Process and compounds for preparing spiroligomers

Abstract

The present invention provides novel intermediates and other compounds prepared using the compounds of formula I. These novel compounds include spiroligomers that are of special importance in the peptide industry. The processes described herein provide products that are produced in high purity and high yields and are therefore highly efficient.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to and the benefit of U.S. Provisional Patent Application No. 63 / 307,869, filed February 8, 2022, the disclosure of which is incorporated by reference in its entirety herein.

[0002] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was made by Government support under DE-EE0008321 awarded by the Department of Energy and HDTRA1-16-1-0047 awarded by the Defense Threat Reduction Agency. The Government has certain rights in this invention.

[0003] Technical Field The present disclosure is directed to processes and compounds for preparing spiroligomers. [Background technology]

[0004] 2. Background of the Invention Bis-peptides, also known as spiroligomers, are ladder oligomers or polymers formed from amino acids. Spiroligomers are useful as therapeutics, catalysts, sensors, antibody mimetics, and nanotechnology. Their shape can be tailored based on the stereochemistry and sequence of the monomers. In synthetic bis-peptides, a diketopiperazine ring is formed using amino acids to create the spiroligomer.

[0005] The standard procedure for synthesizing spiroligomers is via a stepwise approach by sequentially adding a single bis-amino acid at each stage of the synthesis. Although this type of process allows for stereochemical control, they are relatively inefficient. For example, such processes typically yield less than a few milligrams of spiroligomer, and the steps are difficult to carry out, and the process is often time-consuming and labor-intensive.

[0006] Thus, there is a need in the art for an effective method for preparing spiroligomers through simple steps and / or in high yields. The present invention addresses this unmet need in the art. Summary of the Invention

[0007] In some embodiments, the present invention provides a method for the preparation of a pharmaceutical composition comprising the steps of: 1 X, R 1 C(O)H or R 1 C(O)R 2 and a compound of formula I: [ka] {where, R 1 , R 2 , X, R 4 , x, and z are defined herein} with a compound of formula II: [ka] wherein y is an integer 0 or 1; provided that the compound of formula I is 1 When reacting with X, y is an integer 0 and R 2 wherein x is the absence of a reducing agent; for a time and under conditions effective to produce

[0008] In other aspects, the present invention provides compounds, such as spiroligomers, produced using the processes described herein. In a further aspect, the present invention provides compounds of formula I, III, IV, VII, and XV: [ka] {where, R 1 , R 2 , R 4 , R 5 , x, y, and z are defined herein. The present invention provides a process for preparing

[0009] In yet another aspect, the present invention provides a compound of formula II: [ka] {where, R 1 , R 2 , R 5 and y is as defined herein} or a pharma- ceutically acceptable salt thereof.

[0010] In yet another aspect, the present invention provides a method for producing a composition comprising the steps of: [ka] and the like. Other aspects and embodiments of the present invention will become apparent and readily apparent from the following detailed description of the invention.

[0011] The following detailed description of various embodiments of the invention will be better understood when read in conjunction with the accompanying drawings. For the purpose of illustrating the invention, there are shown in the drawings exemplary embodiments. It should be understood, however, that the invention is not limited to the precise arrangements and instrumentalities of the embodiments shown in the drawings. [Brief description of the drawings]

[0012] [Figure 1] FIG. 1 shows a schematic diagram for the fluorenylmethoxycarbonyl (Fmoc) protection of functionalized bis-amino acid building blocks using a temporary Cu complexation strategy, together with an efficient multikilogram-scale synthesis of bis-amino acid precursors. [Diagram 2] FIG. 2 shows a schematic diagram for the synthesis of the Cbz building block. [Diagram 3] FIG. 3 shows a schematic diagram for the synthesis of Fmoc building blocks. [Figure 4] FIG. 4 shows a schematic diagram for the synthesis of functionalized building blocks and Pfp esters. [Diagram 5]FIG. 5 shows schematic diagrams of the chemical structures of spiroligomers (a) T2, (b) T3, (c) T4, and (d) the modeled structure of T1 based on ROESY correlation with CANDO and GAFF energy minimization. [Figure 6] FIG. 6 shows a schematic diagram for the synthesis of functionalized spiroligomer T1 with selected ROESY correlations and modeled structure. [Figure 7] FIG. 7 shows a schematic diagram and a corresponding representative 1H NMR spectrum for compound S2a, previously reported (Levins, CG et al., Journal of the American Chemical Society 2003, 125 (16), 4702-4703). [Figure 8] FIG. 8 shows a schematic diagram and the corresponding representative 1H NMR spectrum for compound S3a, previously reported (Levins, CG et al., Journal of the American Chemical Society 2003, 125 (16), 4702-4703). [Figure 9] FIG. 9 shows a schematic diagram and the corresponding representative 1H NMR spectrum for compound 2a, as previously reported (Levins, CG et al., Journal of the American Chemical Society 2003, 125 (16), 4702-4703). [Figure 10] FIG. 10 shows a schematic diagram and a corresponding representative 1H NMR spectrum for compound S2b, previously reported (Levins, CG et al., Journal of the American Chemical Society 2003, 125 (16), 4702-4703). [Figure 11] FIG. 11 shows a schematic diagram and a corresponding representative 1H NMR spectrum for compound S3b, previously reported (Levins, CG et al., Journal of the American Chemical Society 2003, 125 (16), 4702-4703). [Figure 12] FIG. 12 shows a schematic diagram and the corresponding representative 1H NMR spectrum for compound 2b, as previously reported (Levins, CG et al., Journal of the American Chemical Society 2003, 125 (16), 4702-4703). [Figure 13] FIG. 13 shows a schematic diagram and corresponding representative 1H NMR and 13C NMR spectra for compound 4a (3S,5S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-amino-5-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid. [Figure 14] FIG. 14 shows a schematic diagram and corresponding representative 1H NMR and 13C NMR spectra for compound 4b (3R,5S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-amino-5-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid. [Figure 15] FIG. 15 shows a schematic diagram and corresponding representative 1H NMR and 13C NMR spectra for compound 4c (3R,5R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-amino-5-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid. [Figure 16] FIG. 16 shows a schematic diagram and corresponding representative 1H NMR and 13C NMR spectra for compound 4d (3R,5S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-amino-5-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid. [Figure 17] FIG. 17 shows a schematic diagram of compound 5a (3S,5S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-5-(tert-butoxycarbonyl)-3-((naphthalen-2-ylmethyl)amino)pyrrolidine-3-carboxylic acid and the corresponding representative 1H NMR and 13C NMR spectra. [Figure 18]FIG. 18 shows a schematic diagram of bisamino acid 5b (3R,5S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-5-(tert-butoxycarbonyl)-3-(isopentylamino)pyrrolidine-3-carboxylic acid and corresponding representative 1H NMR and 13C NMR spectra. [Figure 19] FIG. 19 shows a schematic diagram and corresponding representative 1H NMR and 13C NMR spectra for compound 5c (3R,5R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-5-(tert-butoxycarbonyl)-3-((pyridin-4-ylmethyl)amino)pyrrolidine-3-carboxylic acid. [Figure 20] FIG. 20 shows a schematic diagram and corresponding representative 1H NMR and 13C NMR spectra for compound 5d (3S,5R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-5-(tert-butoxycarbonyl)-3-((3,4-dichlorobenzyl)amino)pyrrolidine-3-carboxylic acid. [Figure 21] FIG. 21 shows a schematic diagram and corresponding representative 1H NMR and 13C NMR spectra for compound 6a 1-((9H-fluoren-9-yl)methyl) 2-(tert-butyl) 4-(perfluorophenyl)(2S,4S)-4-((naphthalen-2-ylmethyl)amino)pyrrolidine-1,2,4-tricarboxylate. [Figure 22] FIG. 22 shows a schematic diagram and corresponding representative 1H NMR and 13C NMR spectra for compound 6b 1-((9H-fluoren-9-yl)methyl) 2-(tert-butyl) 4-(perfluorophenyl)(2S,4R)-4-(isopentylamino)pyrrolidine-1,2,4-tricarboxylate. [Figure 23]FIG. 23 shows a schematic diagram and corresponding representative 1H NMR and 13C NMR spectra for compound 6c 1-((9H-fluoren-9-yl)methyl) 2-(tert-butyl) 4-(perfluorophenyl)(2R,4R)-4-((pyridin-4-ylmethyl)amino)pyrrolidine-1,2,4-tricarboxylate. [Figure 24] FIG. 24 shows a schematic diagram and corresponding representative 1H NMR and 13C NMR spectra for compound 6d 1-((9H-fluoren-9-yl)methyl) 2-(tert-butyl) 4-(perfluorophenyl)(2R,4S)-4-((3,4-dichlorobenzyl)amino)pyrrolidine-1,2,4-tricarboxylate. [Diagram 25] FIG. 25 shows representative images of Fmoc building block synthesis during (a) hydrogenolysis of Cbz building block using Pd / C, (b) Fmoc protection using Fmoc-Cl, (c) Pd / C removal using a Celite column, (d) Cu removal using EDTA solution and product precipitation, and (d) product isolation by vacuum filtration. [Figure 26] FIG. 26 shows a representative HPLC chromatograph of crude T1 at 220 nm without purification. [Figure 27] FIG. 27 shows representative QTOF LCMS results of T1: (top) LC chromatogram at 220 nm; (middle) total ion current chromatogram; (bottom) mass spectrogram, with theoretical and observed monoisotopic peaks in red and black, respectively. [Figure 28] FIG. 28 shows representative QTOF LCMS results of T2: (top) LC chromatogram at 220 nm; (middle) total ion current chromatogram; (bottom) mass spectrogram, with theoretical and observed monoisotopic peaks in red and black, respectively. [Figure 29]FIG. 29 shows representative QTOF LCMS results of T3: (top) LC chromatogram at 220 nm; (middle) total ion current chromatogram; (bottom) mass spectrogram, with theoretical and observed monoisotopic peaks in red and black, respectively. [Diagram 30] FIG. 30 shows representative QTOF LCMS results of T4: (top) LC chromatogram at 220 nm; (middle) total ion current chromatogram; (bottom) mass spectrogram, with theoretical and observed monoisotopic peaks in red and black, respectively. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] Detailed Description of the Invention The present invention provides an efficient process for preparing spiroligomers. In contrast to processes in the art, the process described herein allows the preparation of a large number of spiroligomers with highly preconfigured structures. By doing so, various building blocks of various length sequences can be assembled, resulting in an almost limitless number of highly preconfigured molecules with programmable shapes and functional group displays.

[0014] The present invention employs key and unique steps that provide the ability to prepare spiroligomers and their intermediates in high yields. Initially, the process allowed for the preparation of intermediates in 40-50 gram batches, which far exceeded the yields described in the art. After refining the process, multi-kilogram batches of intermediates and products could be prepared.

[0015] Among other features, one unique step includes the use of the delicate Fmoc group to protect amines at key positions of the molecule. Various protecting groups have been used in the process to prepare spiroligomers. However, the inventors found that simply exchanging out the traditional protecting group for the Fmoc group was not effective and further reduced the already low yield of spiroligomers to a much lower yield. The Fmoc protecting group can also be easily functionalized by simple steps with a wide range of functional groups. Another key step includes the use of metals such as copper for selective complexation of amino acid groups to allow selective protection of other amines with the Fmoc group. These key steps resulted in high yields, i.e., >90%, and purity of more than 95%.

[0016] Finally, the spiroligomers, once formed, could be successfully stringently bound using non-stringent conditions for a minimal period of time.

[0017] definition As used herein, unless expressly indicated otherwise, the singular forms "a," "an," and "the" include plural references and a reference to a particular numerical value includes at least that particular value. Thus, for example, a reference to "a material" is a reference to at least one such material and equivalents thereof known to those skilled in the art.

[0018] When values ​​are expressed as approximations by use of the descriptor "about," it is understood that the particular value forms another embodiment. In general, use of the term "about" indicates an approximation that may vary depending on the desired properties sought to be obtained by the disclosed subject matter and should be interpreted in the particular context in which it is used based on its function. In some aspects of the present invention, "about" refers to a range of values ​​that is ±10% of the recited value. For example, "about 10" refers to "9 to 11," as well as "10." One of ordinary skill in the art can interpret this as routine. In some cases, the number of significant figures used for a particular value may be one non-limiting way of determining the extent of the term "about." In other cases, the incremental changes used in a series of values ​​may be used to determine the intended range available to the term "about" for each value. When present, all ranges are inclusive and combinable. That is, reference to values ​​described in ranges includes all values ​​within that range. When a parameter range is provided, it is understood that all integers within that range, as well as their tenths, are also provided by the present invention. For example, "20 to 40 mg" includes 20.0 mg, 20.1 mg, 20.2 mg, 20.3 mg, etc., up to a maximum of 40.0 mg.

[0019] When lists are presented, it is to be understood that each individual member of that list and every combination of that list is to be construed as a separate embodiment unless otherwise specified. For example, a list of embodiments presented as "A, B, or C" is to be construed as including the embodiments "A," "B," "C," "A or B," "A or C," "B or C," or "A, B, or C."

[0020] It is understood that certain features of the invention that are described herein for clarity in the context of separate embodiments may also be provided in combination within a single embodiment. That is, each individual embodiment is considered to be compatible with any other embodiment, and such combinations are considered to be separate embodiments, unless expressly incompatible or excluded. Conversely, different features of the invention that are described in the context of a single embodiment for ease of description may also be provided separately or in any subcombination. It is further noted that the claims may be drafted to exclude any optional element. Thus, this description is intended to serve as a basis for prescriptive limitations to the use of exclusive language such as "only", "only" and the like in connection with the recitation of claim elements, or the use of a "negative" limitation. Finally, although an embodiment may be described as part of a series of steps or as part of a more general structure, each step may be considered an independent embodiment in itself.

[0021] The term "halo," as used herein, refers to Cl, F, Br, or I. In some embodiments, halo is Cl. In other embodiments, halo is F. In further embodiments, halo is Br. In yet other embodiments, halo is I. "C0" as used herein refers to the absence of a carbon atom. For example, C 0-6 Alkyl OH is -OH and C 1-6 Refers to alkyl.

[0022] The term "alkyl" as used herein refers to an aliphatic hydrocarbon containing 1 to 12 carbon atoms, i.e., C 1-12 In some embodiments, alkyl is methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In other embodiments, alkyl is C 1-8 In a further embodiment, alkyl is C 1-6 In yet further embodiments, alkyl is C 1-4In yet other embodiments, alkyl is C 1-4 It is an alkyl. Examples of alkyl groups include methyl, ethyl, propyl (e.g., n-propyl, isopropyl), butyl (e.g., sec-butyl, tert-butyl, isobutyl), 3-pentyl, hexyl, heptyl, octyl, nonyl, and decyl. The alkyl may be unsubstituted or substituted, i.e., optionally substituted, as described herein. In certain embodiments, the alkyl is substituted with two substituents. In further embodiments, the alkyl is substituted with one substituent. In other embodiments, the alkyl is substituted with three substituents. In yet further embodiments, the alkyl is unsubstituted.

[0023] The term "alkenyl" as used herein refers to an alkyl containing one or more carbon-carbon double bonds. In some embodiments, an alkenyl group contains one, two, or three carbon-carbon double bonds. In other embodiments, an alkenyl contains one carbon-carbon double bond. In further embodiments, an alkenyl is a C 2-6 In yet another embodiment, the alkenyl is C 2-4 In still further embodiments, the alkenyl is C 3-4 Alkenyl. Non-limiting exemplary alkenyl groups include ethenyl, propenyl, isopropenyl, butenyl, sec-butenyl, pentenyl, hexenyl, and -CH=C(CH3)2. Alkenyl can be unsubstituted or substituted. In some embodiments, alkenyl is substituted with two substituents. In further embodiments, alkenyl is substituted with one substituent. In yet other embodiments, alkenyl is substituted with three substituents. In even further embodiments, alkenyl is unsubstituted.

[0024] The term "alkynyl" as used herein refers to an alkyl group containing one or more carbon-carbon triple bonds. In some embodiments, an alkynyl group contains one, two, or three carbon-carbon triple bonds. In other embodiments, an alkynyl group contains one carbon-carbon triple bond. In further embodiments, an alkynyl group is a C 2-6 In yet another embodiment, alkynyl is C 2-4 In still further embodiments, alkynyl is C 3-4 Alkynyl. Non-limiting exemplary alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, hexynyl, among others. Alkynyl can be unsubstituted or substituted. In some embodiments, alkynyl is substituted with two substituents. In further embodiments, alkynyl is substituted with one substituent. In yet other embodiments, alkynyl is substituted with three substituents. In even further embodiments, alkynyl is unsubstituted.

[0025] The term "alkoxy" as used herein refers to an optionally substituted alkyl as defined herein, containing an oxygen atom within the group. In some embodiments, the alkoxy group is an alkyl attached to a terminal oxygen atom. In other embodiments, the alkoxy group is a C 1-6 In a further embodiment, the alkoxy group is C 1-4 Examples of alkoxy groups include methoxy (OCH3), ethoxy (OCH2CH3 or CH2OCH3), propoxy (e.g., -O n Pr, -O i Pr), or butoxy (e.g., -O n Bu, -O i Bu, -O s Bu, -O tBu). The alkoxy can be unsubstituted or substituted. In some embodiments, the alkoxy is substituted with two substituents. In further embodiments, the alkoxy is substituted with one substituent. In yet other embodiments, the alkoxy is substituted with three substituents. In even further embodiments, the alkoxy is unsubstituted.

[0026] The term "cycloalkyl" as used herein refers to a saturated or partially unsaturated cyclic aliphatic hydrocarbon containing 1 to 3 rings and 3 to 12 carbon atoms, i.e., C 3-12 Cycloalkyl or C 3-12 In some embodiments, the cycloalkyl has two rings. In other embodiments, the cycloalkyl has one ring. In further embodiments, the cycloalkyl is saturated. In yet other embodiments, the cycloalkyl has one or two double bonds. In still further embodiments, the cycloalkyl is C 3-8 In yet a further embodiment, cycloalkyl is C 3-7 In yet other embodiments, cycloalkyl is C 3-7 In another embodiment, the cycloalkyl is C 3-6 In a further embodiment, cycloalkyl is C 3-6 Cycloalkyl. Examples of cycloalkyl groups include, among others, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, norbornyl, decalin, adamantyl, cyclohexenyl, and cyclopentenyl. Cycloalkyl can be unsubstituted or substituted. In some embodiments, cycloalkyl is substituted with two substituents. In further embodiments, cycloalkyl is substituted with one substituent. In yet other embodiments, cycloalkyl is substituted with three substituents. In yet further embodiments, cycloalkyl is unsubstituted.

[0027] The term "cyanoalkyl" as used herein refers to an alkyl group, as described, that is substituted with one or more CN. In some embodiments, a cyanoalkyl contains one CN substituent. In other embodiments, a cyanoalkyl contains two CN substituents. 1-6 Examples of cyanoalkyl include, but are not limited to, CH2CN, CH2CH2CN, CHCNCH3.

[0028] The term "haloalkyl" as used herein refers to an alkyl group, as described, substituted with one or more halos. In some embodiments, the haloalkyl contains one or more F, i.e., it is also a fluoroalkyl. In other embodiments, the haloalkyl contains one halo (e.g., F). In further embodiments, the haloalkyl contains two halos (e.g., F). In yet other embodiments, the haloalkyl contains three halos (e.g., F). 1-6 Examples of haloalkyl include, but are not limited to, CH2F, CHF2, CF3, CH2CFH2, CH2CF2H, CH2CH2CF3, among others.

[0029] The term "haloalkoxy" as used herein refers to an alkoxy group, as described, substituted with one or more halo. In some embodiments, the haloalkoxy contains one or more F, i.e., it is also a fluoroalkyl. In other embodiments, the haloalkoxy contains one halo (e.g., F). In further embodiments, the haloalkoxy contains two halos (e.g., F). In yet other embodiments, the haloalkoxy contains three halos (e.g., F). 1-6 Examples of haloalkoxy include, but are not limited to, OCH2F, OCHF2, OCF3, OCH2CFH2, OCH2CF2H, OCH2CH2CF3, among others.

[0030] The term "aryl" as used herein refers to a monocyclic or bicyclic unsaturated ring system having 5 to 14 carbon atoms, i.e., C 5-14 In some embodiments, aryl has 6 to 12 carbon atoms, i.e., C 6-12 In a further embodiment, the aryl has 6 to 10 carbon atoms, i.e., C 6-10 In another embodiment, the aryl has 6 to 8 carbon atoms, i.e., C 6-8 It is aryl. Examples of aryl groups include, but are not limited to, phenyl, naphthyl (1-naphthyl, 2-naphthyl), phenanthryl, anthracyl, indenyl, azulenyl, biphenyl, biphenylenyl, and fluorenyl groups. In some embodiments, the aryl group is phenyl or naphthyl. The term "aryl" also encompasses a phenyl group fused to a cycloalkyl. The aryl may be unsubstituted or substituted. In some embodiments, the aryl is an optionally substituted phenyl.

[0031] The term "heteroaryl" as used herein refers to a monocyclic or bicyclic aromatic ring system having 5-14 ring atoms, i.e., 5-14 membered heteroaryl. Heteroaryl contains carbon atoms and one or more heteroatoms, i.e., oxygen, nitrogen, and sulfur. In some embodiments, heteroaryl contains 1, 2, 3, or 4 oxygen, nitrogen, and / or sulfur. In other embodiments, heteroaryl contains 3 heteroatoms. In further embodiments, heteroaryl contains 2 heteroatoms. In still other embodiments, heteroaryl contains 1 heteroatom. In still other embodiments, heteroaryl is 5-10 membered heteroaryl. In still further embodiments, heteroaryl is 5-6 membered heteroaryl. In other embodiments, heteroaryl is 5 membered. In further embodiments, heteroaryl is 6 membered. Examples of heteroaryl groups include thienyl (e.g., thien-2-yl, thien-3-yl), benzo[b]thienyl, naphtho[2,3-b]thienyl, thianthrenyl, furyl (e.g., 2-furyl, 3-furyl, 4-furyl), benzofuryl, pyranyl, thiophenyl, benzofuranyl, isobenzofuranyl, benzoxazolyl, chromenyl, xanthenyl, pyrrolyl (e.g., pyrrol-2-yl, pyrrol-3-yl), imidazolyl (e.g., imidazol-2-yl, imidazol-4-yl, 1-methylimidazolyl), pyrazolyl (e.g., pyrazol-3-yl, pyrazol-4-yl, pyrazol-5-yl), pyrazolyl, ... Lysyl (e.g., pyridin-1-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl), pyrazinyl (e.g., pyrazin-2-yl, pyrazin-3-yl, pyrazin-5-yl, pyrazin-6-yl), pyrimidinyl (e.g., pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl), pyridazinyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl (e.g., 1,8-naphthyridinyl), cinnolinyl, triazolyl (e.g., 1,2,4-triazol-1-yl, 1,2,4-triazol-3-yl, 1,2,4-triazol-5-yl), thiadiazolyl (e.g., 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl), oxadiazolyl (e.g., 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl), quinazolinyl, pteridinyl, 4H-carbazolyl, carbazolyl, β-carbolinyl, phenanthridinyl, acridinyl, phenanthrolinyl, phenazinyl, thiazolyl thiazolyl (e.g., thiazol-2-yl, thiazol-4-yl, thiazol-5-yl, 2-methylthiazolyl), isothiazolyl (e.g., isothiazol-3-yl (isothiazol-4-yl, isothiazol-5-yl), phenothiazolyl, isoxazolyl (e.g., isoxazol-3-yl, isoxazol-4-yl, isoxazol-5-yl, 3,5-dimethylisoxazolyl), furazanyl, pyrazolo[1,5-a]pyridinyl, benzisothiazolyl, imidazole[1,5-a]pyridinyl (e.g., imidazole pyrrolo[1,5-a]pyridin-1-yl), pyrrolo[1,2]pyridazinyl (e.g., pyrrolo[2]pyridazin-5-yl, pyrrolo[1,2]pyridazin-6-yl), benzo[d]thiazolyl (e.g., benzo[d]thiazol-3-yl, benzo[d]thiazol-2-yl), benzo[d]imidazolyl (e.g., benzo[d]imidazol-2-yl), benzo[d]oxazolyl (e.g., benzo[d]oxazol-2-yl), benzo[c]isoxazolyl (e.g., benzo[c]isoxazol-3-yl), isothiazolyl, 4, Examples of the aryl group include 5,6,7-tetrahydropyrazolo[1,5-a]pyridinyl, phenoxazinyl, triazinyl, indolizinyl, quinolinyl, 1,2,3,4-tetrahydroquinolinyl, isoquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl, benzo[b]thiophenyl, 1H-indazolyl (e.g., indazol-3-yl), benzthiazolyl, 4H-quinolizinyl, quinoxalinyl, phenothiazinyl, oxazolyl (e.g., oxazol-2-yl, oxazol-4-yl, oxazol-5-yl), and 2-pyