1° ± 0.2 degrees 2θ in combination with one or more peaks selected from 5.6°, 10.5°, 10.8°, 11.9°, 15.4°, and 23.8°.

[0053] In some embodiments, the present disclosure describes a method for synthesizing a polypeptide of SEQ ID NO: 1, comprising: (i) conjugating a compound of formula (III) via the N-terminus of the compound to the C-terminus of a polypeptide of SEQ ID NO: 6 to form a polypeptide of SEQ ID NO: 7; and (ii) conjugating the polypeptide of SEQ ID NO: 7 via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 8.

[0054] In some embodiments, the present disclosure provides a compound of formula (IV), or a salt, solvate, or hydrate thereof:

[0055] [ka] In the formula, R 13 is H or a protecting group, and R 13* is H or a protecting group, and R 14 is H or a protecting group, and R 15 is H or a protecting group, or a salt, solvate, or hydrate thereof. In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups. In some embodiments, R 13 is H, Fmoc, or Boc. In some embodiments, R 13* is H. In some embodiments, R 14 is H or t-butyl. In some embodiments, R15 is H or t-butyl. In some embodiments, R 13 , R 13* , R 14 , and R 15 At least one of R is a protecting group. 13 , R 13* , R 14 , and R 15 is H.

[0056] In some embodiments, the compound of formula (IV) has the formula:

[0057] [ka] is a compound of

[0058] In some embodiments, the compound of formula (IV) is a solvate.

[0059] In some embodiments, the compound of Formula (IV) is crystalline. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle of 5.1° 2θ ± 0.2 degrees in combination with one or more peaks selected from 4.3°, 6.1°, 8.0°, 10.1°, and 18.7°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle of 5.2° 2θ ± 0.2 degrees in combination with one or more peaks selected from 6.0°, 6.7°, 10.0°, 10.3°, 16.4°, 17.8°, 18.3°, 19.4°, and 22.4°.

[0060] In some embodiments, the present disclosure describes a method for synthesizing a polypeptide of SEQ ID NO: 1, comprising: (i) conjugating a compound of formula (IV) via the N-terminus of the compound to the C-terminus of a polypeptide of SEQ ID NO: 9 to form a polypeptide of SEQ ID NO: 7; and (ii) conjugating the polypeptide of SEQ ID NO: 7 via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 8.

[0061] In some embodiments, the compound of formula (IV) has the formula

[0062] [ka] is a compound of In the formula, R 54 is H or a protecting group, and R 55 is H or a protecting group, and R 56 is H or a protecting group. In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups. In some embodiments, R 54 is H or tert-butyl. In some embodiments, R 55 is H or tert-butyl. In some embodiments, R 56 is H or Boc. In some embodiments, R 54 , R 55 , and R 56 At least one of R 54 , R 55 , and R 56 At least one of is a protecting group.

[0063] In some embodiments, the compound of formula (IV) has formula (IV-b):

[0064] [ka] It has the following structure.

[0065] In some embodiments, the compound of formula (IV-b) is a solvate. In some embodiments, the compound is a solvate formed from acetone. In some embodiments, the compound is a desolvate.

[0066] In some embodiments, the compound of Formula (IV-b) is crystalline. In some embodiments, the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 5.8° and 18.5° ±0.2 degrees, in combination with one or more peaks selected from 8.6°, 9.4°, 12.9°, 13.8°, 17.2°, and 19.4°. In some embodiments, the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 7.0-7.1° and 7.5-7.7° ±0.2 degrees, in combination with one or more peaks selected from 5.3-5.4°, 9.7-9.9°, and 14.7-14.9°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 8.3° ± 0.2 degrees, combined with one or more peaks selected from 6.3°, 11.4°, 14.3°, and 16.6°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.2° ± 0.2 degrees, combined with one or more peaks selected from 6.8°, 8.6°, 15.8°, and 18.9°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 6.1° ± 0.2 degrees, combined with one or more peaks selected from 6.3°, 7.8°, 10.0°, and 12.4°.

[0067] In some embodiments, the present disclosure provides a method of synthesizing a compound of formula (IV-b): - a compound of formula (Fmoc-G) and a compound of formula (P 保護 ) to react with a compound of the formula (Fmoc-GP 保護 forming a compound of

[0068] [ka] (In the formula, R 57 is a protecting group) -Protecting group R 57to form the compound (Fmoc-GP),

[0069] [ka] - a compound of formula (Fmoc-GP) and a compound of formula (S 保護2 ) to form a compound of the formula (Fmoc-GPS 保護2 forming a compound of

[0070] [ka] (In the formula, R 54 and R 58 is a protecting group), -Protecting group R 58 (Fmoc-GPS 保護1 forming a compound of

[0071] [ka] -Formula(Fmoc-GPS 保護1 ) and compounds of formula (S 保護2 ) to form a compound of the formula (Fmoc-GPS 保護1 -S 保護2 forming a compound of

[0072] [ka] (In the formula, R 55 and R 59 is a protecting group) -Protecting group R 59 (Fmoc-FG 保護1 -S 保護1 -S 保護1 forming a compound of

[0073] [ka] -Formula(Fmoc-GPS 保護1 -S 保護1) and compounds of formula (G 保護 ) to form a compound of the formula (Fmoc-GPS 保護1 -S 保護1 -G 保護 )

[0074] [ka] forming a compound of (In the formula, R 60 is a protecting group) -Formula(Fmoc-GPS 保護1 -S 保護1 -G 保護 37. A compound of formula (I) above,

[0075] [ka] and optionally one or more protecting groups R 54 , R 55 , and R 56 A method is described, comprising removing

[0076] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0077] In some embodiments, the present disclosure describes a method for synthesizing a polypeptide of SEQ ID NO: 12, comprising: (i) conjugating a compound of formula (IV-b) via the C-terminus of the compound to the N-terminus of a polypeptide of SEQ ID NO: 22; and (ii) conjugating the resulting compound via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO: 23.

[0078] In some embodiments, the present disclosure describes a method for synthesizing a polypeptide of SEQ ID NO: 24, comprising conjugating a compound of formula (IV-b) to the N-terminus of a polypeptide of SEQ ID NO: 22 via the C-terminus of the compound.

[0079] In some embodiments, the present disclosure describes a method for synthesizing a polypeptide of SEQ ID NO: 16, comprising conjugating a compound of formula (IV-b) to the C-terminus of a polypeptide of SEQ ID NO: 25 via the N-terminus of the compound.

[0080] In some embodiments, the present disclosure describes a method for synthesizing a polypeptide of SEQ ID NO: 1, comprising: (i) conjugating a compound of formula (IV-b) via the C-terminus of the compound to the N-terminus of a polypeptide of SEQ ID NO: 8; and (ii) conjugating the resulting compound via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO: 9.

[0081] In some embodiments, the present disclosure provides a compound of formula (V), or a salt, solvate, or hydrate thereof:

[0082] [ka] In the formula, R 16 is H or a protecting group, and R 17 is H or a protecting group, and R 18 is H or a protecting group, and R 19 is H or a protecting group, and R 20 is H or a protecting group, and R 21 is H or a protecting group, or a salt, solvate, or hydrate thereof. In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups. In some embodiments, R 16 is H, Fmoc, or Boc. In some embodiments, R 17 is H or t-butyl. In some embodiments, R 18 is H or t-butyl. In some embodiments, R 19 is H or t-butyl. In some embodiments, R 20 is H or t-butyl. In some embodiments, R 21is H or benzyl. In some embodiments, R 16 , R 17 , R 18 , R 19 , R 20 , and R 21 At least one of R is a protecting group. 16 , R 17 , R 18 , R 19 , R 20 , and R 21 is H.

[0083] In some embodiments, the compound of formula (V) has the formula:

[0084] [ka] is a compound of

[0085] In some embodiments, the compound of Formula (V) is a solvate.

[0086] In some embodiments, the compound of Formula (V) is crystalline. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.8 to 6.1° ± 0.2 degrees, in combination with one or more peaks selected from 6.7 to 7.1° and 8.8 to 9.0°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.0 to 5.2° ± 0.2 degrees, in combination with one or more peaks selected from 5.3 to 5.4°, 5.7 to 6.0°, 6.1 to 6.2°, 7.6 to 7.9°, and 8.7 to 9.1°.

[0087] In some embodiments, the present disclosure describes a method for synthesizing a polypeptide of SEQ ID NO: 1, comprising: (i) conjugating a compound of formula (V) via the N-terminus of the compound to the C-terminus of a polypeptide of SEQ ID NO: 3 to form a polypeptide of SEQ ID NO: 10; and (ii) conjugating the polypeptide of SEQ ID NO: 10 via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 11.

[0088] In some embodiments, the present disclosure provides a compound of formula (VI), or a salt, solvate, or hydrate thereof:

[0089] [ka] In the formula, R 22 is H or a protecting group, and R 23 is H or a protecting group, and R 24 is H or a protecting group, and R 25 is H or a protecting group, or a salt, solvate, or hydrate thereof. In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups. In some embodiments, R 22 is H or Boc. In some embodiments, R 23 is H or tert-butyl. In some embodiments, R 24 is H or trityl. In some embodiments, R 25 is H or tert-butyl. In some embodiments, R 22 , R 23 , R 24 , and R 25 At least one of R 22 , R 23 , R 24 , and R 25 At least one of is a protecting group.

[0090] In some embodiments, the compound of formula (VI) has formula (VI-a):

[0091] [ka] is a compound of

[0092] In some embodiments, the compound of formula (VI) is a solvate. In some embodiments, the compound is a solvate formed from pentyl acetate, a mixture containing pentyl acetate, ethyl acetate, or a mixture containing 2-methyltetrahydrofuran and t-amyl methyl ether. In some embodiments, the mixture containing pentyl acetate is selected from a mixture containing pentyl acetate and t-butyl ethyl ether, a mixture containing pentyl acetate and t-amyl methyl ether, or a mixture containing pentyl acetate and heptane. In some embodiments, the compound is a desolvate or a partially desolvate.

[0093] In some embodiments, the compound of Formula (VI) is crystalline. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 6.3 to 6.4° ±0.2 degrees, in combination with one or more peaks selected from 4.5°, 7.1°, 13.0-13.1°, 15.9-16.0°, and 18.4-18.6°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.0 to 7.2° ±0.2 degrees, in combination with one or more peaks selected from 5.0-5.4°, 7.6-7.7°, 8.8-8.9°, 9.4-9.5°, and 12.5-12.7°.

[0094] In some embodiments, the present disclosure provides a method of synthesizing a compound of formula (VI): -Formula (Y 保護 ) and compounds of formula (Aib 保護 ) to react with a compound of formula (Y 保護 -Aib 保護 forming a compound of

[0095] [ka] (In the formula, R 22 , R 23 , and R 26 is a protecting group) -Protecting group R 26 By removing (Y 保護 -Aib),

[0096] [ka] -Formula (Y 保護 -Aib) and the compound of formula (Q 保護2 ) to react with a compound of formula (Y 保護 -Aib-Q 保護2 forming a compound of

[0097] [ka] (In the formula, R 24 and R 27 is a protecting group) -Protecting group R 27 By removing (Y 保護 -Aib-Q 保護1 forming a compound of

[0098] [ka] -Formula (Y 保護 -Aib-Q 保護1 ) and compounds of formula (G 保護 ) to form a compound of formula (VI),

[0099] [ka] (In the formula, R 25 is a protecting group), and optionally one or more protecting groups R 22 , R 23 , R 24 , and R25 A method is described, comprising removing

[0100] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0101] In some embodiments, the present disclosure describes a method for synthesizing a polypeptide of SEQ ID NO: 12, comprising conjugating a compound of formula (VI) to the N-terminus of a polypeptide of SEQ ID NO: 13 via the C-terminus of the compound.

[0102] In some embodiments, the present disclosure describes a method for synthesizing a polypeptide of SEQ ID NO: 14, comprising conjugating a compound of formula (VI) to the N-terminus of a polypeptide of SEQ ID NO: 15 via the C-terminus of the compound.

[0103] In some embodiments, the present disclosure provides a compound of formula (VII), or a salt, solvate, or hydrate thereof:

[0104] [ka] In the formula, R 28 is H or a protecting group, and R 29 is H or a protecting group, and R 30 is H or a protecting group, or a salt, solvate, or hydrate thereof. In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups. In some embodiments, R 28 is H or Boc. In some embodiments, R 29 is H or trityl. In some embodiments, R 30 is H or tert-butyl. In some embodiments, R 28 , R 29 , and R 30 At least one of R 28 , R 29, and R 30 At least one of is a protecting group.

[0105] In some embodiments, the compound of formula (VII) has formula (VII-a):

[0106] [ka] is a compound of

[0107] In some embodiments, the compound is of Formula (VII) and is a solvate. In some embodiments, the compound is a solvate formed from any one of a mixture of acetonitrile and methyl tert-butyl ether, a mixture of nitromethane and methyl tert-butyl ether, a mixture of tetrahydrofuran and methyl tert-butyl ether, methyl acetate, and ethyl acetate.

[0108] In some embodiments, the compound of Formula (VII) is crystalline. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 4.8° ± 0.2 degrees, combined with one or more peaks selected from 5.6°, 6.2°, 14.8°, and 15.6°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.3° ± 0.2 degrees, combined with one or more peaks selected from 7.7°, 10.5°, 11.3°, 11.6°, and 14.4°. In some embodiments, the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 6.2° and 6.9° ± 0.2 degrees.

[0109] In some embodiments, the present disclosure provides a method of synthesizing a compound of formula (VII): -Formula(H(dnp) 保護 ) and compounds of formula (Aib 保護 ) to form a compound of formula (H(dnp) 保護-Aib 保護 forming a compound of

[0110] [ka] (In the formula, R 28 and R 31 is a protecting group) -Protecting group R 31 By removing (H(dnp) 保護 -Aib),

[0111] [ka] -Formula(H(dnp) 保護 -Aib) and the compound of formula (Q 保護2 ) to form a compound of formula (H(dnp) 保護 -Aib-Q 保護2 forming a compound of

[0112] [ka] (In the formula, R 29 and R 32 is a protecting group) -Protecting group R 32 By removing (H(dnp) 保護 -Aib-Q 保護1 forming a compound of

[0113] [ka] -Formula(H(dnp) 保護 -Aib-Q 保護1 ) and compounds of formula (G 保護 with a compound of formula (VII) to form a compound of formula (VII),

[0114] [ka] (In the formula, R 30 is a protecting group), and optionally one or more protecting groups R 28 , R 29 , and R 30 A method is described, comprising removing

[0115] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0116] In some embodiments, the present disclosure describes a method of synthesizing a polypeptide of SEQ ID NO: 16, comprising conjugating a compound of formula (VII) to the N-terminus of a polypeptide of SEQ ID NO: 17 via the C-terminus of the compound.

[0117] In some embodiments, the present disclosure provides a compound of formula (VIII), or a salt, solvate, or hydrate thereof:

[0118] [ka] In the formula, R 33 is H or a protecting group, and R 34 is H or a protecting group, and R 35 is H or a protecting group, or a salt, solvate, or hydrate thereof. In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups. In some embodiments, R 33 is H or Boc. In some embodiments, R 34 is H or trityl. In some embodiments, R 35 is H or tert-butyl. In some embodiments, R 33 , R 34 , and R 35 At least one of R 33 , R 34 , and R 35 At least one of is a protecting group.

[0119] In some embodiments, the compound of Formula (VIII) has the formula (VIII-a):

[0120] [ka] is a compound of

[0121] In some embodiments, the compound of Formula (VIII) is a solvate. In some embodiments, the compound is a solvate formed from any one of a mixture of tetrahydrofuran and methyl tert-butyl ether, a mixture of tetrahydrofuran and heptane, a mixture of 1,4-dioxane and water, a mixture of ethyl acetate and methyl tert-butyl ether, and a mixture of acetonitrile and methyl tert-butyl ether.

[0122] In some embodiments, the compound of Formula (VIII) is crystalline. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 4.7° ± 0.2 degrees, combined with one or more peaks selected from 5.5°, 8.2°, 10.1°, 11.8°, 13.3°, 13.6°, and 18.9°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.8° ± 0.2 degrees, combined with one or more peaks selected from 5.3°, 8.9°, 9.2°, 15.2°, 18.6°, and 19.5°.

[0123] In some embodiments, the present disclosure provides a method of synthesizing a compound of formula (VIII): -Formula(H(trt) 保護 ) and compounds of formula (Aib 保護 ) to form a compound of formula (H(trt) 保護 -Aib 保護 forming a compound of

[0124] [ka] (In the formula, R 33 and R 36 is a protecting group) -Protecting group R 36 By removing (H(trt) 保護 -Aib),

[0125] [ka] -Formula(H(trt) 保護 -Aib) and the compound of formula (Q 保護2 ) to form a compound of formula (H(trt) 保護 -Aib-Q 保護2 forming a compound of

[0126] [ka] (In the formula, R 33 and R 37 is a protecting group) -Protecting group R 37 By removing (H(trt) 保護 -Aib-Q 保護1 forming a compound of

[0127] [ka] -Formula(H(trt) 保護 -Aib-Q 保護1 ) and compounds of formula (G 保護 to form a compound of formula (VIII),

[0128] [ka] (In the formula, R 35 is a protecting group), and optionally one or more protecting groups R 33 , R 34 , and R 35 A method is described, comprising removing

[0129] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0130] In some embodiments, the present disclosure describes a method for synthesizing a polypeptide of SEQ ID NO: 16, comprising conjugating a compound of formula (VIII) to the N-terminus of a polypeptide of SEQ ID NO: 17 via the C-terminus of the compound.

[0131] In some embodiments, the present disclosure provides a compound of formula (IX), or a salt, solvate, or hydrate thereof:

[0132] [ka] In the formula, R 38 is H or a protecting group, and R 39 is H or a protecting group, and R 40 is H or a protecting group, and R 41 is H or a protecting group, and R 42 is H or a protecting group, and R 43 is H or a protecting group, or a salt, solvate, or hydrate thereof. In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups. In some embodiments, R 38 is H or Fmoc. In some embodiments, R 39 is H or tert-butyl. In some embodiments, R 40 is H or tert-butyl. In some embodiments, R 41 is H or tert-butyl. In some embodiments, R 42 is H or Boc. In some embodiments, R 43 is H or tert-butyl. In some embodiments, R 38 , R 39 , R 40 , R 41 , R 42 , and R 43At least one of R 38 , R 39 , R 40 , R 41 , R 42 , and R 43 At least one of is a protecting group.

[0133] In some embodiments, the compound of formula (IX) has formula (IX-a):

[0134] [ka] is a compound of

[0135] In some embodiments, the compound is of Formula (IX) and is a solvate. In some embodiments, the compound is a solvate formed from any one of a mixture of methyl acetate and dibutyl ether, a mixture of acetone and dibutyl ether, a mixture of acetonitrile and dibutyl ether, a mixture of ethyl acetate and dibutyl ether, a mixture of methyl acetate and heptane, and a mixture of methyl ethyl ketone and dibutyl ether. In some embodiments, the compound is a desolvate or anhydrous.

[0136] In some embodiments, the compound of Formula (IX) is crystalline. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.3° ± 0.2 degrees, combined with one or more peaks selected from 6.0°, 6.9°, 7.2°, 8.0°, 12.2°, and 15.6°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.8° ± 0.2 degrees, combined with one or more peaks selected from 4.4°, 6.6°, 10.1°, 11.4°, 13.4°, and 15.5°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 4.5° and 5.5° ± 0.2 degrees, combined with one or more peaks selected from 6.0° and 7.3°.

[0137] In some embodiments, the present disclosure provides a method of synthesizing a compound of formula (IX): -Formula(D 保護 ) and a compound of formula (Y 保護2 ) to react with a compound of formula (D 保護 -Y 保護2 forming a compound of

[0138] [ka] (In the formula, R 38 , R 39 , R 40 , and R 44 is a protecting group) -Protecting group R 44 Remove (D 保護 -Y 保護1 forming a compound of

[0139] [ka] -Formula(D 保護 -Y 保護1 ) and compounds of formula (S 保護2 ) to react with a compound of formula (D保護 -Y 保護1 -S 保護2 forming a compound of

[0140] [ka] (In the formula, R 41 and R 45 is a protecting group) -Protecting group R 45 Remove (D 保護 -Y 保護1 -S 保護1 forming a compound of

[0141] [ka] -Formula(D 保護 -Y 保護1 -S 保護1 ) and compounds of formula (K 保護2 ) to form a compound of formula (IX),

[0142] [ka] (In the formula, R 42 and R 43 is a protecting group), and optionally one or more protecting groups R 38 , R 39 , R 40 , R 41 , R 42 , and R 43 A method is described, comprising removing

[0143] In some embodiments, the protecting group is selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0144] In some embodiments, the present disclosure describes a method of synthesizing a polypeptide of SEQ ID NO: 16, comprising: (i) conjugating a compound of formula (IX) via the C-terminus of the compound to the N-terminus of a polypeptide of SEQ ID NO: 18; and (ii) conjugating the compound via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO: 19.

[0145] In some embodiments, the present disclosure provides a compound of formula (X), or a salt, solvate, or hydrate thereof:

[0146] [ka] In the formula, R 46 is H or a protecting group, and R 47 is H or a protecting group, and R 48 is H or a protecting group, and R 49 is H or a protecting group, and R 50 is H or a protecting group, and R 51 is H or a protecting group, or a salt, solvate, or hydrate thereof. In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups. In some embodiments, R 46 is H or tert-butyl. In some embodiments, R 47 is H or tert-butyl. In some embodiments, R 48 is H or Boc. In some embodiments, R 49 is H or Fmoc. In some embodiments, R 50 is H or tert-butyl. In some embodiments, R 51 is H or tert-butyl. In some embodiments, R 46 , R 47 , R 48 , R 49 , R 50 , and R 51 At least one of R 46 , R 47 , R48 , R 49 , R 50 , and R 51 At least one of is a protecting group.

[0147] In some embodiments, the compound of Formula (X) has the formula (Xa):

[0148] [ka] is a compound of

[0149] In some embodiments, the compound of Formula (X) is a solvate. In some embodiments, the compound is a solvate formed from ethanol or isopropyl alcohol. In some embodiments, the compound is a desolvate.

[0150] In some embodiments, the compound of Formula (X) is crystalline. In some embodiments, the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 18.1° and 18.7° ±0.2 degrees, in combination with one or more peaks selected from 5.7°, 8.7°, 13.7°, 14.3°, 15.9°, and 16.2°. In some embodiments, the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 5.9° and 10.5° ±0.2 degrees, in combination with one or more peaks selected from 7.1°, 8.9°, 14.6°, and 16.6°. In some embodiments, the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 7.8° and 20.3° ±0.2 degrees, in combination with one or more peaks selected from 5.8°, 15.5°, and 19.5°. In some embodiments, the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 5.9° and 7.4° ±0.2 degrees, in combination with one or more peaks selected from 6.5°, 6.9°, and 14.8°.

[0151] In some embodiments, the present disclosure provides a method of synthesizing a compound of formula (X): -Formula (Y 保護 ) and compounds of formula (S 保護2 ) to react with a compound of formula (Y 保護 -S 保護2 forming a compound of

[0152] [ka] (In the formula, R 46 , R 47 , R 48 , and R 52 is a protecting group) -Protecting group R 52 By removing (Y 保護 -S 保護1 forming a compound of

[0153] [ka] -Formula (Y 保護 -S 保護1 ) and compounds of formula (K 保護2 ) to react with a compound of formula (Y 保護 -S 保護1 -K 保護2 forming a compound of

[0154] [ka] (In the formula, R 49 and R 53 is a protecting group) -Protecting group R 53 By removing (Y 保護 -S 保護1 -K 保護1 forming a compound of

[0155] [ka] -Formula (Y 保護 -S 保護1 -K 保護1 ) and a compound of formula (Y保護2 ) to form a compound of formula (X),

[0156] [ka] (In the formula, R 50 and R 51 is a protecting group) optionally one or more protecting groups R 46 , R 47 , R 48 , R 49 , R 50 , and R 51 A method is described, comprising removing

[0157] In some embodiments, the protecting group is selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0158] In some embodiments, the present disclosure describes a method of synthesizing a polypeptide of SEQ ID NO: 16, comprising: (i) conjugating a compound of formula (X) via the C-terminus of the compound to the N-terminus of a polypeptide of SEQ ID NO: 20; and (ii) conjugating the resulting compound via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO: 21.

[0159] In some embodiments, the present disclosure provides a compound of formula (XI), or a salt, solvate, or hydrate thereof:

[0160] [ka] In the formula, R 64 is H or a protecting group, and R 65 is H or a protecting group, and R 66 is H or a protecting group, or a salt, solvate, or hydrate thereof. In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups. In some embodiments, R 64 , R 65, and R 66 At least one of R 64 , R 65 , and R 66 At least one of is a protecting group.

[0161] In some embodiments, the compound of formula (XI) has the formula:

[0162] [ka] is a compound of

[0163] In some embodiments, the compound of formula (XI) is a solvate.

[0164] In some embodiments, the compound of Formula (XI) is crystalline. In some embodiments, the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 6.1° and 8.5° ±0.2 degrees, in combination with one or more peaks selected from 5.8°, 16.9°, 18.5°, 18.8°, 19.3°, and 20.9°.

[0165] In some embodiments, the present disclosure provides a compound of formula (XII), or a salt, solvate, or hydrate thereof:

[0166] [ka] In the formula, R 61 is H or a protecting group, and R 62 is H or a protecting group, and R 63 is H or a protecting group, or a salt, solvate, or hydrate thereof. In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups. In some embodiments, R 61 is H or tert-butyl. In some embodiments, R 62is H or tert-butyl. In some embodiments, R 63 is H or tert-butyl. In some embodiments, R 61 , R 62 , and R 63 At least one of R 61 , R 62 , and R 63 At least one of is a protecting group.

[0167] In some embodiments, the compound of formula (XII) has formula (XII-a):

[0168] [ka] is a compound of

[0169] In some embodiments, the compound of formula (XII) is a solvate. In some embodiments, the compound is a desolvate or anhydrous.

[0170] In some embodiments, the compound of Formula (XII) is crystalline. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 11.4° ±0.2 degrees, combined with one or more peaks selected from 6.0°, 8.9°, 12.7°, 13.6°, 14.6°, 17.0°, and 18.8°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 10.6° ±0.2 degrees, combined with one or more peaks selected from 7.1°, 12.1°, 13.6°, 14.2°, 15.2°, 16.0°, and 16.8°. In some embodiments, the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 10.1° and 15.5° ±0.2 degrees in combination with one or more peaks selected from 6.1°, 8.7°, 11.4°, 16.6°, and 19.2°.

[0171] In some embodiments, the present disclosure provides a compound of formula (XIII), or a salt, solvate, or hydrate thereof:

[0172] [ka] In the formula, R 67 is H or a protecting group, and R 68 is H or a protecting group, and R 69 is H or a protecting group, or a salt, solvate, or hydrate thereof. In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups. In some embodiments, R 67 is H or tert-butyl. In some embodiments, R 68 is H or tert-butyl. In some embodiments, R 69 is H or tert-butyl. In some embodiments, R 67 , R 68 , and R 69 At least one of R 67 , R 68 , and R 69 At least one of is a protecting group.

[0173] In some embodiments, the compound of formula (XIII) has the formula:

[0174] [ka] is a compound of

[0175] In some embodiments, the compound of formula (XIII) is a solvate. In some embodiments, the compound is a desolvate or anhydrous.

[0176] In some embodiments, the compound of Formula (XIII) is crystalline. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.0° ± 0.2 degrees, combined with one or more peaks selected from 8.3, 9.7, and 11.2°. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.2° ± 0.2 degrees, combined with one or more peaks selected from 5.3, 8.1, 14.4, and 16.2°.

[0177] In some embodiments, the present disclosure provides a compound of formula (XIV), or a salt, solvate, or hydrate thereof:

[0178] [ka] In the formula, R 70 is H or a protecting group, and R 71 is H or a protecting group, and R 72 is H or a protecting group, and R 73 is H or a protecting group, and R 74 is H or a protecting group, or a salt, solvate, or hydrate thereof. In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups. In some embodiments, R 70 is H, Fmoc, or tert-butyl. In some embodiments, R 71 is H or tert-butyl. In some embodiments, R 72 is H or tert-butyl. In some embodiments, R 73 is H or tert-butyl. In some embodiments, R 74 is H, Fmoc, or tert-butyl. In some embodiments, R 70 , R 71 , R 72 , R 73 , and R 74At least one of R 70 , R 71 , R 72 , R 73 , and R 74 At least one of is a protecting group.

[0179] In some embodiments, the compound of formula (XIV) has the formula:

[0180] [ka] is a compound of

[0181] In some embodiments, the compound of formula (XIV) is a solvate. In some embodiments, the compound is a desolvate or anhydrous.

[0182] In some embodiments, the compound of Formula (XIV) is crystalline. In some embodiments, the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.5°±0.2 degrees in combination with one or more peaks selected from 6.1, 8.7, 10.6, 15.0, 16.1, and 18.6°. [Brief explanation of the drawings]

[0183] [Figure 1A] Representative X-ray powder diffraction ("XRPD") patterns of Y-Aib-EG tetramer Form A, Form B, and Form C are shown, respectively. [Figure 1B] Representative X-ray powder diffraction ("XRPD") patterns of Y-Aib-EG tetramer Form A, Form B, and Form C are shown, respectively. [Figure 1C] Representative X-ray powder diffraction ("XRPD") patterns of Y-Aib-EG tetramer Form A, Form B, and Form C are shown, respectively. [Figure 2A] Representative XRPD patterns of TFTS tetramer Form A, Form B, Form C, and Form D are shown, respectively. [Figure 2B] Representative XRPD patterns of TFTS tetramer Form A, Form B, Form C, and Form D are shown, respectively. [Figure 2C] Representative XRPD patterns of TFTS tetramer Form A, Form B, Form C, and Form D are shown, respectively. [Figure 2D] Representative XRPD patterns of TFTS tetramer Form A, Form B, Form C, and Form D are shown, respectively. [Figure 3] 1 shows a polymorph map relating the various crystalline solid forms of PSSG-NH2. [Figure 4A] Representative XRPD patterns of PSSG-NH2 tetramer Form A, Form B, Form C, Form D, Form E, Form F, Form G, and Form H are shown, respectively. [Figure 4B] Representative XRPD patterns of PSSG-NH2 tetramer Form A, Form B, Form C, Form D, Form E, Form F, Form G, and Form H are shown, respectively. [Figure 4C] Representative XRPD patterns of PSSG-NH2 tetramer Form A, Form B, Form C, Form D, Form E, Form F, Form G, and Form H are shown, respectively. [Figure 4D] Representative XRPD patterns of PSSG-NH2 tetramer Form A, Form B, Form C, Form D, Form E, Form F, Form G, and Form H are shown, respectively. [Figure 4E] Representative XRPD patterns of PSSG-NH2 tetramer Form A, Form B, Form C, Form D, Form E, Form F, Form G, and Form H are shown, respectively. [Figure 4F] Representative XRPD patterns of PSSG-NH2 tetramer Form A, Form B, Form C, Form D, Form E, Form F, Form G, and Form H are shown, respectively. [Figure 4G]Representative XRPD patterns of PSSG-NH2 tetramer Form A, Form B, Form C, Form D, Form E, Form F, Form G, and Form H are shown, respectively. [Figure 4H] Representative XRPD patterns of PSSG-NH2 tetramer Form A, Form B, Form C, Form D, Form E, Form F, Form G, and Form H are shown, respectively. [Figure 5A] Representative XRPD patterns of GPSSG-NH2 pentamer Form A and Form B are shown, respectively. [Figure 5B] Representative XRPD patterns of GPSSG-NH2 pentamer Form A and Form B are shown, respectively. [Figure 6A] Representative XRPD patterns of TFTSD pentamer Form A and Form B, respectively, are shown. The traces in Figures 6A and 6B show the product results obtained in various experiments. [Figure 6B] Representative XRPD patterns of TFTSD pentamer Form A and Form B, respectively, are shown. The traces in Figures 6A and 6B show the product results obtained in various experiments. [Figure 7A] Representative XRPD patterns of Y-Aib-QG tetramer Form A and Form B are shown, respectively. [Figure 7B] Representative XRPD patterns of Y-Aib-QG tetramer Form A and Form B are shown, respectively. [Figure 8A] Representative XRPD patterns of H(dnp)-Aib-QG tetramer Form A, Form B, and Form C are shown, respectively. [Figure 8B] Representative XRPD patterns of H(dnp)-Aib-QG tetramer Form A, Form B, and Form C are shown, respectively. [Figure 8C] Representative XRPD patterns of H(dnp)-Aib-QG tetramer Form A, Form B, and Form C are shown, respectively. [Figure 9A] Representative XRPD patterns of H(trt)-Aib-QG tetramer Form A and Form B are shown, respectively. [Figure 9B]Representative XRPD patterns of H(trt)-Aib-QG tetramer Form A and Form B are shown, respectively. [Figure 10A] Representative XRPD patterns of DYSK tetramer Form A, Form B, and Form C are shown, respectively. [Figure 10B] Representative XRPD patterns of DYSK tetramer Form A, Form B, and Form C are shown, respectively. [Figure 10C] Representative XRPD patterns of DYSK tetramer Form A, Form B, and Form C are shown, respectively. [Figure 11] 1 shows a polymorphic map of the relationship between various crystalline solid forms of YSKY. [Figure 12A] Representative XRPD patterns of YSKY Form A, Form B, Form C, and Form D are shown, respectively. [Figure 12B] Representative XRPD patterns of YSKY Form A, Form B, Form C, and Form D are shown, respectively. [Figure 12C] Representative XRPD patterns of YSKY Form A, Form B, Form C, and Form D are shown, respectively. [Figure 12D] Representative XRPD patterns of YSKY Form A, Form B, Form C, and Form D are shown, respectively. [Figure 13] 1 shows a polymorphic map relating the various crystalline solid forms of Fmoc-GPSSG-NH2. [Figure 14A] Representative XRPD patterns of Fmoc-GPSSG-NH2 pentamer Form A, Form B, Form C, Form D, and Form E are shown, respectively. [Figure 14B] Representative XRPD patterns of Fmoc-GPSSG-NH2 pentamer Form A, Form B, Form C, Form D, and Form E are shown, respectively. [Figure 14C] Representative XRPD patterns of Fmoc-GPSSG-NH2 pentamer Form A, Form B, Form C, Form D, and Form E are shown, respectively. [Figure 14D]Representative XRPD patterns of Fmoc-GPSSG-NH2 pentamer Form A, Form B, Form C, Form D, and Form E are shown, respectively. [Figure 14E] Representative XRPD patterns of Fmoc-GPSSG-NH2 pentamer Form A, Form B, Form C, Form D, and Form E are shown, respectively. [Figure 15A] Representative XRPD patterns of Fmoc-GPSSG-OH tetramer Form A and Form B are shown, respectively. [Figure 15B] Representative XRPD patterns of Fmoc-GPSSG-OH tetramer Form A and Form B are shown, respectively. [Figure 16A] Representative XRPD patterns of GGG side chain Form A, Form B, and Form C are shown, respectively. [Figure 16B] Representative XRPD patterns of GGG side chain Form A, Form B, and Form C are shown, respectively. [Figure 16C] Representative XRPD patterns of GGG side chain Form A, Form B, and Form C are shown, respectively. [Figure 17A] 17A and 17B show representative XRPD patterns of HAPPPS-NH2HCl tetramer Form A and Form B, respectively. Figure 17B shows a representative XRPD pattern of HAPPPS-NH2HCl tetramer Form A in dry form. [Figure 17B] 17A and 17B show representative XRPD patterns of HAPPPS-NH2HCl tetramer Form A and Form B, respectively. Figure 17B shows a representative XRPD pattern of HAPPPS-NH2HCl tetramer Form A in dry form. [Figure 17C] 17A and 17B show representative XRPD patterns of HAPPPS-NH2HCl tetramer Form A and Form B, respectively. Figure 17B shows a representative XRPD pattern of HAPPPS-NH2HCl tetramer Form A in dry form. [Figure 18] A representative XRPD pattern of HA Fmoc-SSGAPPPS-NH2 tetramer form A is shown. [Figure 19]LCMS analysis results for coupling agent tests are shown. [Figure 20] 1 shows the UPLC-MS results for the Fmoc-Y-Aib-EG compound. DETAILED DESCRIPTION OF THE INVENTION

[0184] The present disclosure relates to crystalline forms of peptide compounds that have desirable physical properties, such as high purity and hygroscopicity, and are useful for preparing biological pharmaceutical compounds. The crystalline peptide compounds provided herein may be useful for preparing biological pharmaceutical peptides such as tirzepatide (SEQ ID NO: 1), retatortide (SEQ ID NO: 12), and mazduthide (SEQ ID NO: 16) with improved efficiency.

[0185] Formula (I) In one aspect, the present disclosure provides a compound of formula (I), or a salt, solvate, or hydrate thereof:

[0186] [ka] In the formula, R 1 is H or a protecting group, and R 2 is H or a protecting group, and R 3 is H or a protecting group, and R 4 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0187] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups.

[0188] In another aspect, the present disclosure provides a crystalline compound of formula (I), or a salt, solvate, or hydrate thereof,

[0189] [ka] In the formula, R 1is H or a protecting group, and R 2 is H or a protecting group, and R 3 is H or a protecting group, and R 4 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0190] In some embodiments, R 1 is H, Fmoc, or Boc. In some embodiments, R 2 is H or t-butyl. In some embodiments, R 3 is H or t-butyl. In some embodiments, R 4 is H or benzyl.

[0191] In some embodiments, R 1 , R 2 , R 3 and R 4 At least one of R is a protecting group. 1 , R 2 , R 3 , and R 4 is H.

[0192] In some embodiments, R 3 is tert-butyl. In some embodiments, R 3 is benzyl. In some embodiments, R 3 is carboxybenzyl. In some embodiments, R 3 is methyl. In some embodiments, R 3 is allyl. In some embodiments, R 3 is cyclohexyl. In some embodiments, R 3 is trityl.

[0193] In some embodiments, the crystalline form of Formula (I) is represented by Formula (Ia):

[0194] [ka] The compound has the structure: Boc-l-Tyr(tBu)-Aib-Glu(OtBu)-Gly-OH.

[0195] In some embodiments, the compound of Formula (I) or (Ia) is crystalline. In some embodiments, the compound of Formula (I) or (Ia) is a solvate. In some embodiments, the solvate of Formula (I) or (Ia) is crystalline. In some embodiments, the compound of Formula (I) or (Ia) is a solvate formed from methyl t-butyl ether (MTBE) or a mixture containing MTBE.

[0196] In some embodiments, the compound of Formula (Ia) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.2° ± 0.2 degrees in combination with one or more peaks selected from the group consisting of 8.4°, 8.8°, 10.4°, 15.5°, 17.1°, and 17.7°.

[0197] In some embodiments, the compound of Formula (Ia) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 6.1°±0.2 degrees in combination with one or more peaks selected from the group consisting of 10.3°, 14.9°, 16.8°, 18.1°, and 18.2°.

[0198] In some embodiments, the compound of Formula (Ia) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 6.2°±0.2 degrees in combination with one or more peaks selected from the group consisting of 8.9°, 12.3°, 14.9°, 15.4°, and 21.8°.

[0199] In some embodiments, the compound of Formula (Ia) is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta ±0.2 degrees of 5.2°, 8.4°, 8.8°, 10.4°, 15.5°, 17.1°, and 17.7°.

[0200] In some embodiments, the compound of formula (I) is

[0201] [ka] is.

[0202] In some embodiments, the compound of formula (I) is

[0203] [ka] is.

[0204] In some embodiments, the compound of formula (I) is

[0205] [ka] is.

[0206] In some embodiments, the compound of formula (I) is

[0207] [ka] is.

[0208] The present disclosure also provides a method of synthesizing a compound of formula (I), comprising: (i) Formula (Y 保護 )

[0209] [ka] In the formula, R 1 and R 2 is a protecting group, 保護 ) and a compound of formula (Aib 保護 )

[0210] [ka] In the formula, R 5is a protecting group, 保護 ) to react with a compound of formula (Y 保護 -Aib 保護 forming a compound of

[0211] [ka] R 5 and removing the protecting group of the formula (Y 保護 -Aib),

[0212] [ka] (ii) Formula (Y 保護 -Aib) and a compound of formula (E 保護 )

[0213] [ka] In the formula, R 3 is a protecting group, 保護 ) to react with a compound of formula (Y 保護 -Aib-E 保護 forming a compound of

[0214] [ka] (iii) Equation (Y 保護 -Aib-E 保護 ) and a compound of formula (G 保護 )

[0215] [ka] In the formula, R 4 is a protecting group, 保護 ) to react with a compound of formula (Y 保護 -Aib-E 保護 -G 保護 forming a compound of

[0216] [ka]

[0217] and optionally, (iv) a protecting group R 1 , R 2 , R 3 , and R 4 and removing one or more of:

[0218] In some embodiments, R 5 is t-butyl.

[0219] In some embodiments, R 1 is Fmoc and R 2 is t-butyl, and R 3 is t-butyl, and R 4 is t-butyl, and R 5 is t-butyl.

[0220] Formula (II) In another aspect, the present disclosure provides a compound of formula (II), or a salt, solvate, or hydrate thereof:

[0221] [ka] In the formula, R 6 is H or a protecting group, and R 7 is H or a protecting group, and R 8 is H or a protecting group, and R 9 is H or a protecting group, and R 10 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0222] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups. In some embodiments, the protecting group is Boc or Fmoc.

[0223] In some embodiments, R 6 is H, Fmoc, or Boc. In some embodiments, R 7 is H or t-butyl. In some embodiments, R 8 is H or t-butyl. In some embodiments, R 9 is H or t-butyl. In some embodiments, R 10 is H or benzyl. In some embodiments, R 6 , R 7 , R 8 , R 9 , and R 10 At least one of R is a protecting group. 6 , R 7 , R 8 , R 9 , and R 10 is H.

[0224] In some embodiments, formula (II) is represented by formula (II-a):

[0225] [ka] The compound has the structure: Fmoc-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-OH.

[0226] In some embodiments, the compound of Formula (II) or (II-a) is crystalline. In some embodiments, the compound of Formula (II) or (II-a) is a solvate. In some embodiments, the compound of Formula (II) or (II-a) is a solvate formed from a solvent comprising heptane. In some embodiments, the solvate of the compound of Formula (II) or (II-a) is crystalline.

[0227] In some embodiments, the compound of Formula (II-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 7.7 to 7.9° ± 0.2 degrees in combination with one or more peaks selected from the group consisting of 5.8°, 10.0°, 10.8-10.9°, 11.3-11.4°, 12.0-12.1°, 12.8°, 14.2-14.4°, and 16.8-17.0°.

[0228] In some embodiments, the compound of Formula (II-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 7.3° ± 0.2 degrees in combination with one or more peaks selected from the group consisting of 5.1°, 5.7°, 7.6°, 9.5°, and 12.4°.

[0229] In some embodiments, the compound of Formula (II-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 7.8° ± 0.2 degrees in combination with one or more peaks selected from the group consisting of 8.5°, 11.5°, 12.0°, 12.8°, 14.3°, 15.5°, 20.2°, and 23.3°.

[0230] In some embodiments, the compound of Formula (II-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 8.2 to 8.3° ±0.2 degrees in combination with one or more peaks selected from the group consisting of 5.9°, 7.7°, 9.2°, 10.2°, 11.3°, 13.8 to 13.9°, 15.5 to 15.7°, 17.1°, and 18.5°.

[0231] Formula (III) In another aspect, the present disclosure provides a compound of formula (III), or a salt, solvate, or hydrate thereof:

[0232] [ka] In the formula, R 11 is H or a protecting group, and R 12is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0233] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups.

[0234] In some embodiments, R 11 is H or t-butyl. In some embodiments, R 12 is H or t-butyl. In some embodiments, R 11 and R 12 At least one of R is a protecting group. 11 and R 12 is H.

[0235] In some embodiments, formula (III) is represented by formula (III-a):

[0236] [ka] The structure of the protease is Pro-Ser(tBu)-Ser(tBu)-Gly-NH2.

[0237] In some embodiments, the compound of Formula (III) or (III-a) is crystalline. In some embodiments, the compound of Formula (III) or (III-a) is a solvate. In some embodiments, the solvate form of Formula (III) or (III-a) is crystalline.

[0238] In some embodiments, the compound of Formula (III-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 8.0° ± 0.2 degrees in combination with one or more peaks selected from the group consisting of 7.0°, 10.3°, 14.1°, 15.2°, 16.7°, 18.0°, 19.0°, 19.7°, 20.8°, and 21.9°.

[0239] In some embodiments, the compound of Formula (III-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 9.0° ± 0.2 degrees in combination with one or more peaks selected from the group consisting of 5.7°, 9.9°, 16.2°, 17.1°, 17.9°, 18.1°, 18.4°, 18.8°, 19.9°, 20.1°, and 22.5°.

[0240] In some embodiments, the compound of Formula (III-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 10.5° ± 0.2 degrees in combination with one or more peaks selected from the group consisting of 5.6°, 11.9°, 13.3°, 15.4°, 15.6°, 18.1°, 19.9°, and 21.1°.

[0241] In some embodiments, the compound of Formula (III-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 10.7° ± 0.2 degrees in combination with one or more peaks selected from the group consisting of 4.9°, 14.8°, 20.3°, and 21.5°.

[0242] In some embodiments, the compound of Formula (III-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 10.5° ± 0.2 degrees in combination with one or more peaks selected from the group consisting of 5.9°, 10.5°, 10.9°, 12.1°, 13.1°, 15.9°, 17.5°, 20.9°, 21.1°, and 21.9°.

[0243] In some embodiments, the compound of Formula (III-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 7.8° ± 0.2 degrees in combination with one or more peaks selected from the group consisting of 11.3°, 11.5°, 15.4°, 15.6°, and 21.5°.

[0244] In some embodiments, the compound of Formula (III-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 10.0° ± 0.2 degrees in combination with one or more peaks selected from the group consisting of 8.1°, 12.5°, 13.5°, 14.7°, 17.8°, 18.8°, 20.0°, and 22.4°.

[0245] In some embodiments, the compound of Formula (III-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 21.1° ± 0.2 degrees in combination with one or more peaks selected from the group consisting of 5.6°, 10.5°, 10.8°, 11.9°, 15.4°, and 23.8°.

[0246] Formula (IV) In another aspect, the present disclosure provides a compound of formula (IV), or a salt, solvate, or hydrate thereof:

[0247] [ka] In the formula, R 13 is H or a protecting group, and R 13* is H or a protecting group, and R 14 is H or a protecting group, and R 15 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0248] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups.

[0249] In some embodiments, R 13 is H, Fmoc, or Boc. In some embodiments, R 13* is H. In some embodiments, R 14 is H or t-butyl. In some embodiments, R 15is H or t-butyl. In some embodiments, R 13 , R 14 , and R 15 At least one of R is a protecting group. 13 , R 13* , R 14 , and R 15 is H.

[0250] In some embodiments, formula (IV) is represented by formula (IV-a):

[0251] [ka] The amino acid sequence of the amino acid sequence of the nucleotide sequence of the amino acid ...

[0252] In some embodiments, the compound of Formula (IV) or (IV-a) is crystalline. In some embodiments, the compound of Formula (IV) or (IV-a) is a solvate. In some embodiments, the solvate of Formula (IV) or (IV-a) is crystalline.

[0253] In some embodiments, the compound of formula (IV-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.1° ± 0.2 degrees in combination with one or more peaks selected from the group consisting of 4.3°, 6.1°, 8.0°, 10.1°, and 18.7°.

[0254] In some embodiments, the compound of Formula (IV-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.2° ± 0.2 degrees in combination with one or more peaks selected from the group consisting of 6.0°, 6.7°, 10.0°, 10.3°, 16.4°, 17.8°, 18.3°, 19.4°, and 22.4°.

[0255] In some embodiments, Formula (IV) has the structure of Formula (IV-b):

[0256] [ka] In the formula, R 54 is H or a protecting group, and R 55 is H or a protecting group, and R 56 is H or a protecting group.

[0257] In some embodiments, the protecting group is selected from Boc, Fmoc, tert-butyl, and trityl groups. 54 is H or tert-butyl. In some embodiments, R 55 is H or tert-butyl. In some embodiments, R 56 is H or Boc. In some embodiments, R 54 , R 55 , and R 56 At least one of R 54 , R 55 , and R 56 At least one of is a protecting group.

[0258] In some embodiments, formula (IV) is Fmoc-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-NH2 having the structure of formula (IV-c):

[0259] [ka]

[0260] In some embodiments, the compound of formula (IV-c) is a solvate.

[0261] In some embodiments, the solvate is formed from acetone.

[0262] In some embodiments, the compound of formula (IV-c) is a desolvate.

[0263] In some embodiments, the compound of formula (IV-c) is crystalline.

[0264] In some embodiments, the compound of formula (IV-c) is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 5.8° and 18.5° ±0.2 degrees in combination with one or more peaks selected from 8.6°, 9.4°, 12.9°, 13.8°, 17.2°, and 19.4°.

[0265] In some embodiments, the compound of Formula (IV-c) is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta ±0.2 degrees of 7.0-7.1° and 7.5-7.7° in combination with one or more peaks selected from 5.3-5.4°, 9.7-9.9°, and 14.7-14.9°.

[0266] In some embodiments, the compound of formula (IV-c) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 8.3° ±0.2 degrees in combination with one or more peaks selected from 6.3°, 11.4°, 14.3°, and 16.6°.

[0267] In some embodiments, the compound of formula (IV-c) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.2° ±0.2 degrees in combination with one or more peaks selected from 6.8°, 8.6°, 15.8°, and 18.9°.

[0268] In some embodiments, the compound of formula (IV-c) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 6.1° ±0.2 degrees in combination with one or more peaks selected from 6.3°, 7.8°, 10.0°, and 12.4°.

[0269] The present disclosure also provides a method of synthesizing a compound of formula (IV-b), comprising: (i) Reacting a compound of formula (Fmoc-G) with a compound of formula (P保護 ) to react with a compound of the formula (Fmoc-GP 保護 forming a compound of

[0270] [ka] (In the formula, R 57 is a protecting group) (ii) Protecting group R 57 to form the compound (Fmoc-GP),

[0271] [ka] (iii) Reacting a compound of formula (Fmoc-GP) with a compound of formula (S 保護2 ) to form a compound of the formula (Fmoc-GPS 保護2 forming a compound of

[0272] [ka] (In the formula, R 54 and R 58 is a protecting group) (iv) Protecting group R 58 (Fmoc-GPS 保護1 forming a compound of

[0273] [ka] (v) Formula (Fmoc-GPS 保護1 ) and compounds of formula (S 保護2 ) to form a compound of the formula (Fmoc-GPS 保護1 -S 保護2 forming a compound of

[0274] [ka] (In the formula, R 55 and R 59 is a protecting group) (vi) Protecting group R 59 (Fmoc-FG 保護1 -S 保護1 -S 保護1 forming a compound of

[0275] [ka] (vii) Formula (Fmoc-GPS 保護1 -S 保護1 ) and compounds of formula (G 保護 ) to form a compound of the formula (Fmoc-GPS 保護1 -S 保護1 -G 保護 forming a compound of

[0276] [ka] (In the formula, R 60 is a protecting group) Equation (viii) (Fmoc-GPS 保護1 -S 保護1 -G 保護 converting a compound of formula (IV-b) into a compound of formula (IV-c),

[0277] [ka] (ix) optionally, one or more protecting groups R 54 , R 55 , and R 56 The method includes removing

[0278] In some embodiments, the protecting group is selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0279] The present disclosure also provides a method for synthesizing a polypeptide of SEQ ID NO: 12, comprising conjugating a compound of formula (IV-b) via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 22, and conjugating a compound of formula (IV-b) via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO: 23.

[0280] The present disclosure also provides a method for synthesizing a polypeptide of SEQ ID NO: 24, comprising conjugating a compound of formula (IV-b) via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 22.

[0281] The present disclosure also provides a method for synthesizing a polypeptide of SEQ ID NO: 16, comprising conjugating a compound of formula (IV-b) via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO: 25.

[0282] The present disclosure also provides a method for synthesizing a polypeptide of SEQ ID NO: 1, comprising conjugating a compound of formula (IV-b) via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 8, and conjugating a compound of formula (IV-b) via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO: 9.

[0283] Formula (V) In yet another aspect, the present disclosure provides a compound of formula (V), or a salt, solvate, or hydrate thereof:

[0284] [ka] In the formula, R 16 is H or a protecting group, and R 17 is H or a protecting group, and R 18 is H or a protecting group, and R 19 is H or a protecting group, and R 20 is H or a protecting group, and R 21 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0285] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups.

[0286] In some embodiments, R 16 is H, Fmoc, or Boc. In some embodiments, R 17 is H or t-butyl. In some embodiments, R 18 is H or t-butyl. In some embodiments, R 19 is H or t-butyl. In some embodiments, R 20 is H or t-butyl. In some embodiments, R 21 is H or benzyl. In some embodiments, R 16 , R 17 , R 18 , R 19 , R 20 , and R 21 At least one of R is a protecting group. 16 , R 17 , R 18 , R 19 , R 20 , and R 21 is H.

[0287] In some embodiments, formula (V) is formula (Va):

[0288] [ka] The compound has the structure: Fmoc-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-OH.

[0289] In some embodiments, the compound of Formula (V) or (Va) is crystalline.

[0290] In some embodiments, the compound of Formula (Va) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.8 to 6.1° ± 0.2 degrees, in combination with one or more peaks selected from the group consisting of 6.7 to 7.1° and 8.8 to 9.0°.

[0291] In some embodiments, the compound of Formula (Va) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.0 to 5.2° ± 0.2 degrees in combination with one or more peaks selected from 5.3 to 5.4°, 5.7 to 6.0°, 6.1 to 6.2°, 7.6 to 7.9°, and 8.7 to 9.1°.

[0292] In some embodiments, the polypeptide compounds described herein can be used to synthesize the active polypeptide TZP (SEQ ID NO: 1, 39 amino acids). In some embodiments, some of the compounds disclosed herein are conjugated with other polypeptide fragments to form full-length TZP. In some embodiments, the crystalline compounds of the present invention (e.g., the Y-Aib-EG tetramer of Formula (Ia)) can be used as pure, stable, and easy-to-use intermediates for synthesizing TZP in high yields. In some embodiments, crystallization of the compounds can help eliminate impurities from the crude material. Compared to amorphous crude material, crystalline compounds can exhibit improved purity (e.g., as measured by UPLC-MS analysis). In some embodiments, crystallization of crude compounds reduces the required solvent flow, providing a "greener," more ecologically desirable process. In some embodiments, crystallization of crude compounds spiked with 1% dimer (impurity) can result in crystalline compounds with nearly complete elimination of the dimer. In some embodiments, crystallization also improves physical properties, such as hygroscopicity. In some embodiments, crystalline compounds (such as the Y-Aib-EG tetramer of formula (Ia)) may exhibit significantly reduced weight gain due to moisture absorption compared to amorphous compounds.

[0293] A variety of synthetic routes can be selected using peptide synthesis techniques known in the art. In any given synthetic route, one or more compounds described in this disclosure can be used.

[0294] In some embodiments, a method of synthesizing a polypeptide of SEQ ID NO: 1 (TZP) is disclosed, comprising conjugating a compound of Formula (I) via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 2 (TZP amino acids 5-39).

[0295] Some embodiments disclose a method for synthesizing a polypeptide of SEQ ID NO: 26 (TZP 1-14 aa, which is Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Xaa Leu, where Xaa is Aib), comprising conjugating a compound of formula (I) via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 27 (TZP 5-14, which is Thr Phe Thr Ser Asp Tyr Ser Ile Xaa Leu, where Xaa is Aib).

[0296] Formula (VI) In another aspect, the present disclosure provides a compound of formula (VI), or a salt, solvate, or hydrate thereof:

[0297] [ka] In the formula, R 22 is H or a protecting group, and R 23 is H or a protecting group, and R 24 is H or a protecting group, and R 25 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0298] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups.22 is H or Boc. In some embodiments, R 23 is H or tert-butyl. In some embodiments, R 24 is H or trityl. In some embodiments, R 25 is H or tert-butyl. In some embodiments, R 22 , R 23 , R 24 , and R 25 At least one of R 22 , R 23 , R 24 , and R 25 At least one of is a protecting group.

[0299] In some embodiments, formula (VI) is represented by formula (VI-a):

[0300] [ka] The compound has the structure: Boc-Tyr(tBu)-Aib-Gln(trt)-Gly-OH.

[0301] In some embodiments, the compound of Formula (VI) or (VI-a) is a solvate. In some embodiments, the solvate is formed from pentyl acetate, a mixture containing pentyl acetate, ethyl acetate, or a mixture containing 2-methyltetrahydrofuran and t-amyl methyl ether.

[0302] In some embodiments, the mixture comprising pentyl acetate is selected from a mixture comprising pentyl acetate and t-butyl ethyl ether, a mixture comprising pentyl acetate and t-amyl methyl ether, or a mixture comprising pentyl acetate and heptane.

[0303] In some embodiments, the compound of formula (VI) or (VI-a) is a desolvate or a partial desolvate.

[0304] In some embodiments, the compound of formula (VI) or (VI-a) is crystalline.

[0305] In some embodiments, the compound of Formula (VI-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 6.3 to 6.4° ±0.2 degrees in combination with one or more peaks selected from 4.5°, 7.1°, 13.0-13.1°, 15.9-16.0°, and 18.4-18.6°.

[0306] In some embodiments, the compound of Formula (VI-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.0 to 7.2° ±0.2 degrees in combination with one or more peaks selected from 5.0 to 5.4°, 7.6 to 7.7°, 8.8 to 8.9°, 9.4 to 9.5°, and 12.5 to 12.7°.

[0307] The present disclosure also provides a method of synthesizing a compound of formula (VI), comprising: (i) Formula (Y 保護 ) and compounds of formula (Aib 保護 ) to react with a compound of formula (Y 保護 -Aib 保護 forming a compound of

[0308] [ka] (In the formula, R 22 , R 23 , and R 26 is a protecting group) (ii) Protecting group R 26 By removing (Y 保護 -Aib),

[0309] [ka] (iii) Equation (Y 保護 -Aib) and the compound of formula (Q 保護2 ) to react with a compound of formula (Y 保護-Aib-Q 保護2 forming a compound of

[0310] [ka] (In the formula, R 24 and R 27 is a protecting group) (iv) Protecting group R 27 By removing (Y 保護 -Aib-Q 保護1 forming a compound of

[0311] [ka] (v) Formula (Y 保護 -Aib-Q 保護1 ) and compounds of formula (G 保護 ) to form a compound of formula (VI),

[0312] [ka] (In the formula, R 25 is a protecting group), and (vi) optionally, one or more protecting groups R 22 , R 23 , R 24 , and R 25 The method includes removing

[0313] In some embodiments, the protecting group is selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0314] The present disclosure further provides a method of synthesizing a polypeptide of SEQ ID NO: 12, comprising conjugating the C-terminus of a compound of formula (VI-a) to the N-terminus of a polypeptide of SEQ ID NO: 13.

[0315] The present disclosure also provides a method for synthesizing a polypeptide of SEQ ID NO: 14, comprising conjugating the C-terminus of a compound of formula (VI-a) to the N-terminus of a polypeptide of SEQ ID NO: 15.

[0316] Formula (VII) In another aspect, the present disclosure provides a compound of formula (VII), or a salt, solvate, or hydrate thereof:

[0317] [ka] In the formula, R 28 is H or a protecting group, and R 29 is H or a protecting group, and R 30 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0318] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups. 28 is H or Boc. In some embodiments, R 29 is H or trityl. In some embodiments, R 30 is H or tert-butyl. In some embodiments, R 28 , R 29 , and R 30 At least one of R 28 , R 29 , and R 30 At least one of is a protecting group.

[0319] In some embodiments, Formula (VII) has Formula (VII-a):

[0320] [ka] The compound has the structure: Boc-Tyr(tBu)-Aib-Gln(trt)-Gly-OH.

[0321] In some embodiments, the compound of Formula (VII) or (VII-a) is a solvate, ie, a solvate formed from a mixture of acetonitrile and methyl tert-butyl ether, a mixture of nitromethane and methyl tert-butyl ether, a mixture of tetrahydrofuran and methyl tert-butyl ether, methyl acetate, and ethyl acetate.

[0322] In some embodiments, the compound of formula (VII) or (VII-a) is crystalline.

[0323] In some embodiments, the compound of Formula (VII) or (VII-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 4.8° ±0.2 degrees in combination with one or more peaks selected from 5.6°, 6.2°, 14.8°, and 15.6°.

[0324] In some embodiments, the compound of Formula (VII-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.3° ±0.2 degrees in combination with one or more peaks selected from 7.7°, 10.5°, 11.3°, 11.6°, and 14.4°.

[0325] In some embodiments, the compound of formula (VII-a) is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 6.2° and 6.9° ±0.2 degrees.

[0326] The present disclosure also provides a method of synthesizing a compound of formula (VII), comprising: (i) Equation (H(dnp) 保護 ) and compounds of formula (Aib 保護 ) to form a compound of formula (H(dnp) 保護 -Aib保護 forming a compound of

[0327] [ka] (In the formula, R 28 and R 31 is a protecting group) (ii) Protecting group R 31 By removing (H(dnp) 保護 -Aib),

[0328] [ka] (iii) Formula (H(dnp) 保護 -Aib) and the compound of formula (Q 保護2 ) to form a compound of formula (H(dnp) 保護 -Aib-Q 保護2 forming a compound of

[0329] [ka] (In the formula, R 29 and R 32 is a protecting group) (iv) Protecting group R 32 By removing (H(dnp) 保護 -Aib-Q 保護1 forming a compound of

[0330] [ka] (v) Formula (H(dnp) 保護 -Aib-Q 保護1 ) and compounds of formula (G 保護 with a compound of formula (VII) to form a compound of formula (VII),

[0331] [ka] (In the formula, R 30 is a protecting group), and (vi) optionally, one or more protecting groups R 28 , R 29 , and R 30 The method includes removing

[0332] The present disclosure also provides a method for synthesizing a polypeptide of SEQ ID NO: 16, comprising conjugating a compound of Formula (VII) or Formula (VII-a) via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 17.

[0333] Formula (VIII) In another aspect, the present disclosure provides a compound of formula (VIII), or a salt, solvate, or hydrate thereof:

[0334] [ka] In the formula, R 33 is H or a protecting group, and R 34 is H or a protecting group, and R 35 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0335] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups. 33 is H or Boc. In some embodiments, R 34 is H or trityl. In some embodiments, R 35 is H or tert-butyl. In some embodiments, R 33 , R 34 , and R 35 At least one of R 33 , R 34 , and R 35 At least one of is a protecting group.

[0336] In some embodiments, Formula (VIII) has Formula (VIII-a):

[0337] [ka] The compound has the structure: Boc-Tyr(tBu)-Aib-Gln(trt)-Gly-OH.

[0338] In some embodiments, the compound of Formula (VIII) or (VIII-a) is a solvate. In some embodiments, the solvate is formed from any one of a mixture of tetrahydrofuran and methyl tert-butyl ether, a mixture of tetrahydrofuran and heptane, a mixture of 1,4-dioxane and water, a mixture of ethyl acetate and methyl tert-butyl ether, and a mixture of acetonitrile and methyl tert-butyl ether.

[0339] In some embodiments, the compound of Formula (VIII) or (VIII-a) is crystalline.

[0340] In some embodiments, the compound of Formula (VIII-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 4.7° ±0.2 degrees in combination with one or more peaks selected from 5.5°, 8.2°, 10.1°, 11.8°, 13.3°, 13.6°, and 18.9°.

[0341] In some embodiments, the compound of Formula (VIII-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.8° ±0.2 degrees in combination with one or more peaks selected from 5.3°, 8.9°, 9.2°, 15.2°, 18.6°, and 19.5°.

[0342] The present disclosure also provides a method of synthesizing a compound of formula (VIII), comprising: (i) Formula (H(trt) 保護 ) and compounds of formula (Aib 保護) to form a compound of formula (H(trt) 保護 -Aib 保護 forming a compound of

[0343] [ka] (In the formula, R 33 and R 36 is a protecting group) (ii) Protecting group R 36 By removing (H(trt) 保護 -Aib),

[0344] [ka] (iii) Formula (H(trt) 保護 -Aib) and the compound of formula (Q 保護2 ) to form a compound of formula (H(trt) 保護 -Aib-Q 保護2 forming a compound of

[0345] [ka] (In the formula, R 33 and R 37 is a protecting group) (iv) Protecting group R 37 By removing (H(trt) 保護 -Aib-Q 保護1 forming a compound of

[0346] [ka] (v) Formula (H(trt) 保護 -Aib-Q 保護1 ) and compounds of formula (G 保護 to form a compound of formula (VIII),

[0347] [ka] (In the formula, R 35 is a protecting group), and (vi) optionally, one or more protecting groups R 33 , R 34 , and R 35 The method includes removing

[0348] In some embodiments, the protecting group is selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0349] The present disclosure also provides a method for synthesizing a polypeptide of SEQ ID NO: 16, comprising conjugating a compound of formula (VIII-a) via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 17.

[0350] Formula (IX) In another aspect, the present disclosure provides a compound of formula (IX), or a salt, solvate, or hydrate thereof:

[0351] [ka] In the formula, R 38 is H or a protecting group, and R 39 is H or a protecting group, and R 40 is H or a protecting group, and R 41 is H or a protecting group, and R 42 is H or a protecting group, and R 43 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0352] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups.

[0353] In some embodiments, R 38 is H or Fmoc. In some embodiments, R 39is H or tert-butyl. In some embodiments, R 40 is H or tert-butyl. In some embodiments, R 41 is H or tert-butyl. In some embodiments, R 42 is H or Boc. In some embodiments, R 43 is H or tert-butyl, and in some embodiments, R 38 , R 39 , R 40 , R 41 , R 42 , and R 43 At least one of R 38 , R 39 , R 40 , R 41 , R 42 , and R 43 At least one of is a protecting group.

[0354] In some embodiments, formula (IX) has formula (IX-a):

[0355] [ka] The compound has the structure: Boc-Tyr(tBu)-Aib-Gln(trt)-Gly-OH.

[0356] In some embodiments, the compound of formula (IX) or (IX-a) is a solvate.

[0357] In some embodiments, the solvate is formed from any one of a mixture of methyl acetate and dibutyl ether, a mixture of acetone and dibutyl ether, a mixture of acetonitrile and dibutyl ether, a mixture of ethyl acetate and dibutyl ether, a mixture of methyl acetate and heptane, and a mixture of methyl ethyl ketone and dibutyl ether.

[0358] In some embodiments, the compound of formula (IX) or (IX-a) is a desolvate or anhydrous.

[0359] In some embodiments, the compound of formula (IX-a) is crystalline.

[0360] In some embodiments, the compound of Formula (IX-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.3° ±0.2 degrees in combination with one or more peaks selected from 6.0°, 6.9°, 7.2°, 8.0°, 12.2°, and 15.6°.

[0361] In some embodiments, the compound of formula (IX-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.8° ±0.2 degrees in combination with one or more peaks selected from 4.4°, 6.6°, 10.1°, 11.4°, 13.4°, and 15.5°.

[0362] In some embodiments, the compound of formula (IX-a) is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 4.5° and 5.5° ±0.2 degrees, in combination with one or more peaks selected from 6.0° and 7.3°.

[0363] The present disclosure also provides a method of synthesizing a compound of formula (IX), comprising: (i) Formula (D 保護 ) and a compound of formula (Y 保護2 ) to react with a compound of formula (D 保護 -Y 保護2 forming a compound of

[0364] [ka] (In the formula, R 38 , R 39 , R 40 , and R 44 is a protecting group) (ii) Protecting group R44 Remove (D 保護 -Y 保護1 forming a compound of

[0365] [ka] (iii) Formula (D 保護 -Y 保護1 ) and compounds of formula (S 保護2 ) to react with a compound of formula (D 保護 -Y 保護1 -S 保護2 forming a compound of

[0366] [ka] (In the formula, R 41 and R 45 is a protecting group) (iv) Protecting group R 45 Remove (D 保護 -Y 保護1 -S 保護1 forming a compound of

[0367] [ka] (v) Formula (D 保護 -Y 保護1 -S 保護1 ) and compounds of formula (K 保護2 ) to form a compound of formula (IX),

[0368] [ka] (In the formula, R 42 and R 43 is a protecting group), and (vi) optionally, one or more protecting groups R 38 , R 39 , R 40 , R 41 , R 42 , and R 43 The method includes removing

[0369] In some embodiments, the protecting group is selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0370] The present disclosure also provides a method for synthesizing a polypeptide of SEQ ID NO: 16, comprising conjugating a compound of formula (IX-a) via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 18, and conjugating a compound of formula (IX-a) via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO: 19.

[0371] Formula (X) In another aspect, the disclosure provides a compound of formula (X), or a salt, solvate, or hydrate thereof:

[0372] [ka] In the formula, R 46 is H or a protecting group, and R 47 is H or a protecting group, and R 48 is H or a protecting group, and R 49 is H or a protecting group, and R 50 is H or a protecting group, and R 51 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0373] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups.

[0374] In some embodiments, R 46 is H or tert-butyl. In some embodiments, R 47 is H or tert-butyl. In some embodiments, R 48 is H or Boc. In some embodiments, R 49 is H or Fmoc. In some embodiments, R50 is H or tert-butyl. In some embodiments, R 51 is H or tert-butyl. In some embodiments, R 46 , R 47 , R 48 , R 49 , R 50 , and R 51 At least one of R 46 , R 47 , R 48 , R 49 , R 50 , and R 51 At least one of is a protecting group.

[0375] In some embodiments, formula (X) has formula (Xa):

[0376] [ka] The compound has the structure: Boc-Tyr(tBu)-Aib-Gln(trt)-Gly-OH.

[0377] In some embodiments, the compound of Formula (X) or (Xa) is a solvate.

[0378] In some embodiments, the solvate is formed from ethanol or isopropyl alcohol.

[0379] In some embodiments, the compound of Formula (X) or (Xa) is a desolvate.

[0380] In some embodiments, the compound of Formula (X) or (Xa) is crystalline.

[0381] In some embodiments, the compound of Formula (Xa) is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 18.1° and 18.7° ±0.2 degrees in combination with one or more peaks selected from 5.7°, 8.7°, 13.7°, 14.3°, 15.9°, and 16.2°.

[0382] In some embodiments, the compound of Formula (Xa) is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 5.9° and 10.5° ±0.2 degrees in combination with one or more peaks selected from 7.1°, 8.9°, 14.6°, and 16.6°.

[0383] In some embodiments, the compound of Formula (Xa) is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 7.8° and 20.3° ±0.2 degrees in combination with one or more peaks selected from 5.8°, 15.5°, and 19.5°.

[0384] In some embodiments, the compound of Formula (Xa) is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 5.9° and 7.4° ±0.2 degrees in combination with one or more peaks selected from 6.5°, 6.9°, and 14.8°.

[0385] The present disclosure also provides a method of synthesizing a compound of formula (X), comprising: (i) Formula (Y 保護 ) and compounds of formula (S 保護2 ) to react with a compound of formula (Y 保護 -S 保護2 forming a compound of

[0386] [ka] (In the formula, R 46 , R 47 , R 48 , and R 52 is a protecting group) (ii) Protecting group R 52 By removing (Y 保護 -S 保護1 forming a compound of

[0387] [ka] (iii) Equation (Y 保護 -S 保護1 ) and compounds of formula (K 保護2 ) to react with a compound of formula (Y 保護 -S 保護1 -K 保護2 forming a compound of

[0388] [ka] (In the formula, R 49 and R 53 is a protecting group) (iv) Protecting group R 53 By removing (Y 保護 -S 保護1 -K 保護1 forming a compound of

[0389] [ka] (v) Formula (Y 保護 -S 保護1 -K 保護1 ) and a compound of formula (Y 保護2 ) to form a compound of formula (X),

[0390] [ka] (In the formula, R 50 and R 51 is a protecting group), and (vi) optionally, one or more protecting groups R 46 , R 47 , R 48 , R 49 , R 50 , and R 51The method includes removing

[0391] In some embodiments, the protecting group is selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0392] The present disclosure also provides a method for synthesizing a polypeptide of SEQ ID NO: 16, comprising conjugating a compound of formula (Xa) via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 20, and conjugating a compound of formula (Xa) via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO: 21.

[0393] Formula (XI) In another aspect, the present disclosure provides a compound of formula (XI), or a salt, solvate, or hydrate thereof:

[0394] [ka] In the formula, R 64 is H or a protecting group, and R 65 is H or a protecting group, and R 66 is H or a protecting group, or a salt, solvate, or hydrate thereof. 64 and R 65 is a protecting group, and R 66 is H or a protecting group. In some embodiments, R 64 and R 65 is a protecting group, and R 66 is H.

[0395] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups.

[0396] In some embodiments, formula (XI) is Fmoc-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-OH having the structure of formula (XI-a):

[0397] [ka]

[0398] In some embodiments, the compound of formula (XI-a) is crystalline.

[0399] In some embodiments, the compound of formula (XI-a) is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 6.1° and 8.5° ±0.2 degrees in combination with one or more peaks selected from 5.8°, 16.9°, 18.5°, 18.8°, 19.3°, and 20.9°.

[0400] Formula (XII) In another aspect, the present disclosure provides a compound of formula (XII), or a salt, solvate, or hydrate thereof:

[0401] [ka] In the formula, R 61 is H or a protecting group, and R 62 is H or a protecting group, and R 63 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0402] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups. 61 is H or tert-butyl. In some embodiments, R 62 is H or tert-butyl. In some embodiments, R 63 is H or tert-butyl. In some embodiments, R 61 , R 62 , and R 63 At least one of R61 , R 62 , and R 63 At least one of is a protecting group.

[0403] In some embodiments, Formula (XII) has Formula (XII-a):

[0404] [ka] The side chain has the structure GGG.

[0405] In some embodiments, the compound of Formula (XII) or (XII-a) is a desolvate or anhydrous.

[0406] In some embodiments, the compound of Formula (XII) or (XII-a) is crystalline.

[0407] In some embodiments, the compound of Formula (XII-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 11.4° ±0.2 degrees in combination with one or more peaks selected from 6.0°, 8.9°, 12.7°, 13.6°, 14.6°, 17.0°, and 18.8°.

[0408] In some embodiments, the compound of Formula (XII-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 10.6° ±0.2 degrees in combination with one or more peaks selected from 7.1°, 12.1°, 13.6°, 14.2°, 15.2°, 16.0°, and 16.8°.

[0409] In some embodiments, the compound of Formula (XII-a) is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 10.1° and 15.5° ±0.2 degrees in combination with one or more peaks selected from 6.1°, 8.7°, 11.4°, 16.6°, and 19.2°.

[0410] Formula (XIII) In another aspect, the present disclosure provides a compound of formula (XIII), or a salt, solvate, or hydrate thereof:

[0411] [ka] In the formula, R 67 is H or a protecting group, and R 68 is H or a protecting group, and R 69 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0412] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups.

[0413] In some embodiments, R 67 is H or tert-butyl. In some embodiments, R 68 is H or tert-butyl. In some embodiments, R 69 is H or tert-butyl. In some embodiments, R 67 , R 68 , and R 69 At least one of R 67 , R 68 , and R 69 At least one of R is a protecting group. 67 and R 69 is H and R 68 is a protecting group.

[0414] In some embodiments, Formula (XIII) has Formula (XIII-a):

[0415] [ka] The amino acid sequence of the amino acid sequence of the present invention is H-Ala-Pro-Pro-Pro-Ser(tBu)-NH2 (HAPPPS-NH2), which has the structure:

[0416] In some embodiments, the compound of Formula (XIII) or (XIII-a) is a desolvate or anhydrous.

[0417] In some embodiments, the compound of Formula (XIII) or (XIII-a) is crystalline.

[0418] In some embodiments, the compound of Formula (XIII-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.0° ±0.2 degrees in combination with one or more peaks selected from 8.3, 9.7, and 11.2°.

[0419] In some embodiments, the compound of Formula (XIII-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.2° ±0.2 degrees in combination with one or more peaks selected from 5.3, 8.1, 14.4, and 16.2°.

[0420] Formula (XIV) In another aspect, the present disclosure provides a compound of formula (XIV), or a salt, solvate, or hydrate thereof:

[0421] [ka] In the formula, R 70 is H or a protecting group, and R 71 is H or a protecting group, and R 72 is H or a protecting group, and R 73 is H or a protecting group, and R 74 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0422] In some embodiments, each protecting group is independently selected from Boc, Fmoc, tert-butyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups.

[0423] In some embodiments, R 70 is H, Fmoc, or tert-butyl. In some embodiments, R 71 is H or tert-butyl. In some embodiments, R 72 is H or tert-butyl. In some embodiments, R 73 is H or tert-butyl. In some embodiments, R 74 is H, Fmoc, or tert-butyl. In some embodiments, R 70 , R 71 , R 72 , R 73 , and R 74 At least one of R 70 , R 71 , R 72 , R 73 , and R 74 At least one of R is a protecting group. 71 , R 72 , and R 73 is tert-butyl. In some embodiments, R 74 is H. In some embodiments, R 70 is Fmoc.

[0424] In some embodiments, formula (XIV) has formula (XIV-a):

[0425] [ka] The compound has the structure: Fmoc-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-NH2 (Fmoc-SSGAPPPS-NH2).

[0426] In some embodiments, the compound of formula (XIV) or (XIV-a) is a desolvate or anhydrous.

[0427] In some embodiments, the compound of formula (XIV) or (XIV-a) is crystalline.

[0428] In some embodiments, the compound of Formula (XIV-a) is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.5° ±0.2 degrees in combination with one or more peaks selected from 6.1, 8.7, 10.6, 15.0, 16.1, and 18.6°.

[0429] use In some embodiments, the polypeptide compounds described herein can be used to synthesize the active polypeptide GGG (SEQ ID NO: 12, 39 amino acids). In some embodiments, some of the compounds disclosed herein are conjugated with other polypeptide fragments to form full-length GGG. In some aspects, the crystalline compounds of the present invention (such as the Y-Aib-QG tetramer of formula (VI-a) and the Fmoc-GPSSG-NH2 pentamer of formula (XI-a)) can be used as pure, stable, and easy-to-use intermediates for the high-yield synthesis of GGG. In some embodiments, crystallization of the compounds can help eliminate impurities from the crude material. Compared to amorphous crude material, the crystalline compound can exhibit improved purity (e.g., as measured by UPLC-MS analysis). In some embodiments, crystallization of the crude compound reduces the required solvent flow, providing a "greener," more ecologically desirable process. In some embodiments, crystallization of a crude compound spiked with 1% dimer (impurity) can result in a crystalline compound with almost complete elimination of the dimer. In some embodiments, crystallization also improves physical properties, such as hygroscopicity. In some embodiments, crystalline compounds (such as the Y-Aib-QG tetramer of formula (VI-a) and the Fmoc-GPSSG-NH2 pentamer of formula (XI-a)) may exhibit significantly reduced weight gain due to moisture absorption compared to amorphous compounds.

[0430] In some embodiments, the polypeptide compounds described herein can be used to synthesize the active polypeptide OXM (SEQ ID NO: 16, 34 amino acids). In some embodiments, some of the compounds disclosed herein are conjugated with other polypeptide fragments to form full-length OXM. In some embodiments, the crystalline compounds of the present invention (e.g., H(dnp)-Aib-QG tetramer of formula (VII-a), H(trt)-Aib-QG tetramer of formula (VIII-a), DYSK tetramer of formula (IX-a), YSKY tetramer of formula (Xa), and Fmoc-GPSSG-NH2 pentamer of formula (XI-a)) can be used as pure, stable, and easy-to-use intermediates for the synthesis of OXM in high yields. In some embodiments, crystallization of the compounds can help eliminate impurities from the crude material. Compared to the amorphous crude material, the crystalline compounds can exhibit improved purity (e.g., as measured by UPLC-MS analysis). In some embodiments, crystallization of crude compounds reduces the required solvent flow, providing a "greener," more ecologically desirable process. In some embodiments, crystallization of crude compounds spiked with 1% dimer (impurity) can result in crystalline compounds with nearly complete elimination of the dimer. In some embodiments, crystallization also improves physical properties, such as hygroscopicity. In some embodiments, crystalline compounds (e.g., H(dnp)-Aib-QG tetramer of Formula (VII-a), H(trt)-Aib-QG tetramer of Formula (VIII-a), DYSK tetramer of Formula (IX-a), YSKY tetramer of Formula (Xa), and Fmoc-GPSSG-NH2 pentamer of Formula (XI-a)) can exhibit significantly reduced weight gain due to moisture absorption compared to amorphous compounds.

[0431] In some embodiments, compounds of Formula (I), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI), and (XII) comprise one or more protecting groups. In some embodiments, the one or more protecting groups are selected from Fmoc, Boc, tert-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups, or combinations thereof. In some embodiments, one or more protecting groups are Fmoc. In some embodiments, one or more protecting groups are Boc. In some embodiments, one or more protecting groups are tert-butyl. In some embodiments, one or more protecting groups are trityl. In some embodiments, one or more protecting groups are benzyl. In some embodiments, one or more protecting groups are carboxybenzyl. In some embodiments, one or more protecting groups are methyl. In some embodiments, one or more protecting groups are allyl. In some embodiments, one or more protecting groups are cyclohexyl. In some embodiments, compounds are protected with Fmoc, Boc, tert-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, and trityl groups. In some embodiments, compounds are protected with an Fmoc group. In some embodiments, compounds are protected with Fmoc and Boc groups. In some embodiments, compounds are protected with Fmoc and tert-butyl groups. In some embodiments, compounds are protected with Fmoc and trityl groups. In some embodiments, compounds are protected with Fmoc and benzyl groups. In some embodiments, compounds are protected with a Boc group. In some embodiments, compounds are protected with Boc and tert-butyl groups. In some embodiments, compounds are protected with Boc and trityl groups. In some embodiments, compounds are protected with Boc and benzyl groups. In some embodiments, compounds are protected with a tert-butyl group. In some embodiments, compounds are protected with tert-butyl and trityl groups. In some embodiments, compounds are protected with tert-butyl and benzyl groups. In some embodiments, compounds are protected with a trityl group.In some embodiments, the compound is protected with trityl and benzyl groups. In some embodiments, the compound is protected with benzyl groups. In some embodiments, the compound is protected with Fmoc, Boc, and tert-butyl groups. In some embodiments, the compound is protected with Fmoc, Boc, and trityl groups. In some embodiments, the compound is protected with Fmoc, Boc, and benzyl groups. In some embodiments, the compound is protected with Fmoc, tert-butyl, and trityl groups. In some embodiments, the compound is protected with Fmoc, tert-butyl, and benzyl groups. In some embodiments, the compound is protected with Fmoc, trityl, and benzyl groups. In some embodiments, the compound is protected with Boc, tert-butyl, and trityl groups. In some embodiments, the compound is protected with Boc, tert-butyl, and benzyl groups. In some embodiments, the compound is protected with Fmoc, tert-butyl, and trityl groups.

[0432] As used herein, the following abbreviations have the meanings set forth herein: "API" means active pharmaceutical ingredient, "DCM" means dichloromethane, "DIC" means diisopropylcarbodiimide, "Oxyma" means ethyl cyanohydroxyiminoacetate, "DTT" means dithiothreitol, "Fmoc" means fluorenylmethyloxycarbonyl chloride, "GGG" means retatortide, "IPA" means isopropyl alcohol, and "MTBE" means methyl tert-butyl ether. where "OXM" means mazdutide, "Pip" means piperidine, "PyBOP" means (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate), "SPPS" means solid phase peptide synthesis, "TFA" means trifluoroacetic acid, "TNTU" means 2-(5-norbornene-2,3-dicarboximido)-1,1,3,3-tetramethyluronium tetrafluoroborate, "TZP" means tirzepatide, and "UPLC" means ultra performance liquid chromatography.

[0433] In some embodiments, single-letter amino acid abbreviations are shown in bold, with atoms shown as non-bold text to distinguish them from the single-letter amino acid abbreviations. In some embodiments, amino acids are represented by their three-letter abbreviations. As used herein, when an amino acid abbreviation appears with a number above the amino acid, that number refers to the position of the corresponding amino acid in the final product. Numbers are provided for convenience, and the appearance or non-appearance of such numbers in a sequence does not affect the amino acid sequence or peptide depicted in such sequence.

[0434] As used herein, the term "protected" means that a protecting group is attached at the indicated position.

[0435] As used herein, the term "crystalline" includes all crystalline forms that have a distinguishable X-ray pattern.

[0436] As used herein, "amino acid" refers to natural and unnatural synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function similarly to the naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code. Amino acids may be referred to herein by either their commonly known three letter symbols or the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Amino acids include side chains and polypeptide backbone moieties.

[0437] A "peptide" or "polypeptide" is a linked sequence of two or more amino acids linked by peptide bonds. Polypeptides can be natural, synthetic, or a modified or combination of natural and synthetic.

[0438] As used herein, the term "protecting group" or "amino acid protecting group" refers to a group that protects the acid or amine moiety of an amino acid or a reactive moiety on the side chain of an amino acid. An "acid moiety" includes, for example, a carboxylic acid group (-COOH). An "amine moiety" includes, for example, a primary amine group (-NH), a secondary amine group (-NH-), an amide group (-C(O)-NH), and a guanidinium group ([-NHC(NH)-NH]). + ). The acid or amine moiety can be part of the terminal amino acid in a peptide or polypeptide, or on the side chain of a non-terminal amino acid in a peptide or polypeptide. Other reactive moieties on the side chain of amino acids include, for example, hydroxy (-OH) and thiol (-SH) groups.

[0439] The protecting group may be a removable group known in the art to (i) protect a reactive group (such as an amine or carboxylic acid group) against undesired reactions during synthetic procedures, e.g., to block or protect the functionality of a reactive group during reactions involving other functional portions of a compound, and (ii) to be selectively deprotected in a multiply protected structure without affecting other protecting groups. Suitable protecting groups and methods for introducing and removing such groups include those known in the art, such as those described in T.W. Green and P.G.M. Buts, Greene's Protective Groups in Organic Synthesis, John Wiley and Sons, 2007, and Isidro-Llobet et al., Amino Acid-Protecting Groups, Chem. Rev., 2009, 109(6), 2455-2504, the entire contents of which are incorporated herein.

[0440] Suitable protecting groups for aspartic acid (Asp) include, but are not limited to, tert-butyl (t-Bu), 3-methyl-3-pentyl (mpe), allyl, and 4-{N-[1-(4,4-dimethyl-2,6-dioxocyclohexylidene)-3-methylbutyl]amino}benzyl (DMAB). In some embodiments, the protecting group for aspartic acid (Asp) is t-Bu or mpe.

[0441] Suitable protecting groups for serine (Ser), threonine (Thr), or tyrosine (Tyr) include, but are not limited to, t-Bu and triphenylmethyl (trityl or trt). In some embodiments, the protecting group for serine (Ser), threonine (Thr), or tyrosine (Tyr) is t-Bu or TRT.

[0442] Suitable protecting groups for glutamic acid (Glu) include, but are not limited to, t-Bu, trt, allyl, and DMAB. In some embodiments, the protecting group for glutamic acid (Glu) is t-Bu or trt.

[0443] Suitable protecting groups for glutamine (Gln) include, but are not limited to, trt, 4-methoxytrityl (4-methyltrityl, or MTT), acetamidomethyl (ACM), and trimethoxybenzyl (TMOB). In some embodiments, the protecting group for glutamine (Gln) is TRT.

[0444] Suitable protecting groups for lysine (Lys) include, but are not limited to, t-butoxycarbonyl (Boc), allyloxycarbonyl (Alloc), 4-phenylacetoxybenzyloxycarbonyl (PhAc), MTT, 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl (ivDde), and 2-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl (Dde). In some embodiments, the protecting group for lysine (Lys) is Boc, MTT, or Alloc.

[0445] Suitable protecting groups for tryptophan (Trp) include, but are not limited to, Boc and formyl. In some embodiments, the protecting group for tryptophan (Trp) is Boc.

[0446] Suitable protecting groups for histidine (His) include, but are not limited to, Boc, trt, and 2,4-dinitrophenyl (dnp). In some embodiments, the protecting group for histidine (His) is Boc, trt, or dnp.

[0447] Exemplary acid protecting groups include esters, such as substituted and unsubstituted C1-C8 lower alkyl (e.g., methyl, ethyl, t-butyl), methoxymethyl, methylthiomethyl, 2,2,2-trichloroethyl, tetrahydropyranyl, substituted and unsubstituted phenylalkyl (e.g., benzyl), and substituted derivatives thereof (e.g., alkoxybenzyl, nitrobenzyl), cinnamyl, dialkylaminoalkyl (e.g., dimethylaminoethyl), trimethylsilyl, substituted and unsubstituted amides and hydrazides (e.g., amides and hydrazides of N,N-dimethylamine), 7-nitroindole, hydrazine, N-phenylhydrazine, acyloxyalkyl (e.g., pivaloyloxymethyl, propionyloxymethyl), aroyloxyalkyl (e.g., benzoyloxyethyl), alkoxycarbonylalkyl (e.g., methoxycarbonylmethyl), cyclohexyloxycarbonylmethyl, Examples of the alkyl group include alkoxycarbonyloxyalkyl (e.g., t-butyloxycarbonyloxymethyl), alkoxycarbonylaminoalkyl (e.g., t-butyloxycarbonylaminomethyl), alkylaminocarbonylaminoalkyl (e.g., methylaminocarbonylaminomethyl), acylaminoalkyl (e.g., acetylaminomethyl), heterocyclylcarbonyloxyalkyl (e.g., 4-methylpiperazinyl-carbonyloxymethyl), dialkylaminocarbonylalkyl (e.g., dimethylaminocarbonyl-methyl), (5-(lower alkyl)-2-oxo-1,3-dioxolen-4-yl)alkyl (e.g., (5-t-butyl-2-oxo-1,3-dioxolen-4-yl)methyl), and (5-phenyl-2-oxo-1,3-dioxolen-4-yl)alkyl (e.g., (5-phenyl-2-oxo-1,3-dioxolen-4-yl)methyl).

[0448] Exemplary amine and / or amide protecting groups include, but are not limited to, acyl (e.g., formyl, acetyl, chloroacetyl, trichloroacetyl, o-nitrophenylacetyl, o-nitrophenoxy-acetyl, trifluoroacetyl, acetoacetyl, 4-chlorobutyryl, isobutyryl, o-nitrocinnamoyl, picolinoyl, acylisothiocyanate, aminocaproyl, benzoyl), acyloxy (e.g., methoxy-carbonyl, 9-fluorenylmethoxycarbonyl, 2,2,2-trifluoroethoxycarbonyl, 2-trimethylsilylethoxy-carbonyl, vinyloxycarbonyl, allyloxycarbonyl, t-butyloxycarbonyl (Boc), 1,1-dimethyl-propynyloxycarbonyl, benzyloxycarbonyl (Cbz), p-nitrobenzyloxycarbonyl, 2,4-dichloro-benzyloxycarbonyl), 9-xanthenyl, and trityl. Further exemplary amide protecting groups include, but are not limited to, o-nitrocinnamoyl, picolinoyl, aminocaproyl, benzoyl, acyloxy (e.g., methoxycarbonyl, 9-fluorenylmethoxycarbonyl, 2,2,2-trifluoroethoxycarbonyl, 2-trimethylsilylethoxycarbonyl, vinyloxycarbonyl, allyloxycarbonyl, t-butyloxycarbonyl (Boc), 1,1-dimethyl-propynyloxycarbonyl, benzyloxycarbonyl (Cbz), p-nitrobenzyloxycarbonyl, and 2,4-dichloro-benzyloxycarbonyl). Exemplary indole protecting groups include, but are not limited to, formyl (For) and t-butyloxycarbonyl (Boc). Exemplary imidazole protecting groups include, but are not limited to, tosyl (To), benzyloxymethyl (Bom), trityl (Trt), and t-butyloxycarbonyl (Boc). Exemplary guanidinium protecting groups include, but are not limited to, 2,2,4,6,7-pentamethyl-2,3-dihydrobenzofuran-5-sulfonyl (Pbf) and t-butyloxycarbonyl (Boc).

[0449] Exemplary hydroxyl protecting groups include, but are not limited to, unsubstituted or substituted alkyl (e.g., t-butyl, allyl, benzyl, methoxymethyl, tetrahydropyranyl, o-nitrobenzyl), silyl (e.g., t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (TBDPS)), acyl (e.g., acetyl, benzoyl, pivaloyl). Exemplary thiol protecting groups include, but are not limited to, p-methylbenzyl (Meb), acetamidomethyl (Acm), and trityl (Trt).

[0450] Enumerated Embodiments Embodiment 1. A compound of formula (I), or a salt, solvate, or hydrate thereof,

[0451] [ka] In the formula, R 1 is H or a protecting group, and R 2 is H or a protecting group, and R 3 is H or a protecting group, and R 4 is H or a protecting group, and R 1 , R 2 , R 3 , and R 4 or a salt, solvate, or hydrate thereof, wherein at least one of:

[0452] Embodiment 2.R 1 is H, Fmoc, or Boc.

[0453] Embodiment 3.R 2 The compound of any one of embodiments 1-2, wherein is H or t-butyl.

[0454] Embodiment 4.R 4The compound of any one of embodiments 1-3, wherein is H or benzyl.

[0455] Embodiment 5.R 3 The compound of any one of embodiments 1-4, wherein is H, t-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, or trityl.

[0456] Embodiment 6.R 1 , R 2 , R 3 and R 4 The compound of any one of embodiments 1-5, wherein at least two of are H.

[0457] Embodiment 7.R 1 , R 2 , R 3 , and R 4 The compound of any one of embodiments 1-6, wherein at least two of are protecting groups.

[0458] Embodiment 8. The compound has the formula:

[0459] [ka] 2. The compound of embodiment 1, wherein

[0460] Embodiment 9. The compound has the formula:

[0461] [ka] 2. The compound of embodiment 1, wherein

[0462] Embodiment 10. The compound has the formula:

[0463] [ka] 2. The compound of embodiment 1, wherein

[0464] Embodiment 11. The compound has the formula:

[0465] [ka] 2. The compound of embodiment 1, wherein

[0466] Embodiment 12. The compound has the formula:

[0467] [ka] 2. The compound of embodiment 1, wherein

[0468] Embodiment 13. The compound of any one of embodiments 1 to 12, wherein the compound is a solvate.

[0469] Embodiment 14. The compound of embodiment 13, wherein the solvate is formed from methyl t-butyl ether (MTBE) or a mixture containing MTBE.

[0470] Embodiment 15. The compound of any one of embodiments 1 to 12, wherein the compound is crystalline.

[0471] Embodiment 16. The compound of embodiment 15, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.2° ± 0.2 degrees, in combination with one or more peaks selected from 8.4°, 8.8°, 10.4°, 15.5°, 17.1°, and 17.7°.

[0472] Embodiment 17. The compound of embodiment 15, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 6.1° ± 0.2 degrees in combination with one or more peaks selected from 10.3°, 14.9°, 16.8°, 18.1°, and 18.2°.

[0473] Embodiment 18. The compound of embodiment 15, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 6.2° ± 0.2 degrees, in combination with one or more peaks selected from 8.9°, 12.3°, 14.9°, 15.4°, and 21.8°.

[0474] Embodiment 19. A method of synthesizing a compound according to embodiment 1, comprising: -Formula (Y 保護 )

[0475] [ka] In the formula, R 1 and R 2 is a protecting group, 保護 ) and a compound of formula (Aib 保護 )

[0476] [ka] In the formula, R 5 is a protecting group, 保護 ) to react with a compound of formula (Y 保護 -Aib 保護 forming a compound of

[0477] [ka] -R 5 and removing the protecting group of the formula (Y 保護 -Aib),

[0478] [ka] -Formula (Y 保護 -Aib) and a compound of formula (E 保護 )

[0479] [ka] In the formula, R 3 is a protecting group, 保護 ) to react with a compound of formula (Y 保護 -Aib-E 保護 forming a compound of

[0480] [ka] -Formula (Y 保護 -Aib-E 保護 ) and a compound of formula (G 保護 )

[0481] [ka] In the formula, R 4 is a protecting group, 保護 ) to react with a compound of formula (Y 保護 -Aib-E 保護 -G 保護 forming a compound of

[0482] [ka] and, optionally, -Protecting group R 1 , R 2 , R 3 , and R 4 removing one or more of the

[0483] Embodiment 20. A method for synthesizing a polypeptide of SEQ ID NO: 1, comprising conjugating a compound according to any one of embodiments 1 to 12 to the N-terminus of a polypeptide of SEQ ID NO: 2 via the C-terminus of the compound.

[0484] Embodiment 21. A method for synthesizing a polypeptide of SEQ ID NO: 26, comprising conjugating a compound of any one of embodiments 1 to 12 via the C-terminus of the compound to the N-terminus of a polypeptide of SEQ ID NO: 27.

[0485] Embodiment 22. A compound of formula (II) or a salt, solvate, or hydrate thereof,

[0486] [ka] In the formula, R 6 is H or a protecting group, and R 7 is H or a protecting group, and R 8 is H or a protecting group, and R 9 is H or a protecting group, and R 10 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0487] Embodiment 23. The compound of embodiment 22, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0488] Embodiment 24.R 6 The compound of any one of embodiments 22-23, wherein is H, Fmoc, or Boc.

[0489] Embodiment 25.R 7 The compound of any one of embodiments 22-24, wherein is H or t-butyl.

[0490] Embodiment 26.R 8 The compound of any one of embodiments 22-25, wherein is H or t-butyl.

[0491] Embodiment 27.R 9 The compound of any one of embodiments 22-26, wherein is H or t-butyl.

[0492] Embodiment 28.R 10 The compound of any one of embodiments 22-27, wherein is H or benzyl.

[0493] Embodiment 29.R 6 , R 7 , R8 , R 9 , and R 10 The compound of any one of embodiments 22-28, wherein at least one of is a protecting group.

[0494] Embodiment 30.R 6 , R 7 , R 8 , R 9 , and R 10 is H. The compound of any one of embodiments 22-29, wherein

[0495] Embodiment 31. The compound has the formula:

[0496] [ka] 23. The compound of embodiment 22, wherein

[0497] Embodiment 32. The compound of any one of embodiments 22-31, wherein the compound is a solvate.

[0498] Embodiment 33. The compound of embodiment 32, wherein the solvate is formed from a solvent comprising heptane.

[0499] Embodiment 34. The compound of any one of embodiments 22-31, wherein the compound is crystalline.

[0500] Embodiment 35. The compound of embodiment 34, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 7.7 to 7.9° ± 0.2 degrees, in combination with one or more peaks selected from 5.8°, 10.0°, 10.8-10.9°, 11.3-11.4°, 12.0-12.1°, 12.8°, 14.2-14.4°, and 16.8-17.0°.

[0501] Embodiment 36. The compound of embodiment 34, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 7.3° ± 0.2 degrees, in combination with one or more peaks selected from 5.1°, 5.7°, 7.6°, 9.5°, and 12.4°.

[0502] Embodiment 37. The compound of embodiment 34, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 7.8° ± 0.2 degrees in combination with one or more peaks selected from 8.5°, 11.5°, 12.0°, 12.8°, 14.3°, 15.5°, 20.2°, and 23.3°.

[0503] Embodiment 38. The compound of embodiment 34, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 8.2 to 8.3° ±0.2 degrees, in combination with one or more peaks selected from 5.9°, 7.7°, 9.2°, 10.2°, 11.3°, 13.8 to 13.9°, 15.5 to 15.7°, 17.1°, and 18.5°.

[0504] Embodiment 39. A method for synthesizing a polypeptide of SEQ ID NO: 1, comprising: (i) conjugating a compound described in any one of embodiments 22 to 31 via the N-terminus of the compound to the C-terminus of a polypeptide of SEQ ID NO: 3 to form a polypeptide of SEQ ID NO: 4; and (ii) conjugating the polypeptide of SEQ ID NO: 4 via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 5.

[0505] Embodiment 40. A compound of formula (III), or a salt, solvate, or hydrate thereof,

[0506] [ka] In the formula, R 11 is H or a protecting group, and R 12 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0507] Embodiment 41. The compound of embodiment 40, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0508] Embodiment 42.R 11 The compound of any one of embodiments 40-41, wherein is H or t-butyl.

[0509] Embodiment 43.R 12 The compound of any one of embodiments 40-42, wherein is H or t-butyl.

[0510] Embodiment 44.R 11 and R 12 The compound of any one of embodiments 40-43, wherein at least one of is a protecting group.

[0511] Embodiment 45.R 11 and R 12 The compound of any one of embodiments 40-43, wherein is H.

[0512] Embodiment 46. The compound has the formula:

[0513] [ka] 41. The compound of embodiment 40, wherein

[0514] Embodiment 47. The compound of any one of embodiments 40-46, wherein the compound is a solvate.

[0515] Embodiment 48. The compound of any one of embodiments 40-46, wherein the compound is crystalline.

[0516] Embodiment 49. The compound of embodiment 48, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 8.0° ± 0.2 degrees in combination with one or more peaks selected from 7.0°, 10.3°, 14.1°, 15.2°, 16.7°, 18.0°, 19.0°, 19.7°, 20.8°, and 21.9°.

[0517] Embodiment 50. The compound of embodiment 48, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 9.0° ± 0.2 degrees in combination with one or more peaks selected from 5.7°, 9.9°, 16.2°, 17.1°, 17.9°, 18.1°, 18.4°, 18.8°, 19.9°, 20.1°, and 22.5°.

[0518] Embodiment 51. The compound of embodiment 48, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 10.5° ± 0.2 degrees in combination with one or more peaks selected from 5.6°, 11.9°, 13.3°, 15.4°, 15.6°, 18.1°, 19.9°, and 21.1°.

[0519] Embodiment 52. The compound of embodiment 48, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 10.7° ± 0.2 degrees, in combination with one or more peaks selected from 4.9°, 14.8°, 20.3°, and 21.5°.

[0520] Embodiment 53. The compound of embodiment 48, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 10.5° ± 0.2 degrees in combination with one or more peaks selected from 5.9°, 10.5°, 10.9°, 12.1°, 13.1°, 15.9°, 17.5°, 20.9°, 21.1°, and 21.9°.

[0521] Embodiment 54. The compound of embodiment 48, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 7.8° ± 0.2 degrees in combination with one or more peaks selected from 11.3°, 11.5°, 15.4°, 15.6°, and 21.5°.

[0522] Embodiment 55. The compound of embodiment 48, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 10.0° ± 0.2 degrees in combination with one or more peaks selected from 8.1°, 12.5°, 13.5°, 14.7°, 17.8°, 18.8°, 20.0°, and 22.4°.

[0523] Embodiment 56. The compound of embodiment 48, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 21.1° ± 0.2 degrees in combination with one or more peaks selected from 5.6°, 10.5°, 10.8°, 11.9°, 15.4°, and 23.8°.

[0524] Embodiment 57. A method for synthesizing a polypeptide of SEQ ID NO: 1, comprising: (i) conjugating a compound described in any one of embodiments 40 to 46 via the N-terminus of the compound to the C-terminus of a polypeptide of SEQ ID NO: 6 to form a polypeptide of SEQ ID NO: 7; and (ii) conjugating the polypeptide of SEQ ID NO: 7 via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 8.

[0525] Embodiment 58. A compound of formula (IV) or a salt, solvate, or hydrate thereof,

[0526] [ka] In the formula, R 13 is H or a protecting group, and R 13* is H or a protecting group, and R 14 is H or a protecting group, and R 15is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0527] Embodiment 59. The compound of embodiment 58, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0528] Embodiment 60.R 13 The compound of any one of embodiments 58-59, wherein is H, Fmoc, or Boc.

[0529] Embodiment 61.R 13* The compound of any one of embodiments 58-60, wherein is H.

[0530] Embodiment 62.R 14 The compound of any one of embodiments 58-61, wherein is H or t-butyl.

[0531] Embodiment 63.R 15 The compound of any one of embodiments 58-62, wherein is H or t-butyl.

[0532] Embodiment 64.R 13 , R 13* , R 14 , and R 15 The compound of any one of embodiments 58-63, wherein at least one of is a protecting group.

[0533] Embodiment 65.R 13 , R 13* , R 14 , and R 15 is H. The compound of any one of embodiments 58-63, wherein

[0534] Embodiment 66. The compound has the formula:

[0535] [ka] 59. The compound of embodiment 58, wherein

[0536] Embodiment 67. The compound of any one of embodiments 58-66, wherein the compound is a solvate.

[0537] Embodiment 68. The compound of any one of embodiments 58-66, wherein the compound is crystalline.

[0538] Embodiment 69. The compound of embodiment 68, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.1° ± 0.2 degrees in combination with one or more peaks selected from 4.3°, 6.1°, 8.0°, 10.1°, and 18.7°.

[0539] Embodiment 70. The compound of embodiment 68, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.2° ± 0.2 degrees in combination with one or more peaks selected from 6.0°, 6.7°, 10.0°, 10.3°, 16.4°, 17.8°, 18.3°, 19.4°, and 22.4°.

[0540] Embodiment 71. A method for synthesizing a polypeptide of SEQ ID NO: 1, comprising: (i) conjugating a compound described in any one of claims 58 to 66 via the N-terminus of the compound to the C-terminus of a polypeptide of SEQ ID NO: 9 to form a polypeptide of SEQ ID NO: 7; and (ii) conjugating the polypeptide of SEQ ID NO: 7 via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 8.

[0541] Embodiment 72. The compound has the formula

[0542] [ka] is a compound of In the formula, R 54 is H or a protecting group, and R 55 is H or a protecting group, and R 56 is H or a protecting group.

[0543] Embodiment 73. The compound of embodiment 72, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0544] Embodiment 74.R 54 The compound of any one of embodiments 72-73, wherein is H or tert-butyl.

[0545] Embodiment 75.R 55 The compound of any one of embodiments 72-74, wherein is H or tert-butyl.

[0546] Embodiment 76.R 56 The compound of any one of embodiments 72-75, wherein is H or Boc.

[0547] Embodiment 77.R 54 , R 55 , and R 56 The compound of any one of embodiments 72-76, wherein at least one of: is H.

[0548] Embodiment 78.R 54 , R 55 , and R 56 The compound of any one of embodiments 72-77, wherein at least one of is a protecting group.

[0549] Embodiment 79. The compound has the formula:

[0550] [ka] 73. The compound of embodiment 72, wherein

[0551] Embodiment 80. The compound of any one of embodiments 72-79, wherein the compound is a solvate.

[0552] Embodiment 81 The compound of embodiment 80, wherein the compound is a solvate formed from acetone.

[0553] Embodiment 82. The compound of any one of embodiments 72 to 79, wherein the compound is a desolvate.

[0554] Embodiment 83. The compound of any one of embodiments 72-79, wherein the compound is crystalline.

[0555] Embodiment 84. The compound of embodiment 83, wherein the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 5.8° and 18.5° ±0.2 degrees in combination with one or more peaks selected from 8.6°, 9.4°, 12.9°, 13.8°, 17.2°, and 19.4°.

[0556] Embodiment 85. The compound of embodiment 83, wherein the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta ±0.2 degrees of 7.0-7.1° and 7.5-7.7° in combination with one or more peaks selected from 5.3-5.4°, 9.7-9.9°, and 14.7-14.9.

[0557] Embodiment 86. The compound of embodiment 83, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 8.3° ±0.2 degrees in combination with one or more peaks selected from 6.3°, 11.4°, 14.3°, and 16.6°.

[0558] Embodiment 87. The compound of embodiment 83, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.2° ±0.2 degrees, in combination with one or more peaks selected from 6.8°, 8.6°, 15.8°, and 18.9°.

[0559] Embodiment 88. The compound of embodiment 83, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 6.1° ±0.2 degrees in combination with one or more peaks selected from 6.3°, 7.8°, 10.0°, and 12.4°.

[0560] Embodiment 89. A method of synthesizing a compound of embodiment 72, comprising: - a compound of formula (Fmoc-G) and a compound of formula (P 保護 ) to react with a compound of the formula (Fmoc-GP 保護 forming a compound of

[0561] [ka] (In the formula, R 57 is a protecting group) -Protecting group R 57 to form the compound (Fmoc-GP),

[0562] [ka] - a compound of formula (Fmoc-GP) and a compound of formula (S 保護2 ) to form a compound of the formula (Fmoc-GPS 保護2 forming a compound of

[0563] [ka] (In the formula, R 54 and R 58 is a protecting group) -Protecting group R 58 (Fmoc-GPS 保護1 forming a compound of

[0564] [ka] -Formula(Fmoc-GPS 保護1 ) and compounds of formula (S 保護2) to form a compound of the formula (Fmoc-GPS 保護1 -S 保護2 forming a compound of

[0565] [ka] (In the formula, R 55 and R 59 is a protecting group) -Protecting group R 59 (Fmoc-FG 保護1 -S 保護1 -S 保護1 forming a compound of

[0566] [ka] -Formula(Fmoc-GPS 保護1 -S 保護1 ) and compounds of formula (G 保護 ) to form a compound of the formula (Fmoc-GPS 保護1 -S 保護1 -G 保護 )

[0567] [ka] forming a compound of (In the formula, R 60 is a protecting group), -Formula(Fmoc-GPS 保護1 -S 保護1 -G 保護 37. A compound of formula (I) above,

[0568] [ka] and optionally one or more protecting groups R 54 , R 55 , and R 56 The method of claim 1, further comprising:

[0569] Embodiment 90. The method of embodiment 89, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0570] Embodiment 91. A method for synthesizing a polypeptide of SEQ ID NO: 12, comprising: (i) conjugating a compound described in any one of embodiments 72 to 79 via the C-terminus of the compound to the N-terminus of a polypeptide of SEQ ID NO: 22; and (ii) conjugating the resulting compound via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO: 23.

[0571] Embodiment 92. A method for synthesizing a polypeptide of SEQ ID NO: 24, comprising conjugating a compound according to any one of embodiments 72 to 79 to the N-terminus of a polypeptide of SEQ ID NO: 22 via the C-terminus of the compound.

[0572] Embodiment 93. A method for synthesizing a polypeptide of SEQ ID NO: 16, comprising conjugating a compound according to any one of embodiments 72 to 79 to the C-terminus of a polypeptide of SEQ ID NO: 25 via the N-terminus of the compound.

[0573] Embodiment 94. A method for synthesizing a polypeptide of SEQ ID NO: 1, comprising: (i) conjugating a compound described in any one of embodiments 72 to 79 via the C-terminus of the compound to the N-terminus of a polypeptide of SEQ ID NO: 8; and (ii) conjugating the resulting compound via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO: 9.

[0574] Embodiment 95. A compound of formula (V), or a salt, solvate, or hydrate thereof,

[0575] [ka] In the formula, R 16 is H or a protecting group, and R 17 is H or a protecting group, and R 18is H or a protecting group, and R 19 is H or a protecting group, and R 20 is H or a protecting group, and R 21 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0576] Embodiment 96. The compound of embodiment 95, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0577] Embodiment 97.R 16 The compound of any one of embodiments 95-96, wherein is H, Fmoc, or Boc.

[0578] Embodiment 98.R 17 The compound of any one of embodiments 95-97, wherein is H or t-butyl.

[0579] Embodiment 99.R 18 The compound of any one of embodiments 95-98, wherein is H or t-butyl.

[0580] Embodiment 100.R 19 The compound of any one of embodiments 95-99, wherein is H or t-butyl.

[0581] Embodiment 101.R 20 The compound of any one of embodiments 95-100, wherein is H or t-butyl.

[0582] Embodiment 102.R 21 The compound according to any one of embodiments 95-101, wherein is H or benzyl.

[0583] Embodiment 103.R 16 , R 17 , R 18 , R 19 , R 20 , and R 21 The compound of any one of embodiments 95-102, wherein at least one of is a protecting group.

[0584] Embodiment 104.R 16 , R 17 , R 18 , R 19 , R 20 , and R 21 The compound of any one of embodiments 95-102, wherein is H.

[0585] Embodiment 105. The compound has the formula:

[0586] [ka] 96. The compound of embodiment 95, wherein

[0587] Embodiment 106. The compound of any one of embodiments 95-105, wherein the compound is a solvate.

[0588] Embodiment 107. The compound of any one of embodiments 95-105, wherein the compound is crystalline.

[0589] Embodiment 108. The compound of embodiment 107, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.8 to 6.1° ± 0.2 degrees, in combination with one or more peaks selected from 6.7 to 7.1° and 8.8 to 9.0°.

[0590] Embodiment 109. The compound of embodiment 107, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.0 to 5.2° ± 0.2 degrees in combination with one or more peaks selected from 5.3 to 5.4°, 5.7 to 6.0°, 6.1 to 6.2°, 7.6 to 7.9°, and 8.7 to 9.1°.

[0591] Embodiment 110. A method for synthesizing a polypeptide of SEQ ID NO: 1, comprising: (i) conjugating a compound described in any one of embodiments 95 to 105 via the N-terminus of the compound to the C-terminus of a polypeptide of SEQ ID NO: 3 to form a polypeptide of SEQ ID NO: 10; and (ii) conjugating the polypeptide of SEQ ID NO: 10 via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 11.

[0592] Embodiment 111. A compound of formula (VI) or a salt, solvate, or hydrate thereof,

[0593] [ka] In the formula, R 22 is H or a protecting group, and R 23 is H or a protecting group, and R 24 is H or a protecting group, and R 25 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0594] Embodiment 112. The compound of embodiment 111, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0595] Embodiment 113.R 22 The compound of any one of embodiments 111-112, wherein is H or Boc.

[0596] Embodiment 114.R 23 The compound according to any one of embodiments 111 to 113, wherein is H or tert-butyl.

[0597] Embodiment 115.R 24 The compound according to any one of embodiments 111-114, wherein is H or trityl.

[0598] Embodiment 116.R 25The compound according to any one of embodiments 111 to 115, wherein is H or tert-butyl.

[0599] Embodiment 117.R 22 , R 23 , R 24 , and R 25 The compound of any one of embodiments 111-116, wherein at least one of: is H.

[0600] Embodiment 118.R 22 , R 23 , R 24 , and R 25 The compound of any one of embodiments 111-117, wherein at least one of is a protecting group.

[0601] Embodiment 119. The compound has the formula (VI-a):

[0602] [ka] 112. The compound of embodiment 111, wherein

[0603] Embodiment 120. The compound of any one of embodiments 111 to 119, wherein the compound is a solvate.

[0604] Embodiment 121. The compound of embodiment 120, wherein the compound is a solvate formed from pentyl acetate, a mixture comprising pentyl acetate, ethyl acetate, or a mixture comprising 2-methyltetrahydrofuran and t-amyl methyl ether.

[0605] Embodiment 122. The compound according to embodiment 121, wherein the mixture comprising pentyl acetate is selected from a mixture comprising pentyl acetate and t-butyl ethyl ether, a mixture comprising pentyl acetate and t-amyl methyl ether, or a mixture comprising pentyl acetate and heptane.

[0606] Embodiment 123. The compound of any one of embodiments 111 to 119, wherein the compound is a desolvate or a partial desolvate.

[0607] Embodiment 124. The compound of any one of embodiments 111 to 119, wherein the compound is crystalline.

[0608] Embodiment 125. The compound of embodiment 124, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 6.3 to 6.4° ±0.2 degrees in combination with one or more peaks selected from 4.5°, 7.1°, 13.0 to 13.1°, 15.9 to 16.0°, and 18.4 to 18.6°.

[0609] Embodiment 126. The compound of embodiment 124, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.0 to 7.2° ±0.2 degrees in combination with one or more peaks selected from 5.0 to 5.4°, 7.6 to 7.7°, 8.8 to 8.9°, 9.4 to 9.5°, and 12.5 to 12.7°.

[0610] Embodiment 127. A method for synthesizing a compound according to embodiment 111, comprising: -Formula (Y 保護 ) and compounds of formula (Aib 保護 ) to react with a compound of formula (Y 保護 -Aib 保護 forming a compound of

[0611] [ka] (In the formula, R 22 , R 23 , and R 26 is a protecting group) -Protecting group R 26 By removing (Y 保護 -Aib),

[0612] [ka] -Formula (Y 保護 -Aib) and the compound of formula (Q 保護2 ) to react with a compound of formula (Y 保護 -Aib-Q 保護2 forming a compound of

[0613] [ka] (In the formula, R 24 and R 27 is a protecting group) -Protecting group R 27 By removing (Y 保護 -Aib-Q 保護1 forming a compound of

[0614] [ka] -Formula (Y 保護 -Aib-Q 保護1 ) and compounds of formula (G 保護 ) to form a compound of formula (VI),

[0615] [ka] (In the formula, R 25 is a protecting group), and optionally one or more protecting groups R 22 , R 23 , R 24 , and R 25 The method of claim 1, further comprising:

[0616] Embodiment 128. The method of embodiment 127, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0617] Embodiment 129. A method for synthesizing a polypeptide of SEQ ID NO: 12, comprising conjugating a compound according to any one of embodiments 111 to 119 to the N-terminus of a polypeptide of SEQ ID NO: 13 via the C-terminus of the compound.

[0618] Embodiment 130. A method for synthesizing a polypeptide of SEQ ID NO: 14, comprising conjugating a compound according to any one of embodiments 111 to 119 to the N-terminus of a polypeptide of SEQ ID NO: 15 via the C-terminus of the compound.

[0619] Embodiment 131. A compound of formula (VII) or a salt, solvate, or hydrate thereof, wherein

[0620] [ka] In the formula, R 28 is H or a protecting group, and R 29 is H or a protecting group, and R 30 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0621] Embodiment 132. The compound of embodiment 131, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0622] Embodiment 133.R 28 The compound according to any one of embodiments 131-132, wherein is H or Boc.

[0623] Embodiment 134.R 29 The compound according to any one of embodiments 131-133, wherein is H or trityl.

[0624] Embodiment 135.R 30 The compound according to any one of embodiments 131-134, wherein is H or tert-butyl.

[0625] Embodiment 136.R28 , R 29 , and R 30 The compound of any one of embodiments 131-135, wherein at least one of: is H.

[0626] Embodiment 137.R 28 , R 29 , and R 30 The compound of any one of embodiments 131-136, wherein at least one of is a protecting group.

[0627] Embodiment 138. The compound has the formula (VII-a):

[0628] [ka] 132. The compound of embodiment 131, wherein

[0629] Embodiment 139. The compound of any one of embodiments 131 to 137, wherein the compound is a solvate.

[0630] Embodiment 140. The compound of embodiment 139, wherein the compound is a solvate formed from any one of a mixture of acetonitrile and methyl tert-butyl ether, a mixture of nitromethane and methyl tert-butyl ether, a mixture of tetrahydrofuran and methyl tert-butyl ether, methyl acetate, and ethyl acetate.

[0631] Embodiment 141. The compound of any one of embodiments 131 to 137, wherein the compound is crystalline.

[0632] Embodiment 142. The compound of embodiment 141, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 4.8° ±0.2 degrees in combination with one or more peaks selected from 5.6°, 6.2°, 14.8°, and 15.6°.

[0633] Embodiment 143. The compound of embodiment 141, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.3° ±0.2 degrees in combination with one or more peaks selected from 7.7°, 10.5°, 11.3°, 11.6°, and 14.4°.

[0634] Embodiment 144. The compound of embodiment 141, wherein the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 6.2° and 6.9° ±0.2 degrees.

[0635] Embodiment 145. A method of synthesizing a compound according to embodiment 131, comprising: -Formula(H(dnp) 保護 ) and compounds of formula (Aib 保護 ) to form a compound of formula (H(dnp) 保護 -Aib 保護 forming a compound of

[0636] [ka] (In the formula, R 28 and R 31 is a protecting group) -Protecting group R 31 By removing (H(dnp) 保護 -Aib),

[0637] [ka] -Formula(H(dnp) 保護 -Aib) and the compound of formula (Q 保護2 ) to form a compound of formula (H(dnp) 保護 -Aib-Q 保護2 forming a compound of

[0638] [ka] (In the formula, R 29 and R32 is a protecting group) -Protecting group R 32 By removing (H(dnp) 保護 -Aib-Q 保護1 forming a compound of

[0639] [ka] -Formula(H(dnp) 保護 -Aib-Q 保護1 ) and compounds of formula (G 保護 with a compound of formula (VII) to form a compound of formula (VII),

[0640] [ka] (In the formula, R 30 is a protecting group), and optionally one or more protecting groups R 28 , R 29 , and R 30 The method of claim 1, further comprising:

[0641] Embodiment 146. The method of embodiment 145, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0642] Embodiment 147. A method for synthesizing a polypeptide of SEQ ID NO: 16, comprising conjugating a compound according to any one of embodiments 131 to 137 to the N-terminus of a polypeptide of SEQ ID NO: 17 via the C-terminus of the compound.

[0643] Embodiment 148. A compound of formula (VIII), or a salt, solvate, or hydrate thereof, wherein

[0644] [ka] In the formula, R 33 is H or a protecting group, and R 34 is H or a protecting group, and R35 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0645] Embodiment 149. The compound of embodiment 148, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0646] Embodiment 150.R 33 The compound according to any one of embodiments 148-149, wherein is H or Boc.

[0647] Embodiment 151.R 34 The compound according to any one of embodiments 148-150, wherein is H or trityl.

[0648] Embodiment 152.R 35 The compound according to any one of embodiments 148-151, wherein is H or tert-butyl.

[0649] Embodiment 153.R 33 , R 34 , and R 35 The compound of any one of embodiments 148-152, wherein at least one of: is H.

[0650] Embodiment 154.R 33 , R 34 , and R 35 The compound of any one of embodiments 148-153, wherein at least one of is a protecting group.

[0651] Embodiment 155. The compound has the formula (VIII-a):

[0652] [ka] The compound of embodiment 148, wherein

[0653] Embodiment 156. The compound of any one of embodiments 148-155, wherein the compound is a solvate.

[0654] Embodiment 157. The compound of embodiment 156, wherein the compound is a solvate formed from any one of a mixture of tetrahydrofuran and methyl tert-butyl ether, a mixture of tetrahydrofuran and heptane, a mixture of 1,4-dioxane and water, a mixture of ethyl acetate and methyl tert-butyl ether, and a mixture of acetonitrile and methyl tert-butyl ether.

[0655] Embodiment 158. The compound of any one of embodiments 148 to 155, wherein the compound is crystalline.

[0656] Embodiment 159. The compound of embodiment 158, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 4.7° ±0.2 degrees in combination with one or more peaks selected from 5.5°, 8.2°, 10.1°, 11.8°, 13.3°, 13.6°, and 18.9°; or

[0657] Embodiment 160. The compound of embodiment 158, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.8° ±0.2 degrees in combination with one or more peaks selected from 5.3°, 8.9°, 9.2°, 15.2°, 18.6°, and 19.5°.

[0658] Embodiment 161. A method of synthesizing a compound according to embodiment 148, comprising: -Formula(H(trt) 保護 ) and compounds of formula (Aib 保護 ) to form a compound of formula (H(trt) 保護 -Aib 保護 forming a compound of

[0659] [ka] (In the formula, R 33 and R 36is a protecting group) -Protecting group R 36 By removing (H(trt) 保護 -Aib),

[0660] [ka] -Formula(H(trt) 保護 -Aib) and the compound of formula (Q 保護2 ) to form a compound of formula (H(trt) 保護 -Aib-Q 保護2 forming a compound of

[0661] [ka] (In the formula, R 33 and R 37 is a protecting group) -Protecting group R 37 By removing (H(trt) 保護 -Aib-Q 保護1 forming a compound of

[0662] [ka] -Formula(H(trt) 保護 -Aib-Q 保護1 ) and compounds of formula (G 保護 to form a compound of formula (VIII),

[0663] [ka] (In the formula, R 35 is a protecting group), and optionally one or more protecting groups R 33 , R 34 , and R 35 The method of claim 1, further comprising:

[0664] Embodiment 162. The method of embodiment 161, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0665] Embodiment 163. A method for synthesizing a polypeptide of SEQ ID NO: 16, comprising conjugating a compound according to any one of embodiments 148 to 155 to the N-terminus of a polypeptide of SEQ ID NO: 17 via the C-terminus of the compound.

[0666] Embodiment 164. A compound of formula (IX) or a salt, solvate, or hydrate thereof,

[0667] [ka] In the formula, R 38 is H or a protecting group, and R 39 is H or a protecting group, and R 40 is H or a protecting group, and R 41 is H or a protecting group, and R 42 is H or a protecting group, and R 43 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0668] Embodiment 165. The compound of embodiment 164, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0669] Embodiment 166.R 38 The compound according to any one of embodiments 164-165, wherein is H or Fmoc.

[0670] Embodiment 167.R 39 The compound according to any one of embodiments 164-166, wherein is H or tert-butyl.

[0671] Embodiment 168.R 40 The compound according to any one of embodiments 164-167, wherein is H or tert-butyl.

[0672] Embodiment 169.R 41 The compound according to any one of embodiments 164-168, wherein is H or tert-butyl.

[0673] Embodiment 170.R 42 The compound according to any one of embodiments 164-169, wherein is H or Boc.

[0674] Embodiment 171.R 43 The compound according to any one of embodiments 164-170, wherein is H or tert-butyl.

[0675] Embodiment 172.R 38 , R 39 , R 40 , R 41 , R 42 , and R 43 The compound of any one of embodiments 164-171, wherein at least one of: is H.

[0676] Embodiment 173.R 38 , R 39 , R 40 , R 41 , R 42 , and R 43 The compound of any one of embodiments 164-172, wherein at least one of is a protecting group.

[0677] Embodiment 174. The compound has the formula (IX-a):

[0678] [ka] The compound of embodiment 164, wherein

[0679] Embodiment 175. The compound of any one of embodiments 164-174, wherein the compound is a solvate.

[0680] Embodiment 176. The compound of embodiment 175, wherein the compound is a solvate formed from any one of a mixture of methyl acetate and dibutyl ether, a mixture of acetone and dibutyl ether, a mixture of acetonitrile and dibutyl ether, a mixture of ethyl acetate and dibutyl ether, a mixture of methyl acetate and heptane, and a mixture of methyl ethyl ketone and dibutyl ether.

[0681] Embodiment 177. The compound of any one of embodiments 164-174, wherein the compound is a desolvate or anhydrous.

[0682] Embodiment 178. The compound of any one of embodiments 164-174, wherein the compound is crystalline.

[0683] Embodiment 179. The compound of embodiment 178, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.3° ±0.2 degrees in combination with one or more peaks selected from 6.0°, 6.9°, 7.2°, 8.0°, 12.2°, and 15.6°.

[0684] Embodiment 180. The compound of embodiment 178, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.8° ±0.2 degrees in combination with one or more peaks selected from 4.4°, 6.6°, 10.1°, 11.4°, 13.4°, and 15.5°.

[0685] Embodiment 181. The compound of embodiment 178, wherein the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 4.5° and 5.5° ±0.2 degrees, in combination with one or more peaks selected from 6.0° and 7.3°.

[0686] Embodiment 182. A method of synthesizing a compound according to embodiment 164, comprising: -Formula(D 保護 ) and a compound of formula (Y保護2 ) to react with a compound of formula (D 保護 -Y 保護2 forming a compound of

[0687] [ka] (In the formula, R 38 , R 39 , R 40 , and R 44 is a protecting group) -Protecting group R 44 Remove (D 保護 -Y 保護1 forming a compound of

[0688] [ka] -Formula(D 保護 -Y 保護1 ) and compounds of formula (S 保護2 ) to react with a compound of formula (D 保護 -Y 保護1 -S 保護2 forming a compound of

[0689] [ka] (In the formula, R 41 and R 45 is a protecting group) -Protecting group R 45 Remove (D 保護 -Y 保護1 -S 保護1 forming a compound of

[0690] [ka] -Formula(D 保護 -Y 保護1 -S 保護1 ) and compounds of formula (K 保護2 ) to form a compound of formula (IX),

[0691] [ka] (In the formula, R 42 and R 43 is a protecting group), and optionally one or more protecting groups R 38 , R 39 , R 40 , R 41 , R 42 , and R 43 The method of claim 1, further comprising:

[0692] Embodiment 183. The method of embodiment 182, wherein the protecting group is selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0693] Embodiment 184. A method for synthesizing a polypeptide of SEQ ID NO: 16, comprising: (i) conjugating a compound described in any one of embodiments 164 to 174 via the C-terminus of the compound to the N-terminus of a polypeptide of SEQ ID NO: 18; and (ii) conjugating the compound via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO: 19.

[0694] Embodiment 185. A compound of Formula (X) or a salt, solvate, or hydrate thereof,

[0695] [ka] In the formula, R 46 is H or a protecting group, and R 47 is H or a protecting group, and R 48 is H or a protecting group, and R 49 is H or a protecting group, and R 50 is H or a protecting group, and R 51 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0696] Embodiment 186. The compound of embodiment 185, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0697] Embodiment 187.R 46 The compound according to any one of embodiments 185-186, wherein is H or tert-butyl.

[0698] Embodiment 188.R 47 The compound according to any one of embodiments 185-187, wherein is H or tert-butyl.

[0699] Embodiment 189.R 48 The compound according to any one of embodiments 185-188, wherein is H or Boc.

[0700] Embodiment 190.R 49 The compound according to any one of embodiments 185-189, wherein is H or Fmoc.

[0701] Embodiment 191.R 50 The compound according to any one of embodiments 185-190, wherein is H or tert-butyl.

[0702] Embodiment 192.R 51 The compound according to any one of embodiments 185-191, wherein is H or tert-butyl.

[0703] Embodiment 193.R 46 , R 47 , R 48 , R 49 , R 50 , and R 51 The compound of any one of embodiments 185-192, wherein at least one of: is H.

[0704] Embodiment 194.R 46 , R 47 , R 48 , R 49 , R 50 , and R 51 The compound of any one of embodiments 185-193, wherein at least one of is a protecting group.

[0705] Embodiment 195. The compound has the formula (Xa):

[0706] [ka] The compound of embodiment 185, wherein

[0707] Embodiment 196. The compound of any one of embodiments 185-195, wherein the compound is a solvate.

[0708] Embodiment 197. The compound of embodiment 196, wherein the compound is a solvate formed from ethanol or isopropyl alcohol.

[0709] Embodiment 198. The compound of any one of embodiments 185 to 195, wherein the compound is a desolvate.

[0710] Embodiment 199. The compound of any one of embodiments 185-195, wherein the compound is crystalline.

[0711] Embodiment 200. The compound of embodiment 199, wherein the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 18.1° and 18.7° ±0.2 degrees, in combination with one or more peaks selected from 5.7°, 8.7°, 13.7°, 14.3°, 15.9°, and 16.2°.

[0712] Embodiment 201. The compound of embodiment 199, wherein the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 5.9° and 10.5° ±0.2 degrees, in combination with one or more peaks selected from 7.1°, 8.9°, 14.6°, and 16.6°.

[0713] Embodiment 202. The compound of embodiment 199, wherein the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 7.8° and 20.3° ±0.2 degrees, in combination with one or more peaks selected from 5.8°, 15.5°, and 19.5°.

[0714] Embodiment 203. The compound of embodiment 199, wherein the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 5.9° and 7.4° ±0.2 degrees, in combination with one or more peaks selected from 6.5°, 6.9°, and 14.8°.

[0715] Embodiment 204. A method of synthesizing a compound according to embodiment 185, comprising: -Formula (Y 保護 ) and compounds of formula (S 保護2 ) to react with a compound of formula (Y 保護 -S 保護2 forming a compound of

[0716] [ka] (In the formula, R 46 , R 47 , R 48 , and R 52 is a protecting group) -Protecting group R 52 By removing (Y 保護 -S 保護1 forming a compound of

[0717] [ka] -Formula (Y 保護 -S 保護1 ) and compounds of formula (K 保護2 ) to react with a compound of formula (Y 保護 -S 保護1 -K 保護2 forming a compound of

[0718] [ka] (In the formula, R 49 and R 53 is a protecting group) -Protecting group R 53 By removing (Y 保護 -S 保護1 -K 保護1 forming a compound of

[0719] [ka] -Formula (Y 保護 -S 保護1 -K 保護1 ) and a compound of formula (Y 保護2 ) to form a compound of formula (X),

[0720] [ka] (In the formula, R 50 and R 51 is a protecting group), and optionally one or more protecting groups R 46 , R 47 , R 48 , R 49 , R 50 , and R 51 The method of claim 1, further comprising:

[0721] Embodiment 205. The method of embodiment 204, wherein the protecting group is selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0722] Embodiment 206. A method for synthesizing a polypeptide of SEQ ID NO: 16, comprising: (i) conjugating a compound described in any one of embodiments 185 to 195 via the C-terminus of the compound to the N-terminus of a polypeptide of SEQ ID NO: 20; and (ii) conjugating the resulting compound via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO: 21.

[0723] Embodiment 207. A compound of formula (XI) or a salt, solvate, or hydrate thereof,

[0724] [ka] In the formula, R 64 is H or a protecting group, and R 65 is H or a protecting group, and R 66 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0725] Embodiment 208. The compound of embodiment 207, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0726] Embodiment 209.R 64 , R 65 , and R 66 The compound of any one of embodiments 207-208, wherein at least one of: is H.

[0727] Embodiment 210.R 64 , R 65 , and R 66 The compound of any one of embodiments 207-209, wherein at least one of is a protecting group.

[0728] Embodiment 211. The compound has the formula:

[0729] [ka] 208. The compound of embodiment 207, wherein

[0730] Embodiment 212. The compound of any one of embodiments 207-211, wherein the compound is a solvate.

[0731] Embodiment 213. The compound of any one of embodiments 207-211, wherein the compound is crystalline.

[0732] Embodiment 214. The compound of embodiment 213, wherein the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 6.1° and 8.5° ±0.2 degrees, in combination with one or more peaks selected from 5.8°, 16.9°, 18.5°, 18.8°, 19.3°, and 20.9°.

[0733] Embodiment 215. A compound of formula (XII) or a salt, solvate, or hydrate thereof,

[0734] [ka] In the formula, R 61 is H or a protecting group, and R 62 is H or a protecting group, and R 63 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0735] Embodiment 216. The compound of embodiment 215, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0736] Embodiment 217.R 61 The compound according to any one of embodiments 215-216, wherein is H or tert-butyl.

[0737] Embodiment 218.R 62 The compound according to any one of embodiments 215-217, wherein is H or tert-butyl.

[0738] Embodiment 219.R 63 The compound according to any one of embodiments 215-218, wherein is H or tert-butyl.

[0739] Embodiment 220.R 61 , R 62 , and R 63 The compound of any one of embodiments 215-219, wherein at least one of is H.

[0740] Embodiment 221.R 61 , R 62 , and R 63 The compound of any one of embodiments 215-220, wherein at least one of is a protecting group.

[0741] Embodiment 222. The compound has the formula (XII-a):

[0742] [ka] 216. The compound of embodiment 215, wherein

[0743] Embodiment 223. The compound of any one of embodiments 215 to 222, wherein the compound is a solvate.

[0744] Embodiment 224. The compound of any one of embodiments 215 to 222, wherein the compound is a desolvate or anhydrous.

[0745] Embodiment 225. The compound of any one of embodiments 215 to 222, wherein the compound is crystalline.

[0746] Embodiment 226. The compound of embodiment 225, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 11.4° ±0.2 degrees in combination with one or more peaks selected from 6.0°, 8.9°, 12.7°, 13.6°, 14.6°, 17.0°, and 18.8°.

[0747] Embodiment 227. The compound of embodiment 225, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 10.6° ±0.2 degrees in combination with one or more peaks selected from 7.1°, 12.1°, 13.6°, 14.2°, 15.2°, 16.0°, and 16.8°.

[0748] Embodiment 228. The compound of embodiment 225, wherein the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta ±0.2 degrees of 10.1° and 15.5° in combination with one or more peaks selected from 6.1°, 8.7°, 11.4°, 16.6°, and 19.2°.

[0749] Embodiment 229. A compound of formula (XIII) or a salt, solvate, or hydrate thereof,

[0750] [ka] In the formula, R 67 is H or a protecting group, and R 68 is H or a protecting group, and R 69 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0751] Embodiment 230. The compound of embodiment 229, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0752] Embodiment 231.R 67 The compound according to any one of embodiments 229-230, wherein is H or tert-butyl.

[0753] Embodiment 232.R 68 The compound according to any one of embodiments 229-231, wherein is H or tert-butyl.

[0754] Embodiment 233.R 69 The compound according to any one of embodiments 229-232, wherein is H or tert-butyl.

[0755] Embodiment 234.R 67 , R 68 , and R 69 The compound of any one of embodiments 229-233, wherein at least one of: is H.

[0756] Embodiment 235.R 67 , R 68 , and R 69 The compound of any one of embodiments 229-234, wherein at least one of is a protecting group.

[0757] Embodiment 236. The compound has the formula:

[0758] [ka] 230. The compound of embodiment 229, wherein

[0759] Embodiment 237. The compound of any one of embodiments 229-236, wherein the compound is a solvate.

[0760] Embodiment 238. The compound of any one of embodiments 229 to 236, wherein the compound is a desolvate or anhydrous.

[0761] Embodiment 239. The compound of any one of embodiments 229-236, wherein the compound is crystalline.

[0762] Embodiment 240. The compound of embodiment 239, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.0° ±0.2 degrees in combination with one or more peaks selected from 8.3, 9.7, and 11.2°.

[0763] Embodiment 241. The compound of embodiment 239, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.2° ±0.2 degrees in combination with one or more peaks selected from 5.3, 8.1, 14.4, and 16.2°.

[0764] Embodiment 242. A compound of formula (XIV) or a salt, solvate, or hydrate thereof,

[0765] [ka] In the formula, R 70 is H or a protecting group, and R 71 is H or a protecting group, and R 72 is H or a protecting group, and R 73 is H or a protecting group, and R 74 is H or a protecting group, or a salt, solvate, or hydrate thereof.

[0766] Embodiment 243. The compound of embodiment 242, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

[0767] Embodiment 244.R 70 The compound according to any one of embodiments 242-243, wherein is H, Fmoc, or tert-butyl.

[0768] Embodiment 245.R 71 The compound according to any one of embodiments 242-244, wherein is H or tert-butyl.

[0769] Embodiment 246.R 72 The compound according to any one of embodiments 242 to 245, wherein is H or tert-butyl.

[0770] Embodiment 247.R 73 The compound according to any one of embodiments 242 to 246, wherein is H or tert-butyl.

[0771] Embodiment 248.R 74 The compound according to any one of embodiments 242 to 247, wherein is H, Fmoc, or tert-butyl.

[0772] Embodiment 249.R 70 , R 71 , R 72 , R 73 , and R 74The compound of any one of embodiments 242-248, wherein at least one of is H.

[0773] Embodiment 250.R 70 , R 71 , R 72 , R 73 , and R 74 The compound of any one of embodiments 242-249, wherein at least one of is a protecting group.

[0774] Embodiment 251. The compound has the formula:

[0775] [ka] 243. The compound of embodiment 242, wherein

[0776] Embodiment 252. The compound of any one of embodiments 242 to 251, wherein the compound is a solvate.

[0777] Embodiment 253. The compound of any one of embodiments 242 to 251, wherein the compound is a desolvate or anhydrous.

[0778] Embodiment 254. The compound of any one of embodiments 242 to 251, wherein the compound is crystalline.

[0779] Embodiment 255. The compound of embodiment 254, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.5° ±0.2 degrees in combination with one or more peaks selected from 6.1, 8.7, 10.6, 15.0, 16.1, and 18.6°. [Example]

[0780] Example 1: Boc-l-Tyr(tBu)-Aib-Glu(OtBu)-Gly-OH

[0781] [ka]

[0782] Compound 1 ((S)-3-(4-(tert-butoxy)phenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid) was added to 10 volumes (vol) of EtOAc and the temperature was adjusted to approximately -10 to 0 °C. BOP (1.3 equivalents) was added portionwise at -10 to 0 °C. Compound 2 (benzyl 2-amino-2-methylpropanoate), 1.3 equivalents, was added at -10 to 0 °C. DIPEA was added dropwise at -10 to 0 °C, and the mixture was stirred at -10 to 0 °C for 16 to 20 h. The mixture was warmed to 0 to 20 °C and washed five times with 10 volumes at 0 to 20 °C. The aqueous layers were combined. The organic layer was washed twice with 0.5 mol / L Na2CO3 (5 volumes) at 0 to 20 °C. The organic layer was washed with water (10 volumes) at 0 to 20 °C, and the aqueous layers were combined. The organic layer was concentrated to dryness below 40°C. 2 volumes of EtOAc were added at 15-25°C, and the mixture was stirred for 1 hour, then filtered to obtain a wet cake. The filtrate was concentrated to dryness below 40°C. 5 volumes of EtOH were added to form a clear solution, and the temperature was adjusted to 5-15°C. 1-2 volumes of water were added dropwise to the mixture, seed crystals (0.5% w / w) were added, and the mixture was stirred for 1-2 hours. 3-4 volumes of water were added dropwise to the mixture, and the mixture was stirred for 16-24 hours, then filtered. The wet cake was washed with EtOH / water (1 / 1 v / v). The wet cake was dried in vacuo at 35-45°C for 20-24 hours to obtain Preparation 1.

[0783] One equivalent of Preparation 1 was mixed with Pd / C (0.1x). MeOH (10.5x, 13.3 vol) was added to the mixture, and the mixture was exchanged for argon three times, followed by hydrogen three times. The pressure was adjusted to 45 psi with hydrogen, and the mixture was heated to 40°C and stirred for 18-24 hours. The mixture was cooled to 20-30°C, the argon was exchanged three times, and the mixture was filtered. The wet cake was washed with MeOH (1 vol), and the filtrate was transferred to a container and concentrated to approximately 2 volumes below 45°C. EA (300 ml, 3 vol) was added, and the mixture was concentrated to approximately 2 volumes below 45°C. EA (300 ml, 3 vol) was added, and the mixture was concentrated to approximately 2 volumes below 45°C. Heptane (8 vol) was added dropwise, and the mixture was stirred at 20-35°C for 20-24 hours, after which the mixture was filtered. The wet cake was washed with heptane (1 volume) and then dried in vacuo at 40-50°C for 20-24 hours to give Preparation 2.

[0784] THF was added to Vessel 1 and cooled to 0°C. IBCF (1.3 equiv., 0.42x) was mixed with Preparation 2 (1.0x). 3 volumes of THF were added to a separate vessel 2, and NMM (1.35 equiv., 0.325x) was added. This mixture was then added to the mixture containing Preparation 2 and stirred for 4-6 hours. 3 volumes of THF were added to Vessel 2, followed by 3 volumes of water, and the mixture was stirred at 15-20°C for 30 minutes. The contents of Vessel 2 were added to Vessel 1 at 0-5°C. Vessel 1 was heated to 20°C over 3-4 hours and stirred for 16-20 hours. 15 volumes of EA were added to Vessel 1, and the organic layer was separated. 5% aqueous KHSO4 (16 volumes) was added to the organic layer at 15-25°C, and the mixture was stirred for 1-2 hours. The organic layer was separated and washed twice with 5% NaCl (10 volumes). The mixture in vessel 1 was concentrated to approximately 1 volume below 45°C, THF (5 volumes) was added, the mixture was concentrated to approximately 1 volume, IPA / HO 1:2 (10.5 volumes) was added, and the mixture was stirred for 48 hours at 15-25°C. The mixture was then filtered, the wet cake was washed twice with IPA / HO 1:2 (1 volume), and the wet cake was dried in vacuo at 40-45°C for 20-24 hours to produce Preparation 3.

[0785] Preparation 3 (1.0x) was added to benzyl glycinate (1.3 equiv, 0.44x), followed by 10 volumes of ACN. 2,6-Lutidine (3.0 equiv, 0.53x) was added, and the temperature was adjusted to -10°C. COMU (1.3 equiv, 0.86x) was added at -10°C, and the mixture was stirred for 2-4 hours. The mixture was concentrated to approximately 1 volume below 45°C, EA (10 volumes) was added, and the mixture was stirred at 15-20°C for 0.5-1 hour. The mixture was then filtered, and the wet cake was washed with EA (1 volume). The filtrate was added and concentrated to approximately 1 volume below 45°C. MTBE (10 volumes) was added, and the mixture was stirred at 15-20°C for 0.5-1 hour. The mixture was then filtered, and the wet cake was washed with MTBE (1 volume). The filtrate was washed twice with 0.5 ml / L aqueous NaCO (10 volumes) at 15-25°C, twice with 5% aqueous KHSO (10 volumes) at 15-25°C, and with water (10 volumes) at 15-25°C. The organic layer was separated and concentrated to 1-2 volumes below 45°C. MTBE (10 volumes) was added, and the mixture was heated to 40°C and cooled to 10°C over 2 hours. The mixture was filtered, and the cake was dried at 45°C for 16-24 hours to produce Preparation 4.

[0786] Preparation 4 (1×, 1 equiv.) was added to IPA (7.92×, 10 vol.), charged with Pd / C (0.1×), and the mixture was flushed with argon three times, then with hydrogen three times. The pressure was adjusted to 45 psi with hydrogen, and the mixture was heated to 40°C and stirred for 18–24 h. The mixture was cooled to 20–30°C, the argon was flushed three times, the mixture was filtered, the wet cake was washed with IPA (1 vol.), and the filtrate was concentrated to approximately 1 vol. at 45°C. MTBE (3 vol.) was added, and the mixture was concentrated to approximately 1 vol. below 45°C. MTBE (10 vol.) and IPA (0.5 vol.) were added to the mixture, and the mixture was heated to 40°C, cooled to 20°C over 2 h, and stirred at 20°C for 2–6 h. The solid was filtered, the wet cake was washed with 1 volume of MTBE, and the wet cake was dried under vacuum at 40-50°C for 16-24 hours to produce Y-Aib-EG.

[0787] Preparation of Y-Aib-EG Forms A, B, and C Three solid forms of the Y-Aib-EG tetramer were prepared: Form A, Form B, and Form C. Y-Aib-EG Form A is a solvated form produced from MTBE or mixtures containing MTBE (e.g., MTBE / IPA, MTBE / heptane). Y-Aib-EG Form B is a desolvated form from Form A by drying at high temperature / vacuum, heating to 110°C, or exposure to 75% RH. Y-Aib-EG Form C is a hydrate containing various amounts of water in the unit cell, including zero water content.

[0788] Crystallization of Y-Aib-EG helps eliminate impurities from the crude material. Compared to amorphous crude Y-Aib-EG, crystalline Y-Aib-EG shows improved purity based on UPLC-MS analysis. Crystallization of crude Y-Aib-EG spiked with 1% dimer (impurity) yielded crystalline Y-Aib-EG with almost complete elimination of the dimer. Crystallization also improves physical properties, such as hygroscopicity. Dynamic vapor sorption (DVS) analysis showed that amorphous Y-Aib-EG showed a 4.1 wt% increase in sorption from 0% to 95% RH, whereas Y-Aib-EG Form B only increased by 1.4 wt% under the same conditions.

[0789] XRPD patterns of crystalline TZP tetramer were obtained on a Bruker D8 Endeavor X-ray powder diffractometer equipped with a CuKα (1.5418 Å) source and a Linxeye detector, operating at 40 kV and 40 mA. Samples were scanned from 4 to 42° 2θ using a 0.009° 2θ step size and a 0.5 s / step scan rate, using a 0.3° primary slit aperture and a 3.9° particle size distribution (PSD) aperture. In some cases, samples were scanned from 4 to 30° 2θ at a 0.25 s / step scan rate. The powder was packed into a quartz sample holder, and a smooth surface was obtained using a glass slide. Crystalline form diffraction patterns were collected at ambient temperature and relative humidity. Crystalline peak positions were determined using MDI-Jade v7.9.9. It is well known in the field of crystallography that for any crystalline form, the relative intensities of diffraction peaks may vary due to preferred orientation resulting from factors such as crystalline form and crystal habit. When the effect of preferred orientation is present, peak intensities change, but the characteristic peak positions of a polymorph remain unchanged. See, for example, The United States Pharmacopeia #23, National Formulary #18, pages 1843-1844, 1995. It is also well known in the field of crystallography that for any crystalline form, angular peak positions may vary slightly. For example, peak positions may shift due to variations in the temperature at which the sample is analyzed, sample displacement, or the presence or absence of an internal standard. In this case, a peak position variation of ±0.2 2θ° is estimated to account for these potential variations without precluding unambiguous identification of the indicated crystalline form. Confirmation of a crystalline form can be based on any unique combination of characteristic peaks.

[0790] Y-Aib-EG Form A 0.2 mL MTBE was added to 110.9 mg of the amorphous solid Y-Aib-EG, and the sample was stirred at ambient conditions. After 4 days, an additional 0.4 mL of MTBE was added to the mixture, and the sample was stirred for another day. A white slurry was obtained, which was transferred to a 0.45 μm nylon centrifuge filter and centrifuged at ambient temperature for 5 minutes. The resulting white solid was consistent with Y-Aib-EG Form A.

[0791] The prepared sample of Y-Aib-EG Form A is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as shown in Table 1 below, specifically, a peak at 5.2° 2-theta in combination with one or more peaks selected from 9.9, 10.4, 15.5, and 17.1° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of Y-Aib-EG Form A is shown in Figure 1A.

[0792] [Table 1]

[0793] Y-Aib-EG form B 6 mL MTBE was added to 1.01 grams of the amorphous solid Y-Aib-EG, and the sample was stirred at ambient conditions for 5 days to obtain a white slurry. The slurry was transferred to a 0.45 μm nylon centrifuge tube filter and centrifuged at ambient temperature for 5 minutes to obtain a white solid (0.78 g), which was primarily consistent with Y-Aib-EG Form A. 81.6 mg of such a white solid was dried under vacuum at approximately 50°C for 1 day. The resulting white solid was consistent with Y-Aib-EG Form B.

[0794] The prepared sample of Y-Aib-EG Form B is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 2 below, specifically, a peak at 6.1° 2-theta in combination with one or more peaks selected from 10.3, 16.8, and 18.1° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of Y-Aib-EG Form B is shown in Figure 1B.

[0795] [Table 2]

[0796] Y-Aib-EG form C 285.8 mg of amorphous Y-Aib-EG was stirred overnight at ambient conditions in 1 mL of water to obtain a thick slurry. Additional amounts of water (2 x 0.5 mL) and IPA (2 x 0.1 mL) were added to the slurry, and the sample was stirred at ambient conditions for an additional 6 days. The resulting off-white slurry was transferred to a 0.45 μm nylon centrifuge tube filter and centrifuged at ambient temperature for 5 minutes. The isolated solid was white and moist and was dried under vacuum at 70-78 °C for 2 hours. The resulting white solid was consistent with Y-Aib-EG Form C.

[0797] The prepared sample of Y-Aib-EG Form C is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 3 below, specifically, a peak at 6.2° 2-theta in combination with one or more peaks selected from 9.5, 13.6, and 21.8° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of Y-Aib-EG Form C is shown in Figure 1C.

[0798] [Table 3]

[0799] Coupling of Y-Aib-EG tetramer to a decamer for TZP synthesis

[0800] [ka]

[0801] Crystalline Y-Aib-EG can be used as a starting material to generate the TZP fragment (AA1-14) by coupling with the fragment decamer (AA5-14). The TZP fragment (AA1-14) can then be coupled with other fragments to generate the protected TZP as the active pharmaceutical ingredient (API).

[0802] The TZP fragment 10-mer (AA5-14) protected on resin (1.2518 g, 0.500 mmol) in a 45 ml automated synthesis reactor was swollen with 3 × 15 ml DMF for 15 min each, deprotected with 3 × 15 ml 20% piperidine / DMF for 30 min each, and washed with 5 × 15 ml DMF for 1 min each. A solution of TZP tetramer (1.055 g, 1.500 mmol, 94.53 wt%) and ethyl cyanoglyoxylate-2-oxime (214.3 mg, 1.500 mmol, 99 wt%) in 9 ml DMF was prepared. N,N'-Diisopropylcarbodiimide (258 μL, 1.650 mmol, 100 wt%) was added, and the resulting yellow solution was allowed to stand for 30 min with occasional shaking. The solution was then added to the reactor containing the resin. The reaction was mixed at ambient temperature for 18 hours and then drained. The resin was washed with 5 x 15 ml DMF for 1 minute each and 5 x 15 ml DCM for 1 minute each, then drained and dried for 4 hours to a constant weight of 1.4077 g.

[0803] For analysis, approximately 300 mg of resin was treated with 5 mL of a solution consisting of trifluoroacetic acid (4.64 mL, 61.4 mmol, 100% by weight), triisopropylsilane (125 μL, 0.609 mmol, 100% by weight), water (125 μL, 6.93874 mmol, 100% by weight), and DTT (125 mg, 0.810357 mmol, 100% by weight). The mixture was stirred on a rotary mixer for 2 hours, filtered, and washed with trifluoroacetic acid (2 mL, 26.45 mmol, 100% by weight). The combined filtrate and wash were added to 35 mL of cold MTBE in a centrifuge tube. After 30 minutes in the freezer, the mixture was centrifuged and the supernatant was decanted. The residual solid was washed with 2 × 30 mL of room-temperature MTBE in a centrifuge and dried overnight in a vacuum oven at 35 °C. The yield of solid was 154.7 mg.

[0804] Analysis of the isolated solid by LC / MS showed complete coupling of the 10-mer to the tetramer, producing the TZP fragment 14-mer (AA1-14).

[0805] Example 2: Fmoc-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-OH

[0806] [ka]

[0807] Resin swelling: The resin (0.500 mmol) was added to the reactor and allowed to swell with DMF (3 x 10 mL x 20 min).

[0808] Washing after Fmoc removal: After deprotection, the resin was washed with DMF (5 x 10 mL x 2 minutes).

[0809] Post-coupling wash: After coupling, the resin was washed with DMF (5 x 10 mL x 2 min).

[0810] Resin washing and drying: After the last coupling or deprotection, the resin was washed with DMF (5 x 10 mL x 2 min), followed by DCM (5 x 10 mL x 2 min) and evacuated to dryness under N2 atmosphere until constant weight.

[0811] [Table 4]

[0812] Four crystalline solid forms of the TFTS tetramer have been identified: Form A, Form B, Form C, and Form D. Form A is a crystalline material produced from organic solvent / heptane mixtures, such as IPA / heptane, EtOAc / heptane, MEK / heptane, iPrOAc / heptane, and THF / heptane. It represents a family of isostructural solvates. TFTS Form B is a semi-disordered crystalline material produced from MTBE. TFTS Form C is a semi-disordered crystalline material produced from ACN or 1:1 ACN / HO. TFTS Form D is a crystalline, desolvated or partially desolvated material from TFTS Form A with variable amounts of solvent in the unit cell.

[0813] The XRPD pattern of the crystalline TFTS tetramer was obtained on a Bruker D8 Endeavor X-ray powder diffractometer equipped with a CuKα (1.5418 Å) source and a Linxeye detector, operating at 40 kV and 40 mA. The sample was scanned from 4 to 30° 2θ with a step size of 0.009° 2θ and a scan rate of 0.25 s / step, using a 0.3° primary slit aperture and a 3.9° particle size distribution (PSD) aperture. The powder was packed into a quartz or silicon sample holder, and a smooth surface was obtained using a glass slide. The crystalline form diffraction pattern was collected at ambient temperature and relative humidity. Crystalline peak positions were determined using MDI-Jade v7.9.9. It is well known in the field of crystallography that for any crystalline form, the relative intensities of diffraction peaks may vary due to preferred orientation resulting from factors such as crystalline morphology and crystal habit. When preferred orientation effects exist, peak intensities change, but the characteristic peak positions of a polymorph remain unchanged. See, for example, The United States Pharmacopeia #23, National Formulary #18, pages 1843-1844, 1995. Furthermore, it is well known in the field of crystallography that angular peak positions may vary slightly for any crystalline form. For example, peak positions may shift due to variations in the temperature at which the sample is analyzed, sample displacement, or the presence or absence of an internal standard. In this case, a peak position variation of ±0.2 2θ° is estimated to account for these potential variations without precluding unambiguous identification of the indicated crystalline form. Confirmation of a crystalline form can be based on any unique combination of characteristic peaks.

[0814] TFTS ​​form A In one experiment, TFTS Form A was prepared by adding 0.5 mL of 1:1 vol / vol IPA / heptane to 104.8 mg of amorphous TFTS, followed by another 0.5 mL of heptane. The sample was stirred overnight at ambient conditions to obtain a thin slurry. The sample was uncapped and left at ambient conditions for 3 days to allow the solvent to evaporate. The resulting white solid was consistent with TFTS Form A.

[0815] In a separate experiment, TFTS Form A was prepared by stirring 62.1 mg of amorphous TFTS in 300 μL of 10:90 vol / vol MEK / heptane at 5 °C. A thick white slurry was obtained. After 5 days, the sample was centrifuged at ambient conditions, the liquid phase was decanted, and the resulting white solid was consistent with TFTS Form A.

[0816] A prepared sample of YFTS Form A is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 4 below, specifically, peaks at 4.3 and 12.8° 2-theta in combination with one or more peaks selected from 5.8, 7.7-7.9, 10.0, 10.8-10.9, 11.3-11.4, 12.0-12.1, and 21.8° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of YFTS Form A is shown in Figure 2A.

[0817] [Table 5]

[0818] TFTS ​​tetrameric form B 2×0.2 mL of MTBE was added to 100.2 mg of amorphous TFTS, and the sample was capped and stirred / vortexed at ambient conditions to give a thick white slurry. The wet solid from the slurry was consistent with TFTS Form B.

[0819] A prepared sample of TFTS form B is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 5 below, specifically peaks at 5.7 and 7.3° 2-theta in combination with one or more peaks selected from 5.1, 7.6, 9.5, and 12.4° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of TFTS tetramer form B is shown in Figure 2B.

[0820] [Table 6]

[0821] TFTS ​​form C In one experiment, 0.25 mL of 1:1 vol / vol ACN / HO was added to 89.6 mg of amorphous TFTS, the sample was capped, and stirred overnight at ambient conditions to give a thick white slurry. The slurry was centrifuged at ambient conditions for approximately 5 minutes, the liquid phase was decanted, and the resulting wet solid was consistent with TFTS Form C.

[0822] In a separate experiment, 3 × 0.1 mL of ACN was added to 56.6 mg of amorphous TFTS, the sample was capped, and stirred at ambient conditions for approximately 3 hours. The resulting wet solid from the thick white slurry was consistent with TFTS Form C.

[0823] A prepared sample of TFTS form C is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 6 below, specifically, a peak at 7.8° 2-theta in combination with one or more peaks selected from 8.5, 12.0, 15.5, 20.2, and 23.3° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of TFTS form C is shown in Figure 2C.

[0824] [Table 7]

[0825] TFTS ​​form D The TFTS Form A solid was air-dried at ambient conditions for at least 7 days to give TFTS Form D.

[0826] A prepared sample of TFTS form D is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 7 below, specifically peaks at 4.2 and 8.2-8.3 degrees 2-theta, in combination with one or more peaks selected from 5.9, 7.7, 9.2, 10.2, 11.3, 13.8-13.9, 15.5-15.7, 17.1, and 18.5 degrees 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of TFTS form D is shown in Figure 2D.

[0827] [Table 8]

[0828] Example 3: Pro-Ser(tBu)-Ser(tBu)-Gly-NH

[0829] [ka]

[0830] Example 3 was prepared substantially according to the procedure described in Example 4 below.

[0831] Resin swelling: The resin (0.500 mmol) was charged into a reactor and allowed to swell with DMF (3 x 10 mL x 20 min).

[0832] Washing after Fmoc removal: After deprotection, the resin was washed with DMF (5 x 10 mL x 2 minutes).

[0833] Post-coupling wash: After coupling, the resin was washed with DMF (5 x 10 mL x 2 min).

[0834] Resin washing and drying: After the last coupling or deprotection, the resin was washed with DMF (5 x 10 mL x 2 min), followed by DCM (5 x 10 mL x 2 min) and evacuated to dryness under N2 atmosphere until constant weight.

[0835] A total of seven crystalline solid forms of PSSG-NH2 have been identified, including PSSG-NH2 forms A through H. A polymorphic map illustrating their relationships is shown in the scheme in Figure 3.

[0836] Of these forms, PSSG-NH2 forms B and G are anhydrous / non-solvated, while C, D, E, F, and H are solvated. Forms A, E, and H are disordered crystalline materials.

[0837] XRPD patterns of crystalline PSSG-NH2 were obtained on a Bruker D8 Endeavor X-ray powder diffractometer equipped with a CuKα (1.5418 Å) source and a Linxeye detector, operating at 40 kV and 40 mA. Samples were scanned from 4 to 42 2θ° with a step size of 0.009 2θ° and a scan rate of 0.5 s / step, using a 0.3° primary slit aperture and a 3.9° particle size distribution (PSD) aperture. In some cases, samples were scanned from 4 to 30 2θ° at a scan rate of 0.25 s / step. Powders were packed into quartz or silicon sample holders, and smooth surfaces were obtained using glass slides. Crystalline form diffraction patterns were collected at ambient temperature and relative humidity. Crystalline peak positions were determined using MDI-Jade v7.9.9. It is well known in the field of crystallography that for any crystalline form, the relative intensities of diffraction peaks may vary due to preferred orientation resulting from factors such as crystalline form and crystal habit. When the effect of preferred orientation is present, peak intensities change, but the characteristic peak positions of a polymorph remain unchanged. See, for example, The United States Pharmacopeia #23, National Formulary #18, pages 1843-1844, 1995. It is also well known in the field of crystallography that for any crystalline form, angular peak positions may vary slightly. For example, peak positions may shift due to variations in the temperature at which the sample is analyzed, sample displacement, or the presence or absence of an internal standard. In this case, a peak position variation of ±0.2 2θ° is estimated to account for these potential variations without precluding unambiguous identification of the indicated crystalline form. Confirmation of a crystalline form can be based on any unique combination of characteristic peaks.

[0838] PSSG-NH2 Form A 12.7549 g of solid-phase peptide synthesis resin for PSSG-NH2 tetramer was loaded into a 150 mL fritted reactor equipped with an overhead stirrer. 128 mL of 5% TFA / DCM cleavage cocktail (6.4 mL TFA, 121.6 mL DCM) was charged to the reactor, and the resulting solution was stirred for 30 minutes. The reactor was drained and washed with DCM (2 × 120 mL). MTBE was added to the filtrate to precipitate the tetramer. The resulting precipitate was filtered through a Buchner funnel, air-dried under vacuum suction overnight, and then dried overnight in a vacuum oven at 35 °C to remove residual solvent. The resulting solid was consistent with PSSG-NH2 Form A.

[0839] A prepared sample of PSSG-NH2 Form A is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 8 below, specifically peaks at 7.0 and 8.0 degrees 2-theta in combination with one or more peaks selected from 10.3, 14.1, 16.7, and 19.0 degrees 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of PSSG-NH2 Form A is shown in Figure 4A.

[0840] [Table 9]

[0841] PSSG-NH2 Form B In one experiment, 80.9 mg of solid PSSG-NH2 Form A was stirred in 0.8 mL ACN at ambient conditions, resulting in a white suspension. After 8 days, the slurry was isolated by centrifuging the mixture at ambient conditions for 5 minutes using a centrifuge tube filter. The resulting white solid was consistent with PSSG-NH2 Form B.

[0842] In a separate experiment, 5.3 mg of solid PSSG-NH2 Form A was added to a clean TGA pan and heated to 150 °C on the TGA and held at that temperature for 60 minutes. The resulting off-white solid was consistent with PSSG-NH2 Form B.

[0843] In yet another experiment, 7.0 mg of solid PSSG-NH2 Form C was added to a clean TGA pan and heated to 165°C on the TGA and held at that temperature for 5 minutes. The resulting white solid was consistent with PSSG-NH2 Form B.

[0844] A prepared sample of PSSG-NH2 Form B is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 9 below, specifically, a peak at 9.0° 2-theta in combination with one or more peaks selected from 5.7, 9.9, 16.2, 18.1, and 18.4° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of PSSG-NH2 Form B is shown in Figure 4B.

[0845] [Table 10]

[0846] PSSG-NH2 Form C In one experiment, 83.6 mg of solid PSSG-NH2 Form A was stirred in 0.8 mL of wet EtOAc at ambient conditions, resulting in a white suspension. After 8 days, the slurry was isolated by centrifuging the mixture at ambient conditions for 5 minutes using a centrifuge tube filter. The resulting white solid was consistent with PSSG-NH2 Form C.

[0847] In another experiment, a mixture containing PSSG-NH2 Form C and Form D was air-dried overnight at ambient conditions to give a single phase of PSSG-NH2 Form C.

[0848] A prepared sample of PSSG-NH2 form C is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 10 below, specifically peaks at 5.6 and 10.5 degrees 2-theta in combination with one or more peaks selected from 11.9, 13.3, and 21.1 degrees 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of PSSG-NH2 form C is shown in Figure 4C.

[0849] [Table 11]

[0850] PSSG-NH2 Form D 76.0 mg of solid PSSG-NH2 Form A was stirred in 0.8 mL of EtOAc at ambient conditions to give a thick white slurry. The next day, an additional 0.5 mL of EtOAc was added, and the sample was stirred at ambient temperature for an additional 7 days. The mixture was then transferred to a centrifuge tube filter and centrifuged at ambient conditions for 5 minutes, and the solid was allowed to air-dry at ambient conditions for approximately 4 hours. A mixture of white solid and translucent mass was obtained. XRPD indicated that the sample consisted of Form C and Form D.

[0851] A prepared sample of PSSG-NH2 form D is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 11 below, specifically, a peak at 4.9° 2-theta in combination with one or more peaks selected from 10.7, 14.8, and 20.3° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of PSSG-NH2 form D is shown in Figure 4D.

[0852] [Table 12]

[0853] PSSG-NH2 Form E 66.9 mg of solid PSSG-NH2 Form A was stirred in 0.8 mL MeOAc at ambient conditions to give a white suspension. After 8 days, the slurry was isolated by centrifuging the mixture at ambient conditions for 5 minutes using a centrifuge tube filter. The solid was allowed to air dry at ambient temperature for approximately 4 hours to give a mixture of a white solid and translucent mass that exhibited an XRPD pattern consistent with PSSG-NH2 Form E.

[0854] A prepared sample of PSSG-NH2 form E is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 12 below, specifically peaks at 13.1 and 17.5 degrees 2-theta in combination with one or more peaks selected from 5.9, 10.5, 10.9, and 15.9 degrees 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of PSSG-NH2 form E is shown in Figure 4E.

[0855] [Table 13]

[0856] PSSG-NH2 form F 71.1 mg of solid PSSG-NH2 Form A was stirred in 0.8 mL THF at ambient conditions to give a white suspension. After 8 days, the slurry was isolated by centrifuging the mixture at ambient conditions for 5 minutes using a centrifuge tube filter, and the material was air-dried at ambient conditions for approximately 4 hours. The resulting white solid was consistent with PSSG-NH2 Form F.

[0857] A prepared sample of PSSG-NH2 form F is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 13 below, specifically, a peak at 7.8° 2-theta in combination with one or more peaks selected from 11.3, 11.5, 15.4, and 15.6° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of PSSG-NH2 form F is shown in Figure 4F.

[0858] [Table 14]

[0859] PSSG-NH2 form G 12.2 mg of solid PSSG-NH2 Form F was added to a clean TGA pan and heated to 125°C on the TGA and held at that temperature for 5 minutes. The resulting white solid was consistent with PSSG-NH2 Form G.

[0860] A prepared sample of PSSG-NH2 form G is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 14 below, specifically peaks at 10.0 and 12.5 degrees 2-theta in combination with one or more peaks selected from 8.1, 14.7, 18.8, and 22.4 degrees 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of PSSG-NH2 form G is shown in Figure 4G.

[0861] [Table 15]

[0862] PSSG-NH2 Form H 71.6 mg of solid PSSG-NH2 Form A was stirred in 0.8 mL IPA at ambient conditions to give a thick white slurry. The next day, an additional 0.5 mL IPA was added, and the sample was stirred at ambient temperature for an additional 7 days. The mixture was then transferred to a centrifuge tube filter and centrifuged at ambient conditions for 5 minutes, and the solid was allowed to air-dry at ambient conditions for approximately 4 hours. A mixture of white solid and translucent mass was obtained, which exhibited an XRPD pattern consistent with PSSG-NH2 Form H.

[0863] A prepared sample of PSSG-NH2 form H is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 15 below, specifically peaks at 10.8 and 21.1 degrees 2-theta in combination with one or more peaks selected from 5.6, 10.5, 11.9, and 15.4 degrees 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of PSSG-NH2 form H is shown in Figure 4H.

[0864] [Table 16]

[0865] Example 4: Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-NH

[0866] [ka]

[0867] Resin swelling: The resin (0.500 mmol) was charged into a reactor and allowed to swell with DMF (3 x 10 mL x 20 min).

[0868] Washing after Fmoc removal: After deprotection, the resin was washed with DMF (5 x 10 mL x 2 minutes).

[0869] Post-coupling wash: After coupling, the resin was washed with DMF (5 x 10 mL x 2 min).

[0870] Resin washing and drying: After the last coupling or deprotection, the resin was washed with DMF (5 x 10 mL x 2 min), followed by DCM (5 x 10 mL x 2 min) and evacuated to dryness under N2 atmosphere until constant weight.

[0871] [Table 17]

[0872] Cutting from resin: 5% TFA / DCM (10 volumes) was added to the resin and the reactor was agitated for 30 minutes. The reactor was drained and the resin was washed with DCM (2 x 5 volumes). The filtrate was added to pre-chilled MTBE:heptane (1:1, 10 volumes relative to the cleavage solution) and then centrifuged (3000 rpm x 10 minutes). The supernatant was discarded, fresh cold MTBE:heptane (5 volumes) was added, and the mixture was centrifuged (3000 rpm x 5 minutes). The supernatant was discarded, and the process was repeated once more with fresh MTBE:heptane. The supernatant was discarded, and the resulting material was placed in a vacuum oven at 34°C for 14 hours to yield Preparation 1.

[0873] Two crystalline solid forms of the GPSSG-NH2 pentamer have been identified, including GPSSG-NH2 Form A and Form B. GPSSG-NH2 Form A is a crystalline solvate that is not physically stable and readily converts to Form B during isolation or upon air drying. GPSSG-NH2 Form B is a crystalline anhydrous / non-solvated form.

[0874] XRPD patterns of crystalline GPSSG-NH2 pentamer were obtained on a Bruker D8 Endeavor X-ray powder diffractometer equipped with a CuKα (1.5418 Å) source and a Linxeye detector, operating at 40 kV and 40 mA. Samples were scanned from 4 to 42° 2θ with a step size of 0.009° 2θ and a scan rate of 0.5 s / step, using a 0.3° primary slit aperture and a 3.9° particle size distribution (PSD) aperture. For GPSSG-NH2 Form A, samples were scanned from 4 to 25° 2θ at a scan rate of 0.1 s / step. Powders were packed into quartz or silicon sample holders, and a smooth surface was obtained using a glass slide. Diffraction patterns of the crystalline forms were collected at ambient temperature and relative humidity. Crystalline peak positions were determined using MDI-Jade v7.9.9. It is well known in the field of crystallography that for any crystalline form, the relative intensities of diffraction peaks may vary due to preferred orientation resulting from factors such as crystalline form and crystal habit. When the effect of preferred orientation is present, peak intensities change, but the characteristic peak positions of a polymorph remain unchanged. See, for example, The United States Pharmacopeia #23, National Formulary #18, pages 1843-1844, 1995. It is also well known in the field of crystallography that for any crystalline form, angular peak positions may vary slightly. For example, peak positions may shift due to variations in the temperature at which the sample is analyzed, sample displacement, or the presence or absence of an internal standard. In this case, a peak position variation of ±0.2 2θ° is estimated to account for these potential variations without precluding unambiguous identification of the indicated crystalline form. Confirmation of a crystalline form can be based on any unique combination of characteristic peaks.

[0875] GPSSG-NH2 form A GPSSG-NH2 Form A was prepared in methanol (MeOH) / tetrahydrofuran (THF). 71.0 mg of amorphous GPSSG-NH2 was dissolved in 0.2 mL MeOH at ambient temperature. 3 × 0.2 mL of THF was added to the solution, and the sample was stirred overnight at ambient conditions to yield a soft white solid that, when analyzed wet, was consistent with GPSSG-NH2 Form A.

[0876] A prepared sample of GPSSG-NH2 Form A is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 16 below, specifically, a peak at 4.3° 2-theta in combination with one or more peaks selected from the group consisting of 5.1, 6.1, 8.0, 10.1, and 18.7° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of GPSSG-NH2 Form A is shown in Figure 5A.

[0877] [Table 18]

[0878] GPSSG-NH2 form B In the above example for GPSSG-NH2 Form A, an additional 3 x 0.2 mL of THF and 1 x 0.2 mL of MeOH were added to the sample. The white slurry was stirred at ambient conditions for 9 days and then isolated by centrifugation using a centrifuge tube filter. The resulting solid was consistent with GPSSG-NH2 Form B and remained Form B after drying under vacuum at 32-33 °C for 1 day.

[0879] A prepared sample of GPSSG-NH2 Form B is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 17 below, specifically peaks at 5.2 and 9.0 degrees 2-theta, in combination with one or more peaks selected from the group consisting of 6.7, 10.0, 10.3, 16.4, 17.8, 18.3, and 19.4 degrees 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of GPSSG-NH2 Form B is shown in Figure 5B.

[0880] [Table 19]

[0881] Example 5 Fmoc-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-OH

[0882] [ka]

[0883] Resin swelling: The resin (0.500 mmol) was charged into a reactor and allowed to swell with DMF (3 x 10 mL x 20 min).

[0884] Washing after Fmoc removal: After deprotection, the resin was washed with DMF (5 x 10 mL x 2 minutes).

[0885] Post-coupling wash: After coupling, the resin was washed with DMF (5 x 10 mL x 2 min).

[0886] Resin washing and drying: After the last coupling or deprotection, the resin was washed with DMF (5 x 10 mL x 2 min), followed by DCM (5 x 10 mL x 2 min) and evacuated to dryness under N2 atmosphere until constant weight.

[0887] [Table 20]

[0888] Two crystalline solid forms of the TFTSD pentamer have been identified, including TFTSD Form A and Form B. TFTSD Form B is a semi-disordered crystalline material observed from a variety of solvent conditions. It represents a family of isostructural solvates. TFTSD Form A is also a semi-disordered crystalline material produced by desolvation or partial desolvation of TFTSD Form B, and therefore represents another family of desolvated or partially desolvated isostructural forms.

[0889] XRPD patterns of crystalline TFTSD pentamer were obtained on a Bruker D8 Endeavor X-ray powder diffractometer equipped with a CuKα (1.5418 Å) source and a Linxeye detector, operating at 40 kV and 40 mA. Samples were scanned from 4 to 30 2θ° using a 0.009 2θ° step size and a 0.25 s / step scan rate, a 0.3° primary slit aperture, and a 3.9° particle size distribution (PSD) aperture. In some cases, scan ranges of 4 and 25 2θ° were used, with a 0.009 2θ° step size and a 0.1 s / step scan rate. Samples were packed into quartz or silicon sample holders, and smooth surfaces were obtained using glass slides. Crystalline form diffraction patterns were collected at ambient temperature and relative humidity. Crystalline peak positions were determined using MDI-Jade v7.9.9. It is well known in the field of crystallography that for any crystalline form, the relative intensities of diffraction peaks may vary due to preferred orientation resulting from factors such as crystalline form and crystal habit. When the effect of preferred orientation is present, peak intensities change, but the characteristic peak positions of a polymorph remain unchanged. See, for example, The United States Pharmacopeia #23, National Formulary #18, pages 1843-1844, 1995. It is also well known in the field of crystallography that for any crystalline form, angular peak positions may vary slightly. For example, peak positions may shift due to variations in the temperature at which the sample is analyzed, sample displacement, or the presence or absence of an internal standard. In this case, a peak position variation of ±0.2 2θ° is estimated to account for these potential variations without precluding unambiguous identification of the indicated crystalline form. Confirmation of a crystalline form can be based on any unique combination of characteristic peaks.

[0890] TFTSD form A In one experiment, 171.5 mg of amorphous TFTSD pentamer was dissolved in 0.7 mL EtOAc at ambient conditions. 0.7 mL heptane was slowly added to the sample while stirring, resulting in immediate precipitation. After adding an additional 0.7 mL heptane, the sample was stirred at ambient conditions for 4 days, resulting in a white slurry. The white solid isolated from the sample was consistent with TFTSD Form A.

[0891] In another experiment, 45.7 mg of amorphous TFTSD pentamer was dissolved in 100 μL MEK to give a clear solution, 200 μL n-propyl ether was added to the solution, and the sample was stirred at ambient conditions for 4 days to give a white slurry. TFTSD Form A was produced by removing a wet aliquot from the slurry and air-drying it at ambient conditions for 1 day.

[0892] A prepared sample of TFTSD Form A is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 18 below, specifically, a peak at 6.7-7.1°2-theta in combination with one or more peaks selected from the group consisting of 5.8-6.1 and 8.8-9.0°2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of TFTSD Form A is shown in Figure 6A.

[0893] [Table 21]

[0894] TFTSD form B In one experiment, 195.8 mg of amorphous TFTSD pentamer was dissolved in 0.4 mL THF at ambient conditions. While stirring, 0.4 mL heptane was slowly added to the sample, resulting in immediate precipitation. The sample was stirred at ambient conditions for 4 days, and a white slurry was observed. The wet solid from the slurry was consistent with TFTSD Form B.

[0895] In another experiment, 45.7 mg of amorphous TFTSD pentamer was dissolved in 100 μL MeOAc to give a clear solution, and 100 μL heptane was added to the solution, immediately forming a white precipitate. The sample was stirred at ambient conditions for 4 days, and a wet sample from the resulting white slurry was consistent with TFTSD Form B.

[0896] A prepared sample of TFTSD Form B is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 19 below, specifically, a peak at 5.0-5.2° 2-theta in combination with one or more peaks selected from the group consisting of 5.3-5.4, 5.7-6.0, 7.6-7.9, and 8.7-9.1° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of TFTSD Form B is shown in Figure 6B.

[0897] [Table 22]

[0898] Example 6: Boc-Tyr(tBu)-Aib-Gln(trt)-Gly-OH

[0899] [ka]

[0900] Conventional SPPS The tetramer is synthesized via traditional SPPS.

[0901] Loading of Fmoc-Gly-OH onto CTC resin The CTC resin was swollen with DMF and loaded with Fmoc-Gly-OH using 1.5 molar equivalents of Gly and 4.0 equivalents of DIEA relative to the loading factor of the CTC resin. The loading was carried out at 25°C for 4 hours. After washing the resin, unreacted sites on the resin were capped with a solution of 0.5 volumes of MeOH, 1.6 volumes of DIEA, and 8 volumes of DMF relative to the weight of the resin used.

[0902] Coupling with Fmoc-L-Gln(trt)-OH The Fmoc group was removed by treatment with 10 volumes of 20% piperidine / DMF for 1 hour, and the resin was washed with isopropyl acetate until residual piperidine was less than 500 ppm. A solution of Fmoc-L-Gln(trt)-OH (2.0 equiv.) and Oxyma (2.0 equiv.) in 5.3 volumes of DMF was prepared. DIC (2.2 equiv.) was added, and the amino acid was preactivated at 20°C for 90 minutes. The activated ester solution was added to the resin, and the reaction mixture was stirred at 20°C for 12 hours, then drained. The resin was washed with isopropyl acetate and carried forward to the next coupling.

[0903] Coupling with Fmoc-Aib-OH The Fmoc group was removed by treatment with 10 volumes of 20% piperidine / DMF for 1 hour, and the resin was washed with isopropyl acetate until residual piperidine was less than 500 ppm. A solution of Fmoc-Aib-OH (2.0 equiv.) and Oxyma (2.0 equiv.) in 5.3 volumes of DMF was prepared. DIC (2.2 equiv.) was added, and the amino acid was preactivated at 20°C for 15 minutes. The activated ester solution was added to the resin, and the reaction mixture was stirred at 20°C for 12 hours, then drained. The resin was washed with isopropyl acetate and carried forward to the next coupling.

[0904] Coupling with Boc-L-Tyr(tBu)-OH The Fmoc group was removed by treatment with 10 volumes of 20% piperidine / DMF for 1 hour, and the resin was washed with isopropyl acetate until residual piperidine was less than 500 ppm. A solution of Boc-L-Tyr(tBu)-OH (2.0 equiv.) and Oxyma (2.0 equiv.) in 5.3 volumes of DMF was prepared. DIC (2.2 equiv.) was added, and the amino acid was preactivated at 20°C for 15 minutes. The activated ester solution was added to the resin, and the reaction mixture was stirred at 20°C for 12 hours and then drained. The resin was washed with isopropyl acetate and dried under vacuum at 35°C to constant weight.

[0905] Disconnect All cleavage procedures were performed at 15-25°C. The tetramer-on-resin material was placed in a cleavage filter reactor, 5 L of DCM / kg resin was added, the material was stirred for at least 10 minutes, and the solvent was drained and discarded. Next, 5 L of 0.5 vol% TFA resin in DCM / kg was added to the cleavage filter reactor, the mixture was stirred for up to 60 minutes, the product solution was drained into a product collection vessel, and neutralized with 1 equivalent of pyridine. Next, 5 L of 0.5 vol% TFA resin in DCM / kg was added to the cleavage filter reactor, the mixture was stirred for up to 60 minutes, the product solution was drained into a product collection vessel, and neutralized with 1 equivalent of pyridine. Next, 5 L of DCM / kg was added to the cleavage filter reactor, which was stirred for up to 10 minutes. The product solution was drained into a product collection vessel, and 5 L of DCM / kg was added to the cleavage filter reactor, which was stirred for up to 10 minutes. The product solution was drained into a product collection vessel.

[0906] Post-processing The product solution was concentrated to 5 L / kg resin under a pressure of 0.4 bar or less and a jacket temperature of 15-25°C. 10 L of purified water / kg resin was added, and the mixture was stirred for ≥30 min and allowed to settle until clear. The bottom organic layer was collected and subjected to Karl Fischer (KF) titration, as well as pyridine and TFA in-process specifications. If the KF value exceeded 0.35 wt.%, 0.4 kg of Na2SO4 / kg resin was added for reslurry drying. If the pyridine value exceeded 0.2 wt.%, the corresponding equivalent of 0.5% TFA in DCM was added to titrate the excess pyridine. The mixture was stirred for ≥10 min, and 10 L of water / kg resin was added. The mixture was stirred for ≥30 min. The organic layer was collected. Once the in-process specifications were met, 2 L of pentyl acetate / kg resin was added, and the mixture was concentrated to 2.5 L of solution / kg resin under a 40°C jacket and 0.1 bar pressure. The solution IPC was checked at less than 7 wt.% DCM and an additional 2 L pentyl acetate / kg resin was added.

[0907] LPPS Alternatively, the tetramer is synthesized via LPPS.

[0908] Step 1:

[0909] [ka]

[0910] Aib protection: Place H-Aib-OH (1.3 equivalents relative to Boc-Tyr(But)-OH) and MeCN (3V) in one vessel, then slowly add bis(trimethylsilyl)acetamide (BSA) (1 equivalent). Stir the mixture at 20-30°C for 16 hours.

[0911] Coupling reaction--In a separate vessel, place Boc-Tyr(But)-OH, MeCN (7V), and 2,6-lutidine (3 eq). Adjust the temperature of the vessel to -25°C. Stir the mixture for 0.5 h. Slowly charge PivCl (1.1 eq) to the vessel. Stir the mixture for 2 h. Analyze a sample for complete activation. Charge the contents of the first vessel (Aib protected) to this reaction vessel. Stir for 16 h at -25°C. Analyze a sample for reaction completion.

[0912] Quench and Workup - Add acetic acid (2 eq) and water (10 V). Concentrate to remove MeCN. Extract twice with EtOAc (10 V). Wash the organic layer with 5% citric acid (10 V) and water (10 V). Concentrate the organic layer to 2 V. Slowly add heptane (16 V) and then filter the solid. Redissolve the solid in EtOAc (6 V) and heat to 40°C. Slowly add heptane (24 V), then slowly cool to 20°C and mix for 16 hours. Filter and dry the wet cake under vacuum at 35-45°C for 24 hours.

[0913] Step 2:

[0914] [ka]

[0915] Coupling reaction—Add a solution of dimeric intermediate (C18070201-B) in THF (3 V) and NMM (1.35 eq.) to a solution of IBCF (1.05 eq.) in THF (7 V) at 0°C. Stir at 0°C for 4-6 h. Check sample for complete activation. In a separate reaction vessel, dissolve H-Gln(Trt)-OH (1.4 eq.) in THF (10 V) and water (3 V), stir for 0.5 h, and then add to the solution containing dimeric intermediate B at 0°C. Add DIPEA (3.0 eq.) and stir at 0°C for 4-6 h. Raise the temperature to 20°C over 3-4 h, then stir for 12-20 h. Check sample for reaction completion. Concentrate under vacuum below 45°C to 1-2 V. Add MTBE (20 V), then wash the organic layer four times with 10:1 5% aqueous KHSO4:DMF (15 V). Wash the organic layer four times with 10:1 5% aqueous Na2CO3:DMF (10:1). Wash the organic layer once with 5% aqueous KHSO4 (10 V). Concentrate to 1-2 V.

[0916] Salt formation purification—Add EtOAc (2.67 V) to the solution and heat to 50° C. Add (1S)-1-phenylpropan-1-amine (1.5 equiv.), then slowly add MeCN (5.33 V). Stir at 50° C. for 2 h, then slowly cool to 25° C. Filter and wash the cake with 1:2 EtOAc:MeCN (1 V). Add MTBE (20 V), then wash twice with 5% KHSO4. Concentrate to dryness and take a sample for analysis. Repeat salt formation purification if necessary.

[0917] Isolation - Add MTBE (1.2 V) to the solid. Add heptane (6 V) and stir at 25° C. for 16 hours. Filter and wash the wet cake with heptane (2 V). Dry the wet cake at 40° C. for 18 hours.

[0918] Step 3:

[0919] [ka]

[0920] Add the trimer intermediate (C18070201-EA), H-Gly-OBzl (1.3 equiv.), and MeCN (10 V) to the reaction vessel and reduce the temperature to -20 °C. Add 2,6-lutidine (3.0 equiv.) and COMU (1.3 equiv.) and stir at -20 °C for 2-4 h. Analyze a sample to test for reaction completion. Concentrate to 1 V below 46 °C. Add EtOAc (5 V) and stir for 0.5-1 h. Add MTBE (5 V), then filter the mixture. Wash the cake with 1:1 EtOAc:MTBE (5 V). Wash the filtrate four times with 5% NaHCO3 (10 V), four times with KHSO4 (10 V), and then once with water (10 V). Concentrate the organic layer to 1-2 V below 45 °C. Add DMF (2.5 V) and stir to form a clear solution at 15-20°C. Slowly add water (7.5 V). Stir the mixture at 15-20°C for 2-6 hours. Filter and wash the cake with water (2-3 V). Reslurry the cake in water (10 V) for 2-6 hours at 15-20°C and filter. Wash the cake with water (2-3 V) and then dry at 45°C for 16-48 hours. Check for residual DMF and water. Re-slurry again in water or dry further if necessary.

[0921] Step 4:

[0922] [ka]

[0923] Benzyl deprotection—tetrameric ester intermediate (C18070201-CA) and IPA (10 V) are added to the reaction vessel. Pd / C (0.1×, 50 wt%) is added to the reaction vessel and the atmosphere is exchanged with argon three times. The atmosphere is exchanged with hydrogen three times. The vessel pressure is adjusted to 45 psi with hydrogen and the reactor is heated to 40°C. The reaction is stirred at 45 psi and 40°C for 16–20 hours. A sample is taken for analysis. The reaction vessel is cooled to 20–30°C and the atmosphere is exchanged with argon three times. The mixture is filtered and the wet cake (Pd / C) is washed with IPA (2 V). The filtrate is concentrated to 1 V below 45°C.

[0924] Crystallization - Charge pentyl acetate (5) and then concentrate to 1V below 45°C. Repeat this process a total of three times. Charge pentyl acetate (5V) and stir at 20°C for 0.5-1 hour. Charge heptane (1V) over 1 hour. Stir the mixture at 20°C for 3-5 hours. Apply two thermal cycles (heat to 35°C over 1 hour, stir for 3-5 hours, cool to 20°C over 1 hour, stir at 20°C for 3-5 hours). Charge 5V heptane over 1 hour. Stir at 20°C for 3-5 hours. Filter the solids and wash the wet cake with heptane (2V). Dry the wet cake under vacuum at 40-50°C for 16-24 hours. Take a sample to determine if reslurrying is necessary.

[0925] Crystallization After solvent exchange by distillation, the tetramer was in 5 volumes of pentyl acetate (L / kg resin, tetramer concentration 80-100 mg / mL). For primary nucleation, the solution was held at 20 °C for 12 hours. If solids were observed, the mixture was advanced to thermal cycling. If no solids were observed, the mixture was cooled to 5 °C and held for 12 hours. If solids were observed, the mixture was advanced to thermal cycling. If no solids were observed, 1 volume of heptane was added and the mixture was held for 12 hours. If solids were observed, the mixture was advanced to thermal cycling. If no solids were observed, 1 volume of heptane was added and the process was repeated.

[0926] A thermal cycle was used to convert the amorphous form to a crystalline form and grow the crystal particles. The mixture was heated to 30°C and held for 1 hour, then cooled to 20°C and held for 1 hour. The thermal cycle was repeated five times.

[0927] After thermal cycling, 5 volumes of heptane were added to reduce solubility and improve yield. The slurry was then filtered. It was washed once with 5 volumes of 1:1 heptane:pentyl acetate to remove impurities. It was washed twice with 5 volumes of heptane in a reslurry wash to remove pentyl acetate. Finally, the solid was dried at 50°C.

[0928] Two solid forms of the Y-Aib-QG tetramer have been identified, including Y-Aib-QG Form A and Form B. Y-Aib-QG Form A is a crystalline form. Solvated forms were initially generated from pentyl acetate or mixtures containing pentyl acetate (e.g., pentyl acetate / ETBE, pentyl acetate / TAME, pentyl acetate / heptane).

[0929] Y-Aib-QG Form A represents a family of isostructural solvates and can also be generated from other solvent conditions such as EtOAc and 2-Me THF / TAME.

[0930] Y-Aib-QG Form B is a semi-disordered crystalline material produced by desolvation or partial desolvation of Y-Aib-QG Form A and therefore represents another family of desolvated or partially desolvated isostructural forms.

[0931] Preparation of Y-Aib-QG Form A Y-Aib-QG Form A was obtained by adding 1 mL of pentyl acetate to 210.0 mg of amorphous Y-Aib-QG tetramer. The sample was stirred at ambient conditions to obtain a clear solution. 3 mL of ETBE (t-butyl ethyl ether) was added to the solution, and the sample was capped and stirred at ambient conditions. A thick white slurry was obtained, and the sample was stirred for 2 days, after which an additional 2 mL of ETBE was added to the slurry. The sample was stirred for an additional 3 days at ambient conditions, and a wet sample from the resulting white slurry was consistent with Y-Aib-QG Form A.

[0932] Y-Aib-QG Form A was also obtained by dissolving 55.5 mg of amorphous Y-Aib-QG tetramer in 250 μL of pentyl acetate. 0.5 mL of heptane was added to the solution, resulting in a white suspension. The sample was stirred in a capped vial at ambient conditions for 1 day, resulting in a white slurry. XRPD of a wet sample from the slurry was consistent with Y-Aib-QG Form A.

[0933] Preparation of Y-Aib-QG Form B Form B solid was obtained from drying Y-Aib-QG Form A. The wet solid in the first example above was isolated by vacuum filtration, and the resulting material was rinsed on the filter with 0.5 mL of ETBE, air-dried on the filter under continuous vacuum for about 5 minutes, collected, and then dried under vacuum at about 30° C. for 1 day. XRPD of the resulting white solid was consistent with Y-Aib-QG Form B.

[0934] The XRPD pattern of the crystalline Y-Aib-QG tetramer was obtained on a Bruker D8 Endeavor X-ray powder diffractometer equipped with a CuKα (1.5418 Å) source and a Linxeye detector, operating at 40 kV and 40 mA. The sample was scanned from 4 to 30° 2θ with a step size of 0.009° 2θ and a scan rate of 0.25 s / step, using a 0.3° primary slit aperture and a 3.9° particle size distribution (PSD) aperture. The powder was packed into a quartz or silicon sample holder, and a smooth surface was obtained using a glass slide. The crystalline form diffraction pattern was collected at ambient temperature and relative humidity. Crystalline peak positions were determined using MDI-Jade v7.9.9. It is well known in the field of crystallography that for any crystalline form, the relative intensities of diffraction peaks may vary due to preferred orientation resulting from factors such as crystalline morphology and crystal habit. When preferred orientation effects exist, peak intensities change, but the characteristic peak positions of a polymorph remain unchanged. See, for example, The United States Pharmacopeia #23, National Formulary #18, pages 1843-1844, 1995. Furthermore, it is well known in the field of crystallography that angular peak positions may vary slightly for any crystalline form. For example, peak positions may shift due to variations in the temperature at which the sample is analyzed, sample displacement, or the presence or absence of an internal standard. In this case, a peak position variation of ±0.2 2θ° is estimated to account for these potential variations without precluding unambiguous identification of the indicated crystalline form. Confirmation of a crystalline form can be based on any unique combination of characteristic peaks.

[0935] XRPD of Y-Aib-QG Form A The prepared sample of Y-Aib-QG Form A is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 20 below, specifically, a peak at 6.3-6.4° 2-theta in combination with one or more peaks selected from the group consisting of 4.5, 7.1, 13.0-13.1, 15.9-16.0, and 18.4-18.6° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of Y-Aib-QG Form A is shown in Figure 7A.

[0936] [Table 23]

[0937] XRPD of Y-Aib-QG Form B The prepared sample of Y-Aib-QG Form B is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 21 below, specifically, a peak at 7.0-7.2° 2-theta in combination with one or more peaks selected from the group consisting of 5.0-5.4, 7.6-7.7, 8.8-8.9, 9.4-9.5, and 12.5-12.7° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of Y-Aib-QG Form B is shown in Figure 7B.

[0938] [Table 24]

[0939] Example 7: (Boc)-His(dnp)-Aib-Gln(trt)-Gly

[0940] [ka]

[0941] Three crystalline solid forms of the H(DNP)-Aib-QG tetramer have been identified, including H(DNP)-Aib-QG Form A, Form B, and Form C.

[0942] H(DNP)-Aib-QG Form A is a crystalline unstable solvate that is readily converted (during isolation or upon drying) to H(DNP)-Aib-QG Form C.

[0943] H(DNP)-Aib-QG Form B is a crystalline material produced from multiple solvent conditions. It represents a family of isostructural solvates.

[0944] H(DNP)-Aib-QG Form C is a semi-disordered crystalline material observed from multiple solvent conditions.

[0945] H(DNP)-Aib-QG Form A H(DNP)-Aib-QG Form A was prepared from acetonitrile (ACN) / methyl tert-butyl ether (MTBE). Approximately 50 mg of amorphous H(DNP)-Aib-QG tetramer was dissolved in 0.2 mL of 1:5 (vol / vol) ACN / MTBE at ambient conditions to give a clear yellow solution. With stirring, 0.1 mL of MTBE was added to the solution to give a pale yellow slurry. The sample was stirred at ambient conditions for 5 days, and the wet solid was consistent with H(DNP)-Aib-QG Form A.

[0946] H(DNP)-Aib-QG form B Approximately 50 mg of amorphous H(DNP)-Aib-QG tetramer was dissolved in 0.2 mL of methyl acetate (MeOAc) at ambient conditions to give a clear, yellow solution. Five 0.2 mL portions of cyclopentyl methyl ether (CPME) were added to the sample. The solution was stirred overnight at ambient conditions and then placed in a refrigerator at 2-8 °C for approximately 6 weeks. A white solid emerged from the solution, consistent with H(DNP)-Aib-QG Form B.

[0947] Following a similar procedure, H(DNP)-Aib-QG form B was also produced from a 1:10 (vol / vol) nitromethane / MTBE solution at 2–8 °C.

[0948] Approximately 50 mg of amorphous H(DNP)-Aib-QG tetramer was dissolved in 0.2 mL of ethyl acetate (EtOAc) at ambient conditions to give a clear, yellow solution. With stirring, 0.2 mL of heptane was added to the solution, producing a yellow, sticky substance. The sample was stirred at 36°C for 7 days to give a pale yellow slurry. The resulting solid was consistent with H(DNP)-Aib-QG Form B.

[0949] H(DNP)-Aib-QG Form C H(DNP)-Aib-QG Form C was prepared from tetrahydrofuran (THF) / MTBE. Approximately 50 mg of amorphous H(DNP)-Aib-QG tetramer was dissolved in 0.1 mL of THF at ambient conditions to give a clear, yellow solution. With stirring, 5 × 0.1 mL of MTBE was added, and the sample was stirred overnight at ambient conditions and then at 36 °C for 7 days, resulting in a pale yellow slurry. The sample was isolated by centrifugation at ambient conditions using a centrifuge tube filter. The resulting solid was consistent with H(DNP)-Aib-QG Form C.

[0950] Following a similar procedure, H(DNP)-Aib-QG Form C can be produced from different solvent systems containing MTBE, such as EtOAc / MTBE, methyl ethyl ketone (MEK) / MTBE.

[0951] XRPD patterns of the crystalline H(DNP)-Aib-QG tetramer were obtained on a Bruker D8 Endeavor X-ray powder diffractometer equipped with a CuKα (1.5418 Å) source and a Linxeye detector, operating at 40 kV and 40 mA. Samples were scanned from 4 to 42° 2θ with a step size of 0.009° 2θ and a scan rate of 0.5 s / step, using a 0.3° primary slit aperture and a 3.9° particle size distribution (PSD) aperture. For Form A, samples were scanned from 4 to 25° 2θ at a scan rate of 0.1 s / step. Powders were packed into quartz or silicon sample holders, and a smooth surface was obtained using a glass slide. Diffraction patterns of the crystalline forms were collected at ambient temperature and relative humidity. Crystalline peak positions were determined using MDI-Jade v7.9.9. It is well known in the field of crystallography that for any crystalline form, the relative intensities of diffraction peaks may vary due to preferred orientation resulting from factors such as crystalline form and crystal habit. When the effect of preferred orientation is present, peak intensities change, but the characteristic peak positions of a polymorph remain unchanged. See, for example, The United States Pharmacopeia #23, National Formulary #18, pages 1843-1844, 1995. It is also well known in the field of crystallography that for any crystalline form, angular peak positions may vary slightly. For example, peak positions may shift due to variations in the temperature at which the sample is analyzed, sample displacement, or the presence or absence of an internal standard. In this case, a peak position variation of ±0.2 2θ° is estimated to account for these potential variations without precluding unambiguous identification of the indicated crystalline form. Confirmation of a crystalline form can be based on any unique combination of characteristic peaks.

[0952] XRPD of H(DNP)-Aib-QG Form A The prepared sample of H(DNP)-Aib-QG Form A is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 22 below, specifically, a peak at 4.8° 2-theta in combination with one or more peaks selected from the group consisting of 5.6, 6.2, 14.8, and 15.6° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of H(dnp)-Aib-QG Form A is shown in Figure 8A.

[0953] [Table 25]

[0954] XRPD of H(DNP)-Aib-QG Form B The prepared sample of H(DNP)-Aib-QG Form B is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 23 below, specifically, a peak at 5.3° 2-theta in combination with one or more peaks selected from the group consisting of 7.7, 10.5, 11.3, 11.6, and 14.4° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of H(dnp)-Aib-QG Form B is shown in Figure 8B.

[0955] [Table 26]

[0956] XRPD of H(DNP)-Aib-QG Form C The prepared sample of H(DNP)-Aib-QG Form C is characterized by an XRPD pattern using CuKα radiation, with diffraction peaks at 6.2 and 6.9° 2-theta (2-theta values) with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of H(dnp)-Aib-QG Form C is shown in Figure 8C.

[0957] Example 8: (Boc)His(trt)-Aib-Gln(trt)-Gly

[0958] [ka]

[0959] Two crystalline solid forms of the H(trt)-Aib-QG tetramer have been identified, including H(trt)-Aib-QG Form A and Form B.

[0960] H(trt)-Aib-QG Form A is a crystalline material produced from multiple organic mixtures. It represents a family of isostructural solvates with varying amounts of solvent content in the unit cell.

[0961] H(trt)-Aib-QG Form B is a crystalline material prepared from EtOH / HO. It is a solvated form.

[0962] H(trt)-Aib-QG form A H(trt)-Aib-QG Form A was prepared from tetrahydrofuran (THF) / methyl tert-butyl ether (MTBE). 50.3 mg of amorphous H(trt)-Aib-QG tetramer was stirred in 0.2 mL of 1:3 (vol / vol) THF / MTBE at ambient conditions for 5 days to give a thick white slurry. 0.2 mL of 1:3 (vol / vol) THF / MTBE was added, and the sample was stirred at ambient conditions for an additional 3 days before isolation. The white solid from the slurry, either wet or isolated, was consistent with H(trt)-Aib-QG Form A.

[0963] Following a similar procedure, H(trt)-Aib-QG Form A can also be prepared from other organic solvent mixtures such as THF / heptane and 1,4-dioxane / HO, acetonitrile (ACN) / MTBE, and ethyl acetate / MTBE.

[0964] H(trt)-Aib-QG form B 50.3 mg of amorphous H(trt)-Aib-QG tetramer was stirred in 0.2 mL of 1:1 (vol / vol) ethanol / HO at ambient conditions, resulting in a mixture of white particles and a pale yellow gel after 5 days. A 50 μL aliquot from this slurry was added as a seed to a sample containing 311.2 mg of amorphous H(trt)-Aib-QG tetramer in 1 mL of 1:1 (vol / vol) ethanol / HO. The sample was stirred at ambient conditions for 5 days. The wet solid isolated from the sample was consistent with H(trt)-Aib-QG Form B.

[0965] XRPD patterns of the crystalline H(trt)-Aib-QG tetramer were obtained on a Bruker D8 Endeavor X-ray powder diffractometer equipped with a CuKα (1.5418 Å) source and a Linxeye detector, operating at 40 kV and 40 mA. Samples were scanned from 4 to 42 2θ° with a step size of 0.009 2θ° and a scan rate of 0.5 s / step, using a 0.3° primary slit aperture and a 3.9° particle size distribution (PSD) aperture. In some cases, samples were scanned from 4 to 30 2θ° at a scan rate of 0.25 s / step. Powders were packed into quartz or silicon sample holders, and smooth surfaces were obtained using glass slides. Crystalline form diffraction patterns were collected at ambient temperature and relative humidity. Crystalline peak positions were determined using MDI-Jade v7.9.9. It is well known in the field of crystallography that for any crystalline form, the relative intensities of diffraction peaks may vary due to preferred orientation resulting from factors such as crystalline form and crystal habit. When the effect of preferred orientation is present, peak intensities change, but the characteristic peak positions of a polymorph remain unchanged. See, for example, The United States Pharmacopeia #23, National Formulary #18, pages 1843-1844, 1995. It is also well known in the field of crystallography that for any crystalline form, angular peak positions may vary slightly. For example, peak positions may shift due to variations in the temperature at which the sample is analyzed, sample displacement, or the presence or absence of an internal standard. In this case, a peak position variation of ±0.2 2θ° is estimated to account for these potential variations without precluding unambiguous identification of the indicated crystalline form. Confirmation of a crystalline form can be based on any unique combination of characteristic peaks.

[0966] XRPD of H(trt)-Aib-QG Form A A prepared sample of H(trt)-Aib-QG Form A is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 24 below, specifically, a peak at 4.7° 2-theta in combination with one or more peaks selected from the group consisting of 5.5, 8.2, 10.1, 11.8, 13.3, 13.6, and 18.9° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of H(trt)-Aib-QG Form A is shown in Figure 9A.

[0967] [Table 27]

[0968] XRPD of H(trt)-Aib-QG Form B A prepared sample of H(trt)-Aib-QG Form B is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 25 below, specifically, a peak at 5.8° 2-theta in combination with one or more peaks selected from the group consisting of 5.3, 8.9, 9.2, 15.2, 18.6, and 19.5° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of H(trt)-Aib-QG Form B is shown in Figure 9B.

[0969] [Table 28]

[0970] Example 9: (Fmoc)Asp(t-Bu)-Tyr(t-Bu)-Ser(t-Bu)-Lys

[0971] [ka]

[0972] Three crystalline solid forms of the OXM DYSK tetramer have been identified, including DYSK forms A, B, and C.

[0973] DYSK Form A is a crystalline material produced from multiple organic mixtures. It is a solvated form that exists in wet conditions.

[0974] DYSK Form B is a crystalline material that is anhydrous / non-solvated.

[0975] DYSK Form C is a semi-disordered crystalline material produced from an MTBE / ETBE mixture. It is a solvated material.

[0976] DYSK form A DYSK Form A was prepared from methyl acetate (MeOAc) / dibutyl ether. 0.2 mL of MeOAc was added to 40.8 mg of amorphous DYSK tetramer to obtain a clear solution, and 3 × 0.2 mL of dibutyl ether was added to the solution. After overnight slurrying at ambient temperature, a white gel formed. The gel was broken down into a viscous liquid and stirred at ambient conditions for 1 day, then at approximately 36 °C for 5 days to obtain a thick white slurry. The wet solid from the slurry was consistent with DYSK Form A.

[0977] Following similar procedures, DYSK Form A can also be prepared from other organic solvent mixtures, such as acetone / dibutyl ether, acetonitrile (ACN) / dibutyl ether, ethyl acetate (EtOAc) / dibutyl ether, MeOAc / heptane, and methyl ethyl ketone (MEK) / dibutyl ether.

[0978] DYSK form B DYSK Form B was prepared from methyl acetate (MeOAc) / ethyl tert-butyl ether (ETBE). 0.2 mL of MeOAc was added to 40.8 mg of amorphous DYSK tetramer to obtain a clear solution, and 3 × 0.2 mL of ETBE was added to the solution, followed by stirring at ambient conditions for a total of 7 days. During stirring, a white gel was observed, which slowly transformed into a thick white slurry. The solids from the slurry were either wet or isolated, consistent with DYSK Form B.

[0979] Following similar procedures, DYSK Form B can also be prepared from other organic solvent mixtures such as ethyl acetate (EtOAc) / ETBE, methyl ethyl ketone (MEK) / ETBE, MEK / heptane, and pentyl acetate / ETBE.

[0980] DYSK Form B was also observed from DYSK Form A slurries when the solids were isolated using a centrifuge tube filter and subsequently air-dried at ambient conditions for 4 hours or more.

[0981] DYSK form C 40.4 mg of amorphous DYSK solid was dissolved in 0.2 mL of MTBE by briefly heating the mixture at 62 °C. 3 × 0.2 mL of ETBE was added to the clear solution at ambient conditions, and after overnight slurrying at ambient conditions, a white gel formed. The gel broke into a viscous liquid and was stirred at ambient conditions for an additional 6 days to yield a thick white slurry. The wet solid from the slurry was consistent with DYSK Form C.

[0982] XRPD patterns of the crystalline TFTS tetramer were obtained on a Bruker D8 Endeavor X-ray powder diffractometer equipped with a CuKα (1.5418 Å) source and a Linxeye detector, operating at 40 kV and 40 mA. Samples were scanned from 4 to 42° 2θ with a step size of 0.009° 2θ and a scan rate of 0.5 s / step, using a 0.3° primary slit aperture and a 3.9° particle size distribution (PSD) aperture. For Forms A and C, samples were scanned from 4 to 25° 2θ at a scan rate of 0.1 s / step. Powders were packed into quartz or silicon sample holders, and a smooth surface was obtained using a glass slide. Diffraction patterns of the crystalline forms were collected at ambient temperature and relative humidity. Crystalline peak positions were determined using MDI-Jade v7.9.9. It is well known in the field of crystallography that for any crystalline form, the relative intensities of diffraction peaks may vary due to preferred orientation resulting from factors such as crystalline form and crystal habit. When the effect of preferred orientation is present, peak intensities change, but the characteristic peak positions of a polymorph remain unchanged. See, for example, The United States Pharmacopeia #23, National Formulary #18, pages 1843-1844, 1995. It is also well known in the field of crystallography that for any crystalline form, angular peak positions may vary slightly. For example, peak positions may shift due to variations in the temperature at which the sample is analyzed, sample displacement, or the presence or absence of an internal standard. In this case, a peak position variation of ±0.2 2θ° is estimated to account for these potential variations without precluding unambiguous identification of the indicated crystalline form. Confirmation of a crystalline form can be based on any unique combination of characteristic peaks.

[0983] XRPD of DYSK Form A A prepared sample of DYSK Form A is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth below in Table 26, specifically a peak at 5.3° 2-theta in combination with one or more peaks selected from the group consisting of 6.0, 6.9, 7.2, 8.0, 12.2, and 15.6° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of DYSK Form A is shown in Figure 10A.

[0984] [Table 29]

[0985] XRPD of DYSK Form B A prepared sample of DYSK form B is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth below in Table 27, specifically a peak at 5.8° 2-theta in combination with one or more peaks selected from the group consisting of 4.4, 6.6, 10.1, 11.4, 13.4, and 15.5° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of DYSK form B is shown in Figure 10B.

[0986] [Table 30]

[0987] XRPD of DYSK Form C A prepared sample of DYSK form C is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth below in Table 28, specifically peaks at 4.5 and 5.5 degrees 2-theta in combination with one or more peaks selected from the group consisting of 6.0 and 7.3 degrees 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of DYSK form C is shown in Figure 10C.

[0988] [Table 31]

[0989] Example 10: (Fmoc)Tyr-Ser(t-Bu)-Lys(Boc)-Tyr(t-Bu)

[0990] [ka]

[0991] Multiple crystalline solid forms of the OXM YSKY tetramer have been identified, including YSKY Forms A through E. A polymorphic map illustrating their relationship is shown in Figure 11. They are desolvated solvates generated from solvated forms generated from EtOH (Form B) or IPA (Form C). Form A converts to Form D upon heating to 150°C. Form D has also been observed from MEK.

[0992] YSKY form A 50.3 mg of YSKY amorphous solid was dissolved in 0.8 mL of EtOH at 50° C. The solution was then removed from the 50° C. heating plate and placed in a refrigerator. A solid was observed the next day, which was isolated and air-dried at ambient conditions. The resulting white solid is consistent with YSKY Form A.

[0993] 50.4 mg of YSKY amorphous solid was dissolved in 0.75 mL of IPA at 50° C. The solution was then removed from the 50° C. heating plate and placed in a refrigerator. A solid was observed the next day, which was isolated and air-dried at ambient conditions. The resulting white solid is consistent with YSKY Form A.

[0994] YSKY form B 297.0 mg of YSKY amorphous solid was dissolved in 2.5 mL of EtOH at 54° C. to give a slightly cloudy solution. With stirring, the solution was slowly cooled to ambient temperature on a heating plate with the heater turned off. A white suspension was obtained, and the solid, when analyzed in the wet state, was consistent with YSKY Form B.

[0995] YSKY form C 50.4 mg of YSKY amorphous solid was stirred in 1 mL of IPA overnight at ambient conditions to obtain a clear solution. The solution was allowed to evaporate at ambient conditions from an uncapped vial covered with perforated aluminum foil. The resulting wet solid was consistent with YSKY Form C.

[0996] 223.6 mg of YSKY amorphous solid was dissolved in 1 mL of IPA at 54° C. to give a slightly cloudy solution. While stirring, the solution was slowly cooled to ambient temperature on a heating plate with the heater turned off. A white solid was obtained in solution, which, when analyzed in the wet state, was consistent with YSKY Form C.

[0997] YSKY form D 11.1 mg of YSKY Form A solid was heated to 150°C in a TGA and maintained at that temperature for 5 minutes. The sample was then removed from the TGA instrument and analyzed by XRPD at ambient conditions. The resulting sample was a white solid, consistent with YSKY Form D.

[0998] 50.4 mg of YSKY amorphous solid was dissolved in 0.5 mL of dioxane at ambient temperature to give a clear solution. While stirring, 2 × 0.25 mL of water was added to the solution, resulting in a gel. The sample was vortexed to break up the gel, and the sample was stirred at ambient conditions for 1 day. An aliquot was removed from the white slurry and placed in a clean silicon XRPD sample holder. The sample was allowed to air dry overnight at ambient conditions. The resulting solid was consistent with YSKY Form D.

[0999] XRPD patterns of the crystalline YSKY tetramer were obtained on a Bruker D8 Endeavor X-ray powder diffractometer equipped with a CuKα (1.5418 Å) source and a Linxeye detector, operating at 40 kV and 40 mA. Samples were scanned from 4 to 42° 2θ with a step size of 0.009° 2θ and a scan rate of 0.5 s / step, using a 0.3° primary slit aperture and a 3.9° particle size distribution (PSD) aperture. Samples could also be scanned between 4 and 30° 2θ at a scan rate of 0.25 s / step or between 4 and 25° 2θ at a scan rate of 0.1 s / step. The powder was packed into a silicon sample holder, and a smooth surface was obtained using a glass slide. Crystalline form diffraction patterns were collected at ambient temperature and relative humidity. Crystalline peak positions were determined using MDI-Jade v7.9.9. It is well known in the field of crystallography that for any crystalline form, the relative intensities of diffraction peaks may vary due to preferred orientation resulting from factors such as crystalline form and crystal habit. When the effect of preferred orientation is present, peak intensities change, but the characteristic peak positions of a polymorph remain unchanged. See, for example, The United States Pharmacopeia #23, National Formulary #18, pages 1843-1844, 1995. It is also well known in the field of crystallography that for any crystalline form, angular peak positions may vary slightly. For example, peak positions may shift due to variations in the temperature at which the sample is analyzed, sample displacement, or the presence or absence of an internal standard. In this case, a peak position variation of ±0.2 2θ° is estimated to account for these potential variations without precluding unambiguous identification of the indicated crystalline form. Confirmation of a crystalline form can be based on any unique combination of characteristic peaks.

[1000] XRPD of YSKY Form A A prepared sample of YSKY Form A is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 29 below, specifically peaks at 18.1 and 18.7 degrees 2-theta, in combination with one or more peaks selected from the group consisting of 5.7, 8.7, 13.7, 14.3, 15.9, and 16.2 degrees 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of YSKY Form A is shown in Figure 12A.

[1001] [Table 32]

[1002] XRPD of YSKY Form B A prepared sample of YSKY form B is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 30 below, specifically peaks at 5.9 and 10.5 degrees 2-theta, in combination with one or more peaks selected from the group consisting of 7.1, 8.9, 14.6, and 16.6 degrees 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of YSKY form B is shown in Figure 12B.

[1003] [Table 33]

[1004] XRPD of YSKY Form C A prepared sample of YSKY form C is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 31 below, specifically peaks at 7.8 and 20.3 degrees 2-theta, in combination with one or more peaks selected from the group consisting of 5.8, 15.5, and 19.5 degrees 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of YSKY form C is shown in Figure 12C.

[1005] [Table 34]

[1006] XRPD of YSKY Form D A prepared sample of YSKY form D is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 32 below, specifically peaks at 5.9 and 7.4 degrees 2-theta in combination with one or more peaks selected from the group consisting of 6.5, 6.9, and 14.8 degrees 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of YSKY form D is shown in Figure 12D.

[1007] [Table 35]

[1008] Example 11: Fmoc-GPSSG-NH2 pentamer

[1009] [ka]

[1010] Fmoc-GPSSG-NH2 crystalline form Multiple crystalline solid forms of the OXM Fmoc-protected GPSSG-NH2 pentamer have been identified, including Fmoc-GPSSG-NH2 Forms A to E. A polymorphic map illustrating their relationships is shown in Figure 13. Of these, Form B is a variable solvate formed from acetone, Form A is a desolvated solvate formed from acetonitrile, and Form D is a desolvated solvate formed from MEK.

[1011] Fmoc-GPSSG-NH2 Form A and Form C Fmoc-GPSSG-NH2 Form C was prepared from acetonitrile (ACN). 159.9 mg of solid amorphous Fmoc-GPSSG-NH2 was stirred in 0.5 mL of ACN at ambient conditions, resulting in gel formation. An additional 0.5 mL of ACN was added to the sample to facilitate stirring, and a white slurry was observed the next day. An additional 0.8 mL of ACN was added, and the sample was stirred at ambient conditions for a total of 3 weeks. The resulting white solid, when analyzed wet, was consistent with Fmoc-GPSSG-NH2 Form C.

[1012] The white slurry was transferred to a 0.45 μm nylon centrifuge filter and centrifuged at ambient temperature for 5 minutes to separate the solid from the liquid. The isolated solid was dried overnight in a vacuum oven at ambient temperature. The resulting white solid was consistent with Fmoc-GPSSG-NH2 Form A.

[1013] Fmoc-GPSSG-NH2 Form B Fmoc-GPSSG-NH2 Form B was prepared from acetone. 150.1 mg of solid amorphous Fmoc-GPSSG-NH2 was stirred in 0.5 mL acetone at ambient conditions, resulting in gel formation. An additional 0.5 mL of acetone was added to the sample to facilitate stirring, and a white slurry was observed the next day. An additional 0.3 mL of acetone was added, and the sample was stirred at ambient conditions for a total of 3 weeks. The resulting white solid, either wet or dry, is consistent with Fmoc-GPSSG-NH2 Form B.

[1014] Fmoc-GPSSG-NH2 Form D and Form E Fmoc-GPSSG-NH2 Form E was prepared from methyl ethyl ketone (MEK). 110.4 mg of solid amorphous Fmoc-GPSSG-NH2 was stirred in 0.5 mL of MEK at ambient conditions, resulting in gel formation. An additional 2 × 0.5 mL of MEK was added to the sample to facilitate stirring, and the next day, a mixture of white solid and gel was produced in the sample. The sample was sonicated in a water bath and manually stirred with a spatula to break up the gel, resulting in a white slurry. The sample was stirred at ambient conditions for a total of 3 weeks. The resulting white solid, when analyzed wet, was consistent with Fmoc-GPSSG-NH2 Form E.

[1015] The white slurry was transferred to a 0.45 μm nylon centrifuge filter and centrifuged at ambient temperature for 5 minutes to separate the solid from the liquid. The isolated solid was dried overnight in a vacuum oven at ambient temperature. The resulting white solid was consistent with Fmoc-GPSSG-NH2 Form D.

[1016] XRPD patterns of crystalline Fmoc-GPSSG-NH2 pentamer were obtained on a Bruker D8 Endeavor X-ray powder diffractometer equipped with a CuKα (1.5418 Å) source and a Linxeye detector, operating at 40 kV and 40 mA. Samples were scanned from 4 to 42 2θ° with a step size of 0.009 2θ° and a scan rate of 0.5 s / step, using a 0.3° primary slit aperture and a 3.9° particle size distribution (PSD) aperture. For Fmoc-GPSSG-NH2 Forms C and E, samples were scanned from 4 to 25 2θ° at a scan rate of 0.1 s / step. Powders were packed into quartz or silicon sample holders, and a smooth surface was obtained using a glass slide. Diffraction patterns of the crystalline forms were collected at ambient temperature and relative humidity. Crystalline peak positions were determined using MDI-Jade v7.9.9. It is well known in the field of crystallography that for any crystalline form, the relative intensities of diffraction peaks may vary due to preferred orientation resulting from factors such as crystalline form and crystal habit. When the effect of preferred orientation is present, peak intensities change, but the characteristic peak positions of a polymorph remain unchanged. See, for example, The United States Pharmacopeia #23, National Formulary #18, pages 1843-1844, 1995. It is also well known in the field of crystallography that for any crystalline form, angular peak positions may vary slightly. For example, peak positions may shift due to variations in the temperature at which the sample is analyzed, sample displacement, or the presence or absence of an internal standard. In this case, a peak position variation of ±0.2 2θ° is estimated to account for these potential variations without precluding unambiguous identification of the indicated crystalline form. Confirmation of a crystalline form can be based on any unique combination of characteristic peaks.

[1017] XRPD of Fmoc-GPSSG-NH2 Form A A prepared sample of Fmoc-GPSSG-NH2 Form A is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 33 below, specifically peaks at 5.8 and 18.5 degrees 2-theta, in combination with one or more peaks selected from the group consisting of 8.6, 9.4, 12.9, 13.8, 17.2, and 19.4 degrees 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of Fmoc-GPSSG-NH2 Form A is shown in Figure 14A.

[1018] [Table 36]

[1019] XRPD of Fmoc-GPSSG-NH2 Form B A prepared sample of Fmoc-GPSSG-NH2 Form B is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 34 below, specifically peaks at 7.0-7.1 and 7.5-7.7 degrees 2-theta, in combination with one or more peaks selected from the group consisting of 5.3-5.4, 9.7-9.9, and 14.7-14.9 degrees 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of Fmoc-GPSSG-NH2 Form B is shown in Figure 14B.

[1020] [Table 37]

[1021] XRPD of Fmoc-GPSSG-NH2 Form C A prepared sample of Fmoc-GPSSG-NH2 form C is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 35 below, specifically, a peak at 8.3° 2-theta in combination with one or more peaks selected from the group consisting of 6.3, 11.4, 14.3, and 16.6° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of Fmoc-GPSSG-NH2 form C is shown in Figure 14C.

[1022] [Table 38]

[1023] XRPD of Fmoc-GPSSG-NH2 Form D A prepared sample of Fmoc-GPSSG-NH2 form D is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 36 below, specifically, a peak at 7.2° 2-theta in combination with one or more peaks selected from the group consisting of 6.8, 8.6, 15.8, and 18.9° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of Fmoc-GPSSG-NH2 form D is shown in Figure 14D.

[1024] [Table 39]

[1025] XRPD of Fmoc-GPSSG-NH2 Form E A prepared sample of Fmoc-GPSSG-NH2 form E is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 37 below, specifically, a peak at 6.1° 2-theta in combination with one or more peaks selected from the group consisting of 6.3, 7.8, 10.0, and 12.4° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of Fmoc-GPSSG-NH2 form E is shown in Figure 14E.

[1026] [Table 40]

[1027] Example 12: Fmoc-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-OH

[1028] [ka]

[1029] Crystallization screening was performed on the free form of Fmoc-GPSSG-OH using a variety of solvents and solvent mixtures. Different crystallization methods were used, including solvent-based techniques such as slurry, cooling, ambient and sub-ambient temperature holds, solvent / anti-solvent addition, or a combination of techniques, and non-solvent-based techniques such as thermal stress.

[1030] Two solid forms were identified: Fmoc-GPSSG-OH Form A and Fmoc-GPSSG-OH Form B. Form A was a solvated form produced from 1-propanol (1-PrOH) or mixtures containing 1-PrOH (such as 1-PrOH / heptane). Form B was a desolvated product of Form A.

[1031] Fmoc-GPSSG-OH form A Preparation 1: Fmoc-GPSSG-OH Form A was prepared in 1-propanol (1-PrOH). 16 mL of 1-PrOH was added to 4.06 grams of amorphous Fmoc-GPSSG-OH solid, and the sample was stirred at ambient conditions, resulting in a reddish-orange solution. The solution was seeded with 2.8 mg of Fmoc-GPSSG-OH Form A and then left at ambient conditions with continued stirring for 2 days. A bright orange suspension was obtained, and the solid was isolated by vacuum filtration using a 10 μm disposable filter, rinsed twice on the filter with 0.5 mL of fresh 1-PrOH, then collected and dried under vacuum at 30 °C for approximately 3-4 hours. The resulting white solid (3.2 grams) was consistent with Fmoc-GPSSG-OH Form A.

[1032] Preparation 2. Fmoc-GPSSG-OH Form A was prepared in 1-propanol (1-PrOH) and heptane. Approximately 50 mg of amorphous Fmoc-GPSSG-OH was dissolved in 0.6 mL of 1-PrOH to form a clear, yellow solution. A 30 μL aliquot of the solution was added to 0.6 mL of heptane to obtain a clear, pale yellow solution. With stirring, an additional 0.6 mL of heptane was added to the solution, and the sample was capped and stirred at ambient conditions for 2 days. A suspension was obtained, and the solid was consistent with Fmoc-GPSSG-OH Form A.

[1033] Preparation 3: Fmoc-GPSSG-OH Form A was prepared in 1-propanol (1-PrOH). Approximately 10 mg of amorphous Fmoc-GPSSG-OH was dissolved in 0.1 mL of 1-PrOH to form a clear, yellow solution. The solution was stored in a capped vial at ambient conditions for 1 day and then transferred to a freezer for 3 days. The solid observed in the solution was consistent with Fmoc-GPSSG-OH Form A.

[1034] Fmoc-GPSSG-OH form B The solid Fmoc-GPSSG-OH Form A was placed in a clean TGA pan and heated to 120° C. and held at that temperature for 3 minutes in the TGA oven. The resulting white solid was consistent with Fmoc-GPSSG-OH Form B.

[1035] XRPD of Fmoc-GPSSG-OH Form A The XRPD pattern of crystalline Fmoc-GPSSG-OH Form A was obtained on a Bruker D8 Endeavor X-ray powder diffractometer equipped with a CuKα (1.5418 Å) source and a Linxeye detector, operating at 40 kV and 40 mA. The sample was scanned from 4 to 42° 2θ, with a step size of 0.009° 2θ and a scan rate of 0.5 s / step, using a 0.3° primary slit aperture and a 3.9° particle size distribution (PSD) aperture. The powder was packed into a quartz sample holder, and a smooth surface was obtained using a glass slide. Diffraction patterns were collected at ambient temperature and relative humidity. Crystalline peak positions were determined using MDI-Jade v7.9.9.

[1036] A prepared sample of Fmoc-GPSSG-OH Form A is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 38 below, specifically, a peak at 6.1° 2-theta in combination with one or more peaks selected from 8.5, 11.7, 12.3, and 16.9° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of Y-Aib-QG Form A is shown in Figure 15A.

[1037] [Table 41]

[1038] Fmoc-GPSSG-OH Form B XRPB The XRPD pattern of crystalline Fmoc-GPSSG-OH form B was obtained using the same procedure as for Fmoc-GPSSG-OH form A, except that the Fmoc-GPSSG-OH form B sample was scanned between 4 and 30 2θ° at a scan rate of 0.25 s / step.

[1039] A prepared sample of Fmoc-GPSSG-OH Form B is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 39 below, specifically, a peak at 7.2° 2-theta in combination with one or more peaks selected from the group consisting of 5.3, 8.1, 14.4, and 16.2° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of Fmoc-GPSSG-OH Form B is shown in Figure 15B.

[1040] [Table 42]

[1041] Example 13: GGG side chain

[1042] [ka]

[1043] Method of preparation, crystallization and use 2-(2-(2-aminoethoxy)ethoxy)acetic acid (1.16 equivalents) and 5 volumes of acetonitrile were placed in a jacketed reactor. N-methyl-N-trimethylsilylacetamide (2.56 equivalents) was slowly added to it. It was stirred at 20-25 °C for about 2-3 hours. The reaction mixture was clear, and 1 equivalent of (O1-tert-butyl O5-(2,5-dioxopyrrolidin-1-yl)(2S)-2-[(20-tert-butoxy-20-oxo-icosanoyl)amino]pentanedioate) was added to the reaction mixture. The reaction mixture was stirred at about 20-25 °C for 4-5 hours. Approximately 9 volumes of 2-methyltetrahydrofuran (2-Me-THF) was added to the reaction mixture, followed by extraction with 3 × 4 volumes of 2% KHSO4 and 1% NaCl aqueous solution. The organic solution was then washed with 4 × 4 volumes of 2% aqueous NaCl. The organic solution was concentrated in vacuo to reduce the acetonitrile (<0.1%) and water (KF <0.5%). 1 volume of DMF was added to create a DMF solution of (2-[2-[2-[[(4S)-5-tert-butoxy-4-[(20-tert-butoxy-20-oxo-icosanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetic acid. The organic mixture was concentrated in vacuo to reduce the 2-Me-THF to less than 5%. 2.4 equivalents of DIEA (2-[2-[2-[[(4S)-5-tert-butoxy-4-[(20-tert-butoxy-20-oxo-icosanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetic acid. A DMF feed solution of rt-butoxy-4-[(20-tert-butoxy-20-oxo-icosanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetic acid; a DMF solution of TNTU; and a DMF solution of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-lysine hydrochloride were prepared. Feed tanks of 2-MeTHF, and aqueous 3% KHSO and 5% NaCl were also prepared.The flow rates of the (2-[2-[2-[[(4S)-5-tert-butoxy-4-[(20-tert-butoxy-20-oxo-icosanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetic acid (containing 2.4 equivalents of DIEA) solution and the TNTU solution were adjusted to 0.97 relative to the (2-[2-[2-[[(4S)-5-tert-butoxy-4-[(20-tert-butoxy-20-oxo-icosanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetic acid. The reaction mixture was warmed to about 30°C in the reactor coil, and then introduced into a stream of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-lysine hydrochloride in DMF to react with 1.15 equivalents of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-lysine hydrochloride in the reactor coil at about 30°C to give the GGG side chain product. The reaction solution was then diluted with 30 volumes of 2-MeTHF and diluted with 20 volumes of 3% KHSO4 and 5% aqueous NaCl. The organic solution was then cooled to 10-20°C and the aqueous phase was separated. The organic solution was concentrated to 11-13 volumes, 2.3-2.7 volumes of DMF was added, and then extracted with 5 volumes of 3% KHSO4 and 5% NaCl aqueous solution. The organic phase was diluted with 2.3-2.7 volumes of DMF and extracted with 5 volumes of 3% KHSO4 and 5% NaCl aqueous solution. Next, 2.5-3.2 volumes of 2-MeTHF and 2.3-2.7 volumes of DMF were added, and the organic matter was extracted with 5 volumes of 3% KHSO4 and 5% NaCl aqueous solution. The organic solution was diluted with DMF and extracted with 3% KHSO4 and 5% NaCl, followed by another cycle of dilution with 2-MeTHF and DMF, followed by extraction with 3% KHSO4 and 5% NaCl. 2.5-3.2 volumes of 2-MeTHF were then added, and the organic solution was extracted four times with 5% aqueous NaCl. The 2-MeTHF solution was then concentrated, and the 2-MeTHF was exchanged for acetonitrile by repeated addition of acetonitrile and concentration of the solution under vacuum until the 2-MeTHF concentration was reduced to below 10%.

[1044] Crystallization After solvent exchange by distillation, ACN was added to 30 volumes. The solution was heated to 55°C to completely dissolve all side chain material. The solution was cooled to 5°C over 4 hours and then held for 4 hours for primary nucleation. Primary nucleation occurred reliably during the 5°C hold, resulting in a mixture of amorphous and crystalline material. As this was undesirable, the next step was designed to convert to crystalline material.

[1045] The solution was warmed to 30° C. and held for 8 hours to dissolve the amorphous material. Next, 6 volumes of MEK was added and the solution was held for an additional 3 hours to dissolve additional amorphous material, then cooled to 20° C. Four thermal cycles were used to convert from amorphous to crystalline. In each cycle, the mixture was heated to 30° C. and held for 3 hours, then cooled to 20° C. and held for 3 hours.

[1046] After thermal cycling, the mixture was cooled to 5°C over 1 hour and held for 1 hour. A sample of the supernatant was collected and tested by HPLC for side chain potency. A side chain concentration of 5 mg / mL or less indicated that the solid form was a low-solubility crystalline form. The slurry was then filtered, rinsed three times with 10 volumes of ACN at 5°C, and dried at 30°C.

[1047] GGG side chain crystalline form Three crystalline solid forms of the GGG side chain have been identified, including GGG SC Form A, Form B, and Form C.

[1048] Form A is a crystalline anhydrous / non-solvated form that is observed from a variety of solvent conditions at ambient temperature.

[1049] Form B is a crystalline material and has been observed in water.

[1050] Form C is a crystalline anhydrous / non-solvated form and was observed at elevated temperatures.

[1051] GGG SC form A 241.2 mg of solid GGG side chain was added to 2 mL of 1:3 vol / vol MEK / ACN, and the mixture was heated at approximately 75 °C for 10 min to give a clear solution, which remained clear after slowly cooling to ambient temperature.

[1052] Approximately half of the above solution was seeded with disordered Form A and then stirred at ambient conditions, producing first gel particles and then a thick white paste with no mobile liquid. After adding an additional 0.5 mL of 1:3 vol / vol MEK / ACN, the sample was stirred at ambient conditions for 2 days to yield a thick white slurry. The white solid isolated from the sample by vacuum filtration was consistent with GGG side-chain Form A.

[1053] GGG side chain Form A was also obtained by stirring the above solution at ambient conditions without seeding. The sample also underwent the formation of gel particles and a thick white paste, which required the addition of an additional volume of 1:3 vol / vol MEK / ACN to slurry.

[1054] GGG side chain form B Approximately 20 mg of the disordered GGG side chain solid was slurried in water at 60 °C to yield birefringent white particles. The sample was isolated and dried on a Whatman filter under a stream of nitrogen. The resulting white solid was consistent with GGG side chain Form B.

[1055] GGG side chain form C GGG side chain form A was heated to 93°C and the resulting solid was consistent with GGG side chain form C.

[1056] XRPD patterns of the crystalline GGG side chain (Form A or Form B) were obtained on a Bruker D8 Endeavor X-ray powder diffractometer equipped with a CuKα (1.5418 Å) source and a Linxeye detector, operating at 40 kV and 40 mA. Samples were scanned from 4 to 42° 2θ using a 0.009° 2θ step size and a 0.5 s / step scan rate, a 0.3° primary slit aperture, and a 3.9° particle size distribution (PSD) aperture. In some cases, scan ranges of 4 and 30° 2θ were used, with a 0.009° 2θ step size and a 0.25 s / step scan rate. Samples were packed into quartz or silicon sample holders, and smooth surfaces were obtained using glass slides. Crystalline form diffraction patterns were collected at ambient temperature and relative humidity. Crystalline peak positions were determined using MDI-Jade v7.9.9. It is well known in the field of crystallography that for any crystalline form, the relative intensities of diffraction peaks may vary due to preferred orientation resulting from factors such as crystalline form and crystal habit. When the effect of preferred orientation is present, peak intensities change, but the characteristic peak positions of a polymorph remain unchanged. See, for example, The United States Pharmacopeia #23, National Formulary #18, pages 1843-1844, 1995. It is also well known in the field of crystallography that for any crystalline form, angular peak positions may vary slightly. For example, peak positions may shift due to variations in the temperature at which the sample is analyzed, sample displacement, or the presence or absence of an internal standard. In this case, a peak position variation of ±0.2 2θ° is estimated to account for these potential variations without precluding unambiguous identification of the indicated crystalline form. Confirmation of a crystalline form can be based on any unique combination of characteristic peaks.

[1057] The XRPD pattern of crystalline GGG side chain Form C was obtained using a PANalytical X'Pert PRO MPD diffractometer with an incident beam of Cu Kα radiation generated using a long, fine-focus source and a nickel filter. The diffractometer was configured using a symmetric Bragg-Brentano geometry. Data were collected and analyzed using Data Collector software v.2.2b. Prior to analysis, a silicon specimen (NIST SRM 640e) was analyzed to verify that the position of the observed Si 111 peak matched the NIST-certified position. Sample pieces were loaded into nickel-coated copper wells. Anti-scatter slits were used to minimize background generated by air scattering. Soller slits for the incident and diffracted beams were used to minimize off-axis divergence. Diffraction patterns were collected using a scanning position-sensitive detector (X'Celerator) positioned 240 mm from the sample. Data acquisition was from 3.5° to 30° 2θ with a step size of 0.017° 2θ. Crystalline peak positions were determined using MDI-Jade v7.9.9. It is well known in the field of crystallography that for any crystalline form, the relative intensities of diffraction peaks may vary due to preferred orientation resulting from factors such as crystalline form and crystal habit. When the effect of preferred orientation is present, peak intensities change, but the characteristic peak positions of polymorphs remain unchanged. See, for example, The United States Pharmacopeia #23, National Formulary #18, pages 1843-1844, 1995. It is also well known in the field of crystallography that for any crystalline form, angular peak positions may vary slightly. For example, peak positions may shift due to variations in the temperature at which the sample is analyzed, sample displacement, or the presence or absence of an internal standard. In this case, a peak position variation of ±0.2 2θ° is estimated to account for these potential variations without precluding unambiguous identification of the indicated crystalline form. Confirmation of a crystalline form can be based on any unique combination of characteristic peaks.

[1058] XRPD of GGG side chain, Form A A prepared sample of GGG side chain, Form A, is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 40 below, specifically, a peak at 11.4° 2-theta in combination with one or more peaks selected from the group consisting of 6.0, 8.9, 12.7, 13.6, 14.6, 17.0, and 18.8° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of GGG side chain Form A is shown in Figure 16A.

[1059] [Table 43]

[1060] XRPD of GGG side chain form B A prepared sample of GGG side chain, Form B, is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 41 below, specifically, a peak at 10.6° 2-theta in combination with one or more peaks selected from the group consisting of 7.1, 12.1, 13.6, 14.2, 15.2, 16.0, and 16.8° 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of GGG side chain Form B is shown in Figure 16B.

[1061] [Table 44]

[1062] XRPD of GGG side chain form C A prepared sample of GGG side chain, Form C, is characterized by an XRPD pattern using CuKα radiation as having diffraction peaks (2-theta values) as set forth in Table 42 below, specifically peaks at 10.1 and 15.5 degrees 2-theta, in combination with one or more peaks selected from the group consisting of 6.1, 8.7, 11.4, 16.6, and 19.2 degrees 2-theta, with a diffraction angle tolerance of 0.2 degrees. A representative XRPD pattern of GGG side chain Form C is shown in Figure 16C.

[1063] [Table 45]

[1064] Example 14: H-Ala-Pro-Pro-Pro-Ser(tBu)-NH

[1065] [ka]

[1066] Crystallization studies for H-Ala-Pro-Pro-Pro-Ser(tBu)-NH2 (HAPPPS-NH2) were carried out on the free form (free base) and via salt / co-crystalline formation.

[1067] Crystallization of the free form was attempted in a variety of solvents and solvent mixtures using different techniques, such as slurry, cooling, and holding at ambient and subambient temperatures. Crystalline HAPPPS-NH2 was not identified under the conditions investigated.

[1068] Crystallization by salt formation was also performed. A total of 96 conditions, including 12 counterions, each in eight different solvents, were tested at a ratio of approximately 1:1.2 mol / mol (peptide / acid). The counterions and solvents used in the screening are listed in Table 43 below. Crystalline mono-HCl salts were observed in ACN (wet conditions only) and acetone.

[1069] [Table 46]

[1070] Crystallization by co-crystal formation was carried out in CPME. Amorphous HAPPPS-NH2 solid was stirred overnight at ambient conditions in a CPME solution saturated with the co-former. 96 co-formers, listed in Table 44 below, were investigated. No crystalline HAPPPS-NH2 co-crystals were identified under the conditions tested.

[1071] [Table 47-1]

[1072] [Table 47-2]

[1073] Two crystalline solid forms of the HCl salt of HAPPPS-NH2 tetramer have been identified, including HAPPPS-NH2HCl Form A and Form B.

[1074] HAPPPS-NH2HCl Form A is produced by salt reaction in acetonitrile, which becomes amorphous upon isolation / drying. HAPPPS-NH2HCl Form B is produced by salt reaction in acetone. Both are mono-HCl salts based on ion chromatography analysis.

[1075] HAPPPS-NH2HCl Form A Lot 1 One mL of acetonitrile (ACN) was added to 79.4 mg of the amorphous solid HAPPPS-NH2, producing a clear, colorless solution at ambient conditions. While stirring, 183 μL of 1 N HCl in EtOAc (approximately 1:1.2 mol / mol peptide:HCl) was added to the peptide solution. The sample was stirred at ambient conditions, and the entire sample quickly filled with a white solid, which appeared as thin, hair-like, birefringent needles under a polarized light microscope. An additional 3 mL of ACN was added to the sample, resulting in a white slurry. The wet solid from the slurry was consistent with HAPPPS-HCl Form A, which lost its crystallinity upon isolation and drying.

[1076] Lot 2 3.1 grams of amorphous HAPPPS-NH2TFA salt was dissolved in 50 mL of ACN to give a clear, yellow solution. With stirring, 8 mL of 1N HCl in EtOAc was added to the peptide solution. The sample was stirred at ambient conditions, and a white precipitate appeared immediately after stirring until the entire sample became a thick, off-white slurry. An additional 150 mL of ACN was added to the sample and stirred at ambient conditions for 2 days. The wet solid from the slurry was consistent with HAPPPS-HCl Form A.

[1077] The solid from the slurry was isolated by vacuum filtration using a stream of N2 on top of the filter, rinsed on the filter with fresh ACN, then collected and dried under vacuum at 30° C. overnight. The resulting white solid was amorphous.

[1078] HAPPPS-NH2HCl Form B One mL of acetone was added to 89.3 mg of the amorphous solid HAPPPS-NH2, producing a clear, colorless solution at ambient conditions. With stirring, 205 μL of 1 N HCl in EtOAc (approximately 1:1.2 mol / mol peptide:HCl) was added to the peptide solution, resulting in a thick, white slurry. An additional 2 mL of acetone was added, and the sample was stirred at ambient conditions for 2 days. The solid from the slurry was isolated by vacuum filtration with a stream of N2 on top of the filter, and the resulting white solid (55 mg) was consistent with H HAPPPS-HCl Form B.

[1079] XRPD of HAPPPS-NH2HCl Form A The XRPD pattern of crystalline HAPPPS-NH2HCl was obtained on a PANalytical Empyrean diffractometer equipped with a CuKα (1.5418 Å) source and a PIXcel3D 1x1 detector, operating at 45 kV and 40 mA. The sample was scanned between 2 and 40 2θ° with a step size of 0.0065652 2θ° for a total of 5788 steps over 3725 seconds. The sample was sandwiched between Etnom films in the sample holder to prevent solvent evaporation and analyzed in transmission geometry. Diffraction patterns were collected at ambient temperature.

[1080] A prepared sample of HAPPPS-NHHCl Form A was characterized by its XRPD pattern using CuKα radiation as described above. The sample is characterized as having the diffraction peaks (2-theta values) set forth in Table 45 and shown in Figure 17A, specifically, a peak at 5.0° 2-theta in combination with one or more peaks selected from 8.3, 9.7, and 11.2° 2-theta, with a diffraction angle tolerance of 0.2 degrees.

[1081] [Table 48]

[1082] XRPD of dried solid from HAPPPS-NH2HCl Form A HAPPPS-NHHCl Form A loses crystallinity upon isolation / drying. Dried samples exhibited XRPD patterns with broad halos with or without broad peaks at low angles, as shown in Figure 17B.

[1083] XRPD of HAPPPS-NH2HCl Form B A prepared sample of HAPPPS-NHHCl Form B was characterized by its XRPD pattern using CuKα radiation as described above. The sample is characterized as having the diffraction peaks (2-theta values) set forth in Table 46 and shown in Figure 17C, specifically, a peak at 7.2° 2-theta in combination with one or more peaks selected from 5.3, 8.1, 14.4, and 16.2° 2-theta, with a diffraction angle tolerance of 0.2 degrees.

[1084] [Table 49]

[1085] Example 15: Fmoc-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-NH

[1086] [ka]

[1087] Crystallization studies of Fmoc-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-NH2 (Fmoc-SSGAPPPS-NH2) were performed via salt / co-crystal formation. One solid form, Fmoc-SSGAPPPS-NH2 Form A, was identified. Form A represents a family of isostructural solvates and can be generated from multiple solvent conditions, including acetone, acetonitrile (ACN), methyl acetate (MeOAc), and methyl ethyl ketone (MEK).

[1088] Lot 1 0.4 mL of acetone was added to approximately 60 mg of amorphous Fmoc-SSGAPPPS-NH2 solid, resulting in a clear, yellow solution. The solution was stirred at ambient conditions for 2 days, resulting in a white slurry. The solid isolated from the slurry was consistent with Fmoc-SSGAPPPS-NH2 Form A. Following a similar procedure, Fmoc-SSGAPPPS-NH2 Form A was also prepared from other organic solvents, including acetonitrile (ACN), methyl acetate (MeOAc), and methyl ethyl ketone (MEK).

[1089] Lot 2 3.09 grams of amorphous Fmoc-SSGAPPPS-NH2 solid was dissolved in 15 mL of MeOAc. The solution was seeded with Fmoc-SSGAPPPS-NH2 Form A solid and stirred at ambient conditions for 2 days. An additional 12 mL of MeOAc was added during the process to facilitate stirring. The white solid was isolated from the slurry by vacuum filtration using a Whatman paper filter, rinsed four times with 3 mL of fresh MeOAc on the filter, then collected and dried under vacuum at 30 °C for approximately 3 hours. The resulting white solid (2.4 grams) was consistent with Fmoc-SSGAPPPS-NH2 Form A.

[1090] XRPD of Fmoc-SSGAPPPS-NH2 Form A The XRPD pattern of crystalline Fmoc-SSGAPPPS-NH2 Form A was obtained on a Bruker D8 Endeavor X-ray powder diffractometer equipped with a CuKα (1.5418 Å) source and a Linxeye detector, operating at 40 kV and 40 mA. The sample was scanned from 4 to 42° 2θ, with a step size of 0.009° 2θ and a scan rate of 0.5 seconds per step, using a 0.3° primary slit aperture and a 3.9° PSD aperture. The powder was packed into a quartz sample holder, and a glass slide was used to obtain a smooth surface. The crystalline form diffraction pattern was collected at ambient temperature and relative humidity. Crystalline peak positions were determined using MDI-Jade v7.9.9.

[1091] A prepared sample of Fmoc-SSGAPPPS-NH2 Form A was characterized by its XRPD pattern using CuKα radiation as described above. The sample is characterized as having the diffraction peaks (2-theta values) set forth in Table 47 and shown in Figure 18, specifically, as having a peak at 7.5° 2-theta in combination with one or more peaks selected from 6.1, 8.7, 10.6, 15.0, 16.1, and 18.6° 2-theta, with a diffraction angle tolerance of 0.2 degrees.

[1092] [Table 50]

[1093] Example 16: Liquid Phase Peptide Synthesis (LPPS) of Tirzepatide (TZP) Fragment 1 Tirzepatide fragment 1 (TZP fragment 1) is a peptide consisting of 10 amino acids (GPSSGAPPPS). A solution-phase peptide synthesis (LPPS) strategy for preparing TZP fragment 1 was tested using the TFA salt of the compound (TFA NH-GPSSGAPPS-CONH). LPPS synthesis of TZP fragment 1 was achieved by coupling Fmoc-GP-OH and NH-GPSSGAPPS-CONH, followed by deprotection of the Fmoc group and precipitation of the product with a known amount of trifluoroacetic acid.

[1094] material Fmoc-GP-OH, EDC.HCl, oxyma, diisopropyl ethyl ether (DIPEA), acetonitrile (ACN), isopropyl acetate (IPAc), trifluoroacetic acid (TFA), ethyl acetate (EtOAc), sodium bicarbonate (NaHCO), diethylamine (DEA), methyl tert-butyl ether (MTBE), heptane, and sodium sulfate were purchased and used without any purification. NH-GPSSGAPPS-CONH was synthesized by solid-phase peptide synthesis (SPPS).

[1095] Synthesis of Fmoc-GPSSGAPPS-CONH2 - Fmoc Coupling The synthesis of Fmoc-GPSSGAPPS-CONH2 (protected TZP fragment 1) was achieved by coupling Fmoc-GP-OH to NH2-GPSSGAPPS-CONH2 in acetonitrile / isopropyl acetate (ACN / IPAc).

[1096] [ka]

[1097] To a round-bottom flask equipped with a stir bar, 20 mL (20 V) of ACN / IPAc (1:3), Fmoc-GP-OH (1.00 equiv., 1.16 mmol, 456 mg), and oxyma (1.12 equiv., 1.29 mmol, 184 mg) were added with stirring at room temperature. EDC.HCl (1.26 equiv., 1.46 mmol, 279 mg) and DIPEA (1.0 equiv., 1.16 mmol, 199 μL) were added and stirred at room temperature for 10 min for preactivation (the reaction turned yellow). After sonicating the reaction flask for 1 min, NH2-GPSSGAPPS-CONH2 (1.00 equiv., 1.16 mmol, 1.00 g) was added in one portion and stirred for 1 h. The pH of the reaction was maintained between 3.5 and 5.5. Upon completion, the crude mixture was transferred to a separatory funnel and the flask was rinsed with 2 ml of ethyl acetate (EtOAc) and transferred to the separatory funnel. The mixture was washed twice with 7 ml (7 V) of 1 M aqueous HCl and the aqueous layer was separated into Erlenmeyer flask 1. This was followed by two base washes with 7 ml (7 V) each of saturated aqueous NaHCO3 and the aqueous layer was drained into Erlenmeyer flask 2. It was then washed twice with 7 ml of DI water and the aqueous layer was drained into Erlenmeyer flask 2. The organic layer was transferred to Erlenmeyer flask 3.

[1098] 20 ml (20 V) of fresh ethyl acetate was added to the separatory funnel to back-extract the aqueous acid and base layers. The organic layers were combined, dried over Na2SO4, and filtered into a flask. The solvent was removed by rotary evaporation to give Fmoc-GPSSGAPPS-CONH2 as a solid, 1.032 g (70% yield).

[1099] Synthesis of F1-NH2.TFA - Fmoc deprotection and subsequent TFA salification

[1100] [ka]

[1101] In a flask containing a stir bar, Fmoc-GPSSGAPPS-CONH2 (1.0 equiv., 0.79 mmol, 0.98 g) was dissolved in ACN (3 V, 2.92 ml) at room temperature. Diethylamine (10 equiv., 0.812 ml) was added and stirred at room temperature for 1 hour. The solvent was reduced to 2 ml (approximately 2 V) by rotary evaporation, and 15 ml (approximately 15 V) of chilled 1:1 MTBE:heptane was added slowly with stirring to precipitate the product, which was then filtered. The flask was rinsed with additional chilled 1:1 MTBE:MTBE. Heptane was added and the mixture was filtered.

[1102] The combined solids were dissolved in 2 ml of ACN, transferred to a vial, and dried completely by rotary evaporation. 2 ml of ACN and a stir bar were added to the material in the vial, and trifluoroacetic acid (15 equiv., 11.8 mmol, 0.904 ml) was added in one portion at room temperature. The reaction was stirred for 50 minutes, the stir bar was removed, and 15 ml of chilled 1:1 MTBE:MTBE was added. Heptane was added slowly with stirring to give a suspension that was filtered to give the solid product. The combined solids were dried by rotary evaporation to give F1-NH2.TFA as an off-white solid, 0.87 g (77% yield).

[1103] Screening Coupling Agents Different coupling agents were screened for the synthesis of Fmoc-GPSSGAPPS-CONH2 by coupling of Fmoc-GP-OH to NH2-GPSSGAPPS-CONH2. A summary of the coupling agents screened for the reaction is shown in Table 48.

[1104] [Table 51]

[1105] In the experiment, a total of 24 coupling agents were screened for the reaction to determine which would be best for complete conversion of starting materials to product in the shortest time, reduced impurity formation, ease of impurity removal by extraction, cost-effectiveness, and minimal health hazard.

[1106] [ka]

[1107] Coupling agent screening was performed using a "Dreadnought" instrument set. Fmoc-GP-OH and each coupling agent were dispensed into the appropriate wells using a Quantos system. Stock solutions of 2,4,6-collidine in ACN, DEPBT / 2,4,6-collidine in ACN, and T3P / 2,4,6-collidine in ACN were prepared and dispensed into the appropriate wells in the glovebox. The plate was sealed, removed from the glovebox, and agitated on a polyblock at 800 rpm for 30 minutes before being returned to the glovebox, where a stock solution of NH2-GPSSGAPPS-CONH2 was prepared and 1.1 equivalents were dispensed into each vial.

[1108] The plate was removed from the glove box and stirred at room temperature for 4 hours. LCMS analytical samples were taken from each vial at 1 hour and 4 hour intervals, respectively, diluted, and analyzed by LCMS for conversion at the 1 hour and 4 hour marks. The results are shown in Figure 19.

[1109] Of the 24 coupling agents screened for the reaction, 15 showed 100% product formation (100% conversion of limited starting material to product) within a 1-hour interval, while 2 coupling agents showed 100% product formation only at a 4-hour interval. Seven coupling agents did not achieve complete conversion of limited starting material to product, even at a 4-hour reaction time.

[1110] Example 17: Solvent Screening Test Peptide samples were screened for crystal stability in several organic solvents. Four peptide tetramers were used: (i) Boc-Y(tBu)-Aib-E(Me)-G-COOH; (ii) Boc-Y(tBu)-Aib-E(All)-G-COOH; (iii) Boc-Y(tBu)-Aib-E(cHx)-G-COOH; and (iv) Boc-Y(tBu)-Aib-E(Bzl)-G-COOH. The following organic solvents were used: acetone, ACN, MeOH, EtOH, IPA, BuOH, EtOAc, MTBE, THF, MEK, toluene, and water.

[1111] A 30 mg sample of each peptide tetramer was dispensed into sample vials (one vial of each peptide for testing in e...

Claims

1. A compound of formula (I) or a salt, solvate, or hydrate thereof: 【Chemical 1】 [In the formula, R １ is H or a protecting group, and R ２ is H or a protecting group, and R ３ is H or a protecting group, and R ４ is H or a protecting group, and R １ , R ２ , R ３ , and R ４ at least one of which is a protecting group.

2. a) R1 is H, Fmoc, or Boc; b) R2 is H or t-butyl; c) R4 is H or benzyl, or d) The compound of claim 1, which is any combination of (a) to (c).

3. R ３ The compound of claim 1 or 2, wherein is H, t-butyl, benzyl, carboxybenzyl, methyl, allyl, cyclohexyl, or trityl.

4. (a) R １ , R ２ , R ３ , and R ４ are H, or (b) R １ , R ２ , R ３ , and R ４ The compound of any one of claims 1 to 3, wherein at least two of (a) and (b) are protecting groups, or (c) both (a) and (b).

5. The compound is 【Chemistry 2】 2. The compound of claim 1, wherein the compound is selected from the formula:

6. (i) the compound is a solvate, and optionally the solvate is formed from methyl t-butyl ether (MTBE) or a mixture containing MTBE, or (ii) the compound is crystalline, and optionally a) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.2°±0.2 degrees in combination with one or more peaks selected from 8.4°, 8.8°, 10.4°, 15.5°, 17.1°, and 17.7°; or b) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 6.1°±0.2 degrees in combination with one or more peaks selected from 10.3°, 14.9°, 16.8°, 18.1°, and 18.2°; or c) The compound according to any one of claims 1 to 5, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 6.2° ± 0.2 degrees in combination with one or more peaks selected from 8.9°, 12.3°, 14.9°, 15.4°, and 21.8°.

7. 10. A method for synthesizing a compound of claim 1, comprising: Formula (Y 保護 ) and 【Chemistry 3】 [In the formula, R １ and R ２ is a protecting group], 保護 ) compounds 【Chemistry 4】 [In the formula, R ５ is a protecting group] and a compound of formula (Aib 保護 ) to react with a compound of formula (Y 保護 -Aib 保護 and forming a compound of formula (II) 【Chemistry 5】 -R ５ and removing the protecting group of formula (Y 保護 -Aib), and 【Chemistry 6】 - the formula (Y 保護 -Aib) and a compound of formula (E 保護 ) compounds 【Chemistry 7】 [In the formula, R ３ is a protecting group] to form a compound of the formula (Y 保護 -Aib-E 保護 and forming a compound of formula (II) 【Chemistry 8】 - the formula (Y 保護 -Aib-E 保護 ) and a compound of formula (G 保護 ) compounds 【Chemistry 9】 [In the formula, R ４ is a protecting group] to form a compound of the formula (Y 保護 -Aib-E 保護 -G 保護 and forming a compound of formula (II) 【Chemistry 10】 and, optionally, -Protecting group R １ , R ２ , R ３ , and R ４ and removing one or more of: A method comprising:

8. 10. A method of synthesizing a polypeptide, wherein (i) the polypeptide is SEQ ID NO: 1, the method comprising conjugating a compound of any one of claims 1 to 5 via the C-terminus of the compound to the N-terminus of a polypeptide of SEQ ID NO: 2, or (ii) the polypeptide is SEQ ID NO: 26, the method comprising conjugating a compound of any one of claims 1 to 5 via the C-terminus of the compound to the N-terminus of a polypeptide of SEQ ID NO:

27.

9. A compound of formula (II) or a salt, solvate, or hydrate thereof: 【Chemistry 11】 [In the formula, R ６ is H or a protecting group, and R ７ is H or a protecting group, and R ８ is H or a protecting group, and R ９ is H or a protecting group, and R １０ is H or a protecting group, optionally each protecting group independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

10. a) R ６ is H, Fmoc, or Boc; b) R ７ is H or t-butyl, c) In the formula, R ８ is H or t-butyl, d) R ９ is H or t-butyl, e) R １０ is H or benzyl, or f) The compound according to claim 9, which is any combination of (a) to (e).

11. (a) R ６ , R ７ , R ８ , R ９ , and R １０ is a protecting group, or (b) R ６ , R ７ , R ８ , R ９ , and R １０ 11. The compound of claim 9 or 10, wherein is H.

12. The compound has the formula: 【Chemistry 12】 10. The compound of claim 9, wherein

13. (i) the compound is a solvate, optionally the solvate is produced from a solvent comprising heptane, or (ii) the compound is crystalline, optionally a) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 7.7 to 7.9° ± 0.2 degrees in combination with one or more peaks selected from 5.8°, 10.0°, 10.8-10.9°, 11.3-11.4°, 12.0-12.1°, 12.8°, 14.2-14.4°, and 16.8-17.0°; or b) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 7.3° ± 0.2 degrees in combination with one or more peaks selected from 5.1°, 5.7°, 7.6°, 9.5°, and 12.4°; or c) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 7.8° ± 0.2 degrees in combination with one or more peaks selected from 8.5°, 11.5°, 12.0°, 12.8°, 14.3°, 15.5°, 20.2°, and 23.3°; or d) The compound according to any one of claims 9 to 12, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 8.2 to 8.3° ± 0.2 degrees in combination with one or more peaks selected from 5.9°, 7.7°, 9.2°, 10.2°, 11.3°, 13.8 to 13.9°, 15.5 to 15.7°, 17.1°, and 18.5°.

14. 13. A method of synthesizing a polypeptide of SEQ ID NO: 1, comprising: (i) conjugating a compound of any one of claims 9 to 12 via the N-terminus of the compound to the C-terminus of a polypeptide of SEQ ID NO: 3 to form a polypeptide of SEQ ID NO: 4; and (ii) conjugating the polypeptide of SEQ ID NO: 4 via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO:

5.

15. A compound of formula (III) or a salt, solvate, or hydrate thereof, 【Chemistry 13】 In the formula, R １１ is H or a protecting group, and R １２ is H or a protecting group, optionally each protecting group independently selected from Boc, Fmoc, tert-butyl, and trityl groups, or a salt, solvate, or hydrate thereof.

16. a) R １１ is H or t-butyl, b) R １２ is H or t-butyl, or c) The compound of claim 15, which is both (a) and (b).

17. (a) R １１ and R １２ is a protecting group, or (b) R １１ and R １２ 17. The compound of claim 15 or 16, wherein is H.

18. The compound has the formula: 【Chemistry 14】 16. The compound of claim 15,

19. (i) the compound is a solvate, or (ii) the compound is crystalline, and optionally a) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 8.0° ± 0.2 degrees in combination with one or more peaks selected from 7.0°, 10.3°, 14.1°, 15.2°, 16.7°, 18.0°, 19.0°, 19.7°, 20.8°, and 21.9°; or b) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 9.0° ± 0.2 degrees in combination with one or more peaks selected from 5.7°, 9.9°, 16.2°, 17.1°, 17.9°, 18.1°, 18.4°, 18.8°, 19.9°, 20.1°, and 22.5°; or c) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 10.5° ± 0.2 degrees in combination with one or more peaks selected from 5.6°, 11.9°, 13.3°, 15.4°, 15.6°, 18.1°, 19.9°, and 21.1°; or d) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 10.7°±0.2 degrees in combination with one or more peaks selected from 4.9°, 14.8°, 20.3°, and 21.5°; or e) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 10.5° ± 0.2 degrees in combination with one or more peaks selected from 5.9°, 10.5°, 10.9°, 12.1°, 13.1°, 15.9°, 17.5°, 20.9°, 21.1°, and 21.9°; or f) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 7.8°±0.2 degrees in combination with one or more peaks selected from 11.3°, 11.5°, 15.4°, 15.6°, and 21.5°; or g) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 10.0° ± 0.2 degrees in combination with one or more peaks selected from 8.1°, 12.5°, 13.5°, 14.7°, 17.8°, 18.8°, 20.0°, and 22.4°; or h) The compound according to any one of claims 15 to 18, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 21.1° ± 0.2 degrees in combination with one or more peaks selected from 5.6°, 10.5°, 10.8°, 11.9°, 15.4°, and 23.8°.

20. 19. A method of synthesizing a polypeptide of SEQ ID NO: 1, comprising: (i) conjugating a compound of any one of claims 15 to 18 via the N-terminus of the compound to the C-terminus of a polypeptide of SEQ ID NO: 6 to form a polypeptide of SEQ ID NO: 7; and (ii) conjugating the polypeptide of SEQ ID NO: 7 via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO:

8.

21. A compound of formula (IV) or a salt, solvate, or hydrate thereof: 【Chemistry 15】 [In the formula, R １３ is H or a protecting group, and R １３＊ is H or a protecting group, and R １４ is H or a protecting group, and R １５ is H or a protecting group, optionally each protecting group independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

22. a) R １３ is H, Fmoc, or Boc; b) R １３＊ is H, c) R １４ is H or t-butyl, d) R １５ is H or t-butyl, or e) The compound of claim 21, which is any combination of (a) to (d).

23. (a) R １３ , R １３＊ , R １４ , and R １５ is a protecting group, or (b) R １３ , R １３＊ , R １４ , and R １５ 23. The compound of claim 21 or 22, wherein is H.

24. The compound has the formula: 【Chemistry 16】 22. The compound of claim 21,

25. (i) the compound is a solvate; (ii) the compound is crystalline; and optionally, a) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.1° ± 0.2 degrees in combination with one or more peaks selected from 4.3°, 6.1°, 8.0°, 10.1°, and 18.7°; or b) The compound according to any one of claims 21 to 24, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.2° ± 0.2 degrees in combination with one or more peaks selected from 6.0°, 6.7°, 10.0°, 10.3°, 16.4°, 17.8°, 18.3°, 19.4°, and 22.4°.

26. 25. A method of synthesizing a polypeptide of SEQ ID NO: 1, comprising: (i) conjugating a compound of any one of claims 21 to 24 via the N-terminus of the compound to the C-terminus of a polypeptide of SEQ ID NO: 9 to form a polypeptide of SEQ ID NO: 7; and (ii) conjugating the polypeptide of SEQ ID NO: 7 via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO:

8.

27. The compound has the formula 【Chemistry 17】 is a compound of In the formula, R ５４ is H or a protecting group, and R ５５ is H or a protecting group, and R ５６ is H or a protecting group, optionally each protecting group independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

28. a) R ５４ is H or tert-butyl, b) R ５５ is H or tert-butyl, c) R ５６ is H or Boc, or d) The compound of claim 27, which is any combination of (a) to (c).

29. (a) R ５４ , R ５５ , and R ５６ is H, or (b) R ５４ , R ５５ , and R ５６ 29. The compound of claim 27 or 28, wherein at least one of (a) and (b) is a protecting group, or (c) both (a) and (b).

30. The compound has the formula: 【Chemistry 18】 28. The compound of claim 27,

31. (i) the compound is a solvate, optionally a solvate formed from acetone, (ii) the compound is a desolvate, or (iii) the compound is crystalline, optionally a) the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 5.8° and 18.5° ±0.2 degrees in combination with one or more peaks selected from 8.6°, 9.4°, 12.9°, 13.8°, 17.2°, and 19.4°; or b) the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta ±0.2 degrees between 7.0 and 7.1 degrees and between 7.5 and 7.7 degrees in combination with one or more peaks selected from 5.3 to 5.4 degrees, 9.7 to 9.9 degrees, and 14.7 to 14.9 degrees, or c) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 8.3° ±0.2 degrees in combination with one or more peaks selected from 6.3°, 11.4°, 14.3°, and 16.6°; or d) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.2° ±0.2 degrees in combination with one or more peaks selected from 6.8°, 8.6°, 15.8°, and 18.9°; or e) The compound according to any one of claims 27 to 30, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 6.1° ±0.2 degrees in combination with one or more peaks selected from 6.3°, 7.8°, 10.0°, and 12.4°.

32. 28. A method of synthesizing a compound of claim 27, comprising: - a compound of formula (Fmoc-G) and a compound of formula (P 保護 ) to react with a compound of the formula (Fmoc-GP 保護 ) forming a compound of formula (I) 【Chemistry 19】 (In the formula, R ５７ is a protecting group) - the protecting group R ５７ to form the compound (Fmoc-GP), 【Chemistry 20】 - a compound of the formula (Fmoc-GP) and a compound of the formula (S 保護２ ) to react with a compound of the formula (Fmoc-GP-S 保護２ ) forming a compound of formula (I) 【Chemical 21】 (In the formula, R ５４ and R ５８ is a protecting group) - the protecting group R ５８ (Fmoc-GP-S 保護１ ) forming a compound of formula (I) 【Chemical 22】 -The above formula (Fmoc-G-P-S 保護１ ) and a compound of formula (S 保護２ ) to react with a compound of the formula (Fmoc-GP-S 保護１ -S 保護２ ) forming a compound of formula (I) 【Chemical 23】 (In the formula, R ５５ and R ５９ is a protecting group) - the protecting group R ５９ (Fmoc-F-G 保護１ -S 保護１ -S 保護１ ) forming a compound of formula (I) 【Chemistry 24】 -The above formula (Fmoc-G-P-S 保護１ -S 保護１ ) and a compound of formula (G 保護 ) to react with a compound of the formula (Fmoc-GP-S 保護１ -S 保護１ -G 保護 ) forming a compound of formula (I) 【Chemistry 25】 (In the formula, R ６０ is a protecting group) -The above formula (Fmoc-G-P-S 保護１ -S 保護１ -G 保護 28. Converting a compound of formula (I) to a compound of formula (I), 【Chemical 26】 and optionally one or more protecting groups R ５４ , R ５５ , and R ５６ The method of claim 1, further comprising:

33. 33. The method of claim 32, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

34. A method for synthesizing a polypeptide, (i) wherein the polypeptide is SEQ ID NO: 12, said method comprising: (i.1) conjugating a compound according to any one of claims 27 to 30 via the C-terminus of said compound to the N-terminus of a polypeptide of SEQ ID NO: 22; and (i.2) conjugating the resulting compound via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO: 23; or (ii) wherein the polypeptide is SEQ ID NO: 24, said method comprising conjugating a compound according to any one of claims 27 to 30 via the C-terminus of said compound to the N-terminus of a polypeptide of SEQ ID NO:

22. (iii) the polypeptide is SEQ ID NO: 16, and the method comprises conjugating a compound of any one of claims 27 to 30 via the N-terminus of the compound to the C-terminus of a polypeptide of SEQ ID NO: 25; or (iv) the polypeptide is SEQ ID NO: 1, and the method comprises (iv.1) conjugating a compound of any one of claims 27 to 30 via the C-terminus of the compound to the N-terminus of a polypeptide of SEQ ID NO: 8, and (iv.2) conjugating the resulting compound via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO:

9.

35. A compound of formula (V) or a salt, solvate, or hydrate thereof: 【Chemical 27】 [In the formula, R １６ is H or a protecting group, and R １７ is H or a protecting group, and R １８ is H or a protecting group, and R １９ is H or a protecting group, and R ２０ is H or a protecting group, and R ２１ is H or a protecting group, optionally each protecting group independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

36. a) R １６ is H, Fmoc, or Boc; b) R １７ is H or t-butyl, c) R １８ is H or t-butyl, d) R １９ is H or t-butyl, e) R ２０ is H or t-butyl, f) R ２１ is H or benzyl, or g) The compound of claim 35, which is any combination of (a) to (g).

37. (a) R １６ , R １７ , R １８ , R １９ , R ２０ , and R ２１ is a protecting group, or (b) R １６ , R １７ , R １８ , R １９ , R ２０ , and R ２１ 37. The compound of claim 35 or 36, wherein is H.

38. The compound has the formula: 【Chemical formula 28】 36. The compound of claim 35,

39. (i) the compound is a solvate, or (ii) the compound is crystalline, and optionally a) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.8 to 6.1° ± 0.2 degrees in combination with one or more peaks selected from 6.7 to 7.1° and 8.8 to 9.0°, or b) The compound according to any one of claims 35 to 38, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2θ of 5.0 to 5.2° ± 0.2 degrees in combination with one or more peaks selected from 5.3 to 5.4°, 5.7 to 6.0°, 6.1 to 6.2°, 7.6 to 7.9°, and 8.7 to 9.1°.

40. 39. A method of synthesizing a polypeptide of SEQ ID NO: 1, comprising: (i) conjugating a compound of any one of claims 35 to 38 via the N-terminus of the compound to the C-terminus of a polypeptide of SEQ ID NO: 3 to form a polypeptide of SEQ ID NO: 10; and (ii) conjugating the polypeptide of SEQ ID NO: 10 via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO:

11.

41. A compound of formula (VI) or a salt, solvate, or hydrate thereof: 【Chemical 29】 [In the formula, R ２２ is H or a protecting group, and R ２３ is H or a protecting group, and R ２４ is H or a protecting group, and R ２５ is H or a protecting group, optionally each protecting group independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

42. a) R ２２ is H or Boc, b) R ２３ is H or tert-butyl, c) R ２４ is H or trityl, d) R ２５ is H or tert-butyl, or e) The compound of claim 41, which is any combination of (a) to (d).

43. (a) R ２２ , R ２３ , R ２４ , and R ２５ is H, or (b) R ２２ , R ２３ , R ２４ , and R ２５ 43. The compound of claim 41 or 42, wherein at least one of (a) and (b) is a protecting group, or (c) both (a) and (b).

44. The compound has the formula (VI-a): 【Chemistry 30】 42. The compound of claim 41, wherein:

45. (i) the compound is a solvate, optionally a solvate formed from pentyl acetate, a mixture comprising pentyl acetate, ethyl acetate, or a mixture comprising 2-methyltetrahydrofuran and t-amyl methyl ether, and optionally the mixture comprising pentyl acetate is selected from a mixture comprising pentyl acetate and t-butyl ethyl ether, a mixture comprising pentyl acetate and t-amyl methyl ether, or a mixture comprising pentyl acetate and heptane; (ii) the compound is a desolvate or a partially desolvate; or (iii) the compound is crystalline, optionally a) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 6.3 to 6.4° ±0.2 degrees in combination with one or more peaks selected from 4.5°, 7.1°, 13.0-13.1°, 15.9-16.0°, and 18.4-18.6°; or b) The compound of any one of claims 41 to 44, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.0 to 7.2° ±0.2 degrees in combination with one or more peaks selected from 5.0 to 5.4°, 7.6 to 7.7°, 8.8 to 8.9°, 9.4 to 9.5°, and 12.5 to 12.7°.

46. 42. A method of synthesizing a compound of claim 41, comprising: Formula (Y 保護 ) and a compound of formula (Aib 保護 ) to react with a compound of formula (Y 保護 -Aib 保護 ) forming a compound of formula (I) 【Chemical 31】 (In the formula, R ２２ , R ２３ , and R ２６ is a protecting group) - the protecting group R ２６ By removing 保護 -Aib), 【Chemical 32】 - the formula (Y 保護 -Aib) and a compound of formula (Q 保護２ ) to react with a compound of formula (Y 保護 -Aib-Q 保護２ ) forming a compound of formula (I) 【Chemical Formula 33】 (In the formula, R ２４ and R ２７ is a protecting group) - the protecting group R ２７ By removing 保護 -Aib-Q 保護１ ) forming a compound of formula (I) 【Chemical 34】 - the formula (Y 保護 -Aib-Q 保護１ ) and a compound of formula (G 保護 ) to form a compound of formula (VI), 【Chemical 35】 (In the formula, R ２５ is a protecting group), and optionally one or more protecting groups R ２２ , R ２３ , R ２４ , and R ２５ The method of claim 1, further comprising:

47. 47. The method of claim 46, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

48. 60. A method of synthesizing a polypeptide, wherein (i) the polypeptide is SEQ ID NO: 12, said method comprising conjugating a compound of any one of claims 56 to 59 via the C-terminus of said compound to the N-terminus of a polypeptide of SEQ ID NO: 13, or (ii) the polypeptide is SEQ ID NO: 14, said method comprising conjugating a compound of any one of claims 56 to 59 via the C-terminus of said compound to the N-terminus of a polypeptide of SEQ ID NO:

15.

49. A compound of formula (VII) or a salt, solvate, or hydrate thereof: 【Chemical 36】 [In the formula, R ２８ is H or a protecting group, and R ２９ is H or a protecting group, and R ３０ is H or a protecting group, optionally each protecting group independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

50. a) R ２８ is H or Boc, b) R ２９ is H or trityl, c) R ３０ is H or tert-butyl, or d) The compound of claim 49, which is any combination of (a) to (c).

51. (a) R ２８ , R ２９ , and R ３０ is H, or (b) R ２８ , R ２９ , and R ３０ 51. The compound of any one of claims 49-50, wherein at least one of (a) and (b) is a protecting group, or (c) both (a) and (b).

52. The compound has the formula (VII-a): 【Chemical 37】 50. The compound of claim 49, wherein:

53. (i) the compound is a solvate, optionally a solvate formed from any one of a mixture of acetonitrile and methyl tert-butyl ether, a mixture of nitromethane and methyl tert-butyl ether, a mixture of tetrahydrofuran and methyl tert-butyl ether, methyl acetate, and ethyl acetate, or (ii) the compound is crystalline, optionally a) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 4.8° ±0.2 degrees in combination with one or more peaks selected from 5.6°, 6.2°, 14.8°, and 15.6°; or b) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.3° ±0.2 degrees in combination with one or more peaks selected from 7.7°, 10.5°, 11.3°, 11.6°, and 14.4°; or c) The compound according to any one of claims 49 to 52, wherein the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 6.2° and 6.9° ±0.2 degrees.

54. 50. A method of synthesizing a compound of claim 49, comprising: -Formula (H(dnp) 保護 ) and a compound of formula (Aib 保護 ) to form a compound of formula (H(dnp) 保護 -Aib 保護 ) forming a compound of formula (I) 【Chemical Formula 38】 (In the formula, R ２８ and R ３１ is a protecting group) - the protecting group R ３１ By removing (H(dnp) 保護 -Aib), 【Chemical 39】 - the formula (H(dnp) 保護 -Aib) and a compound of formula (Q 保護２ ) to form a compound of formula (H(dnp) 保護 -Aib-Q 保護２ ) forming a compound of formula (I) 【Chemistry 40】 (In the formula, R ２９ and R ３２ is a protecting group) - the protecting group R ３２ By removing (H(dnp) 保護 -Aib-Q 保護１ ) forming a compound of formula (I) 【Chemistry 41】 - the formula (H(dnp) 保護 -Aib-Q 保護１ ) and a compound of formula (G 保護 ) to form a compound of formula (VII), 【Chemistry 42】 (In the formula, R ３０ is a protecting group), and optionally one or more protecting groups R ２８ , R ２９ , and R ３０ The method of claim 1, further comprising:

55. 55. The method of claim 54, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

56. 53. A method of synthesizing a polypeptide of SEQ ID NO: 16, comprising conjugating a compound of any one of claims 49 to 52 via the C-terminus of said compound to the N-terminus of a polypeptide of SEQ ID NO:

17.

57. A compound of formula (VIII) or a salt, solvate, or hydrate thereof: 【Chemistry 43】 [In the formula, R ３３ is H or a protecting group, and R ３４ is H or a protecting group, and R ３５ is H or a protecting group, optionally each protecting group independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

58. a) R ３３ is H or Boc, b) R ３４ is H or trityl, c) R ３５ is H or tert-butyl, or d) The compound of claim 57, which is any combination of (a) to (c).

59. (a) R ３３ , R ３４ , and R ３５ is H, or (b) R ３３ , R ３４ , and R ３５ 59. The compound of claim 57 or 58, wherein at least one of (a) and (b) is a protecting group, or (c) both (a) and (b).

60. The compound has the formula (VIII-a): 【Chemical 44】 58. The compound of claim 57, wherein:

61. (i) the compound is a solvate, optionally a solvate formed from any one of a mixture of tetrahydrofuran and methyl tert-butyl ether, a mixture of tetrahydrofuran and heptane, a mixture of 1,4-dioxane and water, a mixture of ethyl acetate and methyl tert-butyl ether, and a mixture of acetonitrile and methyl tert-butyl ether, or (ii) the compound is crystalline, optionally a) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 4.7° ±0.2 degrees in combination with one or more peaks selected from 5.5°, 8.2°, 10.1°, 11.8°, 13.3°, 13.6°, and 18.9°; or b) The compound of any one of claims 57 to 60, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.8° ± 0.2 degrees in combination with one or more peaks selected from 5.3°, 8.9°, 9.2°, 15.2°, 18.6°, and 19.5°.

62. 58. A method of synthesizing a compound of claim 57, comprising: -Formula (H(trt) 保護 ) and a compound of formula (Aib 保護 ) to form a compound of formula (H(trt) 保護 -Aib 保護 ) forming a compound of formula (I) 【Chemistry 45】 (In the formula, R ３３ and R ３６ is a protecting group) - the protecting group R ３６ By removing (H(trt) 保護 -Aib), 【Chemistry 46】 - The above formula (H(trt) 保護 -Aib) and a compound of formula (Q 保護２ ) to form a compound of formula (H(trt) 保護 -Aib-Q 保護２ ) forming a compound of formula (I) 【Chemistry 47】 (In the formula, R ３３ and R ３７ is a protecting group) - the protecting group R ３７ By removing (H(trt) 保護 -Aib-Q 保護１ ) forming a compound of formula (I) 【Chemistry 48】 - The above formula (H(trt) 保護 -Aib-Q 保護１ ) and a compound of formula (G 保護 to form a compound of formula (VIII), 【Chemistry 49】 (In the formula, R ３５ is a protecting group), and optionally one or more protecting groups R ３３ , R ３４ , and R ３５ The method of claim 1, further comprising:

63. 63. The method of claim 62, wherein each protecting group is independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

64. 61. A method of synthesizing a polypeptide of SEQ ID NO: 16, comprising conjugating a compound of any one of claims 57 to 60 via the C-terminus of said compound to the N-terminus of a polypeptide of SEQ ID NO:

17.

65. A compound of formula (IX) or a salt, solvate, or hydrate thereof: 【Chemistry 50】 [In the formula, R ３８ is H or a protecting group, and R ３９ is H or a protecting group, and R ４０ is H or a protecting group, and R ４１ is H or a protecting group, and R ４２ is H or a protecting group, and R ４３ is H or a protecting group, optionally each protecting group independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

66. a) R ３８ is H or Fmoc, b) R ３９ is H or tert-butyl, c) R ４０ is H or tert-butyl, d) R ４１ is H or tert-butyl, e) R ４２ is H or Boc, f) R ４３ is H or tert-butyl, or g) The compound of claim 65, which is any combination of (a) to (f).

67. (a) R ３８ , R ３９ , R ４０ , R ４１ , R ４２ , and R ４３ is H, or (b) R ３８ , R ３９ , R ４０ , R ４１ , R ４２ , and R ４３ 67. The compound of claim 65 or 66, wherein at least one of (a) and (b) is a protecting group, or (c) both (a) and (b).

68. The compound has the formula (IX-a): 【Chemistry 51】 66. The compound of claim 65, wherein:

69. (i) the compound is a solvate, optionally a solvate formed from any one of a mixture of methyl acetate and dibutyl ether, a mixture of acetone and dibutyl ether, a mixture of acetonitrile and dibutyl ether, a mixture of ethyl acetate and dibutyl ether, a mixture of methyl acetate and heptane, and a mixture of methyl ethyl ketone and dibutyl ether; (ii) the compound is a desolvate or anhydrous; or (iii) the compound is crystalline, optionally a) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.3° ±0.2 degrees in combination with one or more peaks selected from 6.0°, 6.9°, 7.2°, 8.0°, 12.2°, and 15.6°; or b) The compound according to any one of claims 65 to 68, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.8° ± 0.2 degrees in combination with one or more peaks selected from 4.4°, 6.6°, 10.1°, 11.4°, 13.4°, and 15.5°, or wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 4.5° and 5.5° ± 0.2 degrees in combination with one or more peaks selected from 6.0° and 7.3°.

70. 66. A method of synthesizing a compound of claim 65, comprising: -Formula (D 保護 ) and a compound of formula (Y 保護２ ) to react with a compound of formula (D 保護 -Y 保護２ ) forming a compound of formula (I) 【Chemistry 52】 (In the formula, R ３８ , R ３９ , R ４０ , and R ４４ is a protecting group) - the protecting group R ４４ By removing (D 保護 -Y 保護１ ) forming a compound of formula (I) 【Chemistry 53】 - the formula (D 保護 -Y 保護１ ) and a compound of formula (S 保護２ ) to react with a compound of formula (D 保護 -Y 保護１ -S 保護２ ) forming a compound of formula (I) 【Chemical 54】 (In the formula, R ４１ and R ４５ is a protecting group) - the protecting group R ４５ By removing (D 保護 -Y 保護１ -S 保護１ ) forming a compound of formula (I) 【Chemistry 55】 - the formula (D 保護 -Y 保護１ -S 保護１ ) and a compound of formula (K 保護２ ) to form a compound of formula (IX), 【Chemical 56】 (In the formula, R ４２ and R ４３ is a protecting group), and optionally one or more protecting groups R ３８ , R ３９ , R ４０ , R ４１ , R ４２ , and R ４３ The method of claim 1, further comprising:

71. 71. The method of claim 70, wherein the protecting group is selected from Boc, Fmoc, tert-butyl, and trityl groups.

72. 16. A method of synthesizing a polypeptide of SEQ ID NO: 16, comprising: (i) conjugating a compound of any one of claims 65-68 via the C-terminus of said compound to the N-terminus of a polypeptide of SEQ ID NO: 18; and (ii) conjugating a compound of claim 132 via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO:

19.

73. A compound of formula (X), or a salt, solvate, or hydrate thereof: 【Chemical 57】 [In the formula, R ４６ is H or a protecting group, and R ４７ is H or a protecting group, and R ４８ is H or a protecting group, and R ４９ is H or a protecting group, and R ５０ is H or a protecting group, and R ５１ is H or a protecting group, optionally each protecting group independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

74. a) R ４６ is H or tert-butyl, b) R ４７ is H or tert-butyl, c) R ４８ is H or Boc, d) R ４９ is H or Fmoc, e) R ５０ is H or tert-butyl, f) R ５１ is H or tert-butyl, or g) The compound of claim 73, which is any combination of (a) to (f).

75. (a) R ４６ , R ４７ , R ４８ , R ４９ , R ５０ , and R ５１ is H, or (b) R ４６ , R ４７ , R ４８ , R ４９ , R ５０ , and R ５１ 75. The compound of claim 73 or 74, wherein at least one of (a) and (b) is a protecting group, or (c) both (a) and (b).

76. The compound has the formula (X-a): 【Chemistry 58】 74. The compound of claim 73, wherein:

77. (i) the compound is a solvate, optionally a solvate formed from ethanol or isopropyl alcohol, (ii) the compound is a desolvate, or (iii) the compound is crystalline, optionally a) the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 18.1° and 18.7° ±0.2 degrees in combination with one or more peaks selected from 5.7°, 8.7°, 13.7°, 14.3°, 15.9°, and 16.2°; or b) the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 5.9° and 10.5° ±0.2 degrees in combination with one or more peaks selected from 7.1°, 8.9°, 14.6°, and 16.6°; or c) the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 7.8° and 20.3° ±0.2 degrees in combination with one or more peaks selected from 5.8°, 15.5°, and 19.5°; or d) The compound of any one of claims 73 to 76, wherein the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 5.9° and 7.4° ±0.2 degrees in combination with one or more peaks selected from 6.5°, 6.9°, and 14.8°.

78. 74. A method of synthesizing a compound of claim 73, comprising: Formula (Y 保護 ) and a compound of formula (S 保護２ ) to react with a compound of formula (Y 保護 -S 保護２ ) forming a compound of formula (I) 【Chemical Formula 59】 (In the formula, R ４６ , R ４７ , R ４８ , and R ５２ is a protecting group) - the protecting group R ５２ By removing 保護 -S 保護１ ) forming a compound of formula (I) 【Chemistry 60】 - the formula (Y 保護 -S 保護１ ) and a compound of formula (K 保護２ ) to react with a compound of formula (Y 保護 -S 保護１ -K 保護２ ) forming a compound of formula (I) 【Hua 61】 (In the formula, R ４９ and R ５３ is a protecting group), - the protecting group R ５３ By removing 保護 -S 保護１ -K 保護１ ) forming a compound of formula (I) 【Hua 62】 - the formula (Y 保護 -S 保護１ -K 保護１ ) and a compound of formula (Y 保護２ ) to form a compound of formula (X), 【Chemistry 63】 (In the formula, R ５０ and R ５１ is a protecting group), and optionally one or more protecting groups R ４６ , R ４７ , R ４８ , R ４９ , R ５０ , and R ５１ The method of claim 1, further comprising:

79. 79. The method of claim 78, wherein the protecting group is selected from Boc, Fmoc, tert-butyl, and trityl groups.

80. 16. A method of synthesizing a polypeptide of SEQ ID NO: 16, comprising: (i) conjugating a compound of any one of claims 73 to 76 via its C-terminus to the N-terminus of a polypeptide of SEQ ID NO: 20; and (ii) conjugating the resulting compound via its N-terminus to the C-terminus of a polypeptide of SEQ ID NO:

21.

81. A compound of formula (XI) or a salt, solvate, or hydrate thereof: 【Hua 64】 [In the formula, R ６４ is H or a protecting group, and R ６５ is H or a protecting group, and R ６６ is H or a protecting group, optionally each protecting group independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

82. a) (a) R ６４ , R ６５ , and R ６６ is H, or b) (b) R ６４ , R ６５ , and R ６６ is a protecting group, or c) The compound of claim 81, which is both (c)(a) and (b).

83. The compound has the formula: 【Chemistry 65】 82. The compound of claim 81,

84. 84. The compound of any one of claims 81 to 83, wherein (i) the compound is a solvate, or (ii) the compound is crystalline, optionally in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 6.1° and 8.5° ±0.2 degrees in combination with one or more peaks selected from 5.8°, 16.9°, 18.5°, 18.8°, 19.3°, and 20.9°.

85. A compound of formula (XII) or a salt, solvate, or hydrate thereof: 【Hua 66】 [In the formula, R ６１ is H or a protecting group, and R ６２ is H or a protecting group, and R ６３ is H or a protecting group, optionally each protecting group independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

86. a) R ６１ is H or tert-butyl; b) R ６２ is H or tert-butyl; c) R ６３ is H or tert-butyl; d) The compound of claim 85, which is any combination of (a) to (c).

87. (a) R ６１ , R ６２ , and R ６３ is H, or (b) R ６１ , R ６２ , and R ６３ 87. The compound of claim 85 or 86, wherein at least one of (a) and (b) is a protecting group, or (c) both (a) and (b).

88. The compound has the formula (XII-a): 【Chemical Formula 67】 86. The compound of claim 85, wherein:

89. (i) the compound is a solvate, (ii) the compound is a desolvate or anhydrous, or (iii) the compound is crystalline, and optionally a) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 11.4° ±0.2 degrees in combination with one or more peaks selected from 6.0°, 8.9°, 12.7°, 13.6°, 14.6°, 17.0°, and 18.8°; or b) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 10.6° ±0.2 degrees in combination with one or more peaks selected from 7.1°, 12.1°, 13.6°, 14.2°, 15.2°, 16.0°, and 16.8°; or c) The compound of any one of claims 85 to 88, wherein the compound is in the form of a crystalline solid characterized by peaks in an X-ray powder diffraction pattern at diffraction angles 2-theta of 10.1° and 15.5° ±0.2 degrees in combination with one or more peaks selected from 6.1°, 8.7°, 11.4°, 16.6°, and 19.2°.

90. A compound of formula (XIII) or a salt, solvate, or hydrate thereof: 【Chemistry 68】 [In the formula, R ６７ is H or a protecting group, and R ６８ is H or a protecting group, and R ６９ is H or a protecting group, optionally each protecting group independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

91. a) R ６７ is H or tert-butyl; b) R ６８ is H or tert-butyl; c) R ６９ is H or tert-butyl; d) The compound of claim 90, which is any combination of (a) to (c).

92. (a) R ６７ , R ６８ , and R ６９ is H, or (b) R ６７ , R ６８ , and R ６９ 92. The compound of claim 90 or 91, wherein at least one of (a) and (b) is a protecting group, or (c) both (a) and (b).

93. The compound has the formula: 【Chemical Formula 69】 91. The compound of claim 90,

94. (i) the compound is a solvate, (ii) the compound is a desolvate or anhydrous, or (iii) the compound is crystalline, and optionally a) the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 5.0° ±0.2 degrees in combination with one or more peaks selected from 8.3, 9.7, and 11.2°; or b) The compound according to any one of claims 90 to 93, wherein the compound is in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.2° ± 0.2 degrees in combination with one or more peaks selected from 5.3, 8.1, 14.4, and 16.2°.

95. A compound of formula (XIV) or a salt, solvate, or hydrate thereof: 【Chemistry 70】 [In the formula, R ７０ is H or a protecting group, and R ７１ is H or a protecting group, and R ７２ is H or a protecting group, and R ７３ is H or a protecting group, and R ７４ is H or a protecting group, optionally each protecting group independently selected from Boc, Fmoc, tert-butyl, and trityl groups.

96. a) R ７０ is H, Fmoc, or tert-butyl; b) R ７１ is H or tert-butyl; c) R ７２ is H or tert-butyl; d) R ７３ is H or tert-butyl; e) R ７４ is H, Fmoc, or tert-butyl; f) The compound of claim 95, which is any combination of (a) to (e).

97. (a) R ７０ , R ７１ , R ７２ , R ７３ , and R ７４ is H, or (b) R ７０ , R ７１ , R ７２ , R ７３ , and R ７４ 97. The compound of claim 95 or 96, wherein at least one of (a) and (b) is a protecting group, or (c) both (a) and (b).

98. The compound has the formula: 【Chemical 71】 96. The compound of claim 95,

99. 99. The compound of any one of claims 95 to 98, wherein (i) the compound is a solvate, (ii) the compound is a desolvate or anhydrous, or (iii) the compound is crystalline, optionally in the form of a crystalline solid characterized by a peak in an X-ray powder diffraction pattern at a diffraction angle 2-theta of 7.5° ± 0.2 degrees in combination with one or more peaks selected from 6.1, 8.7, 10.6, 15.0, 16.1, and 18.6°.

Images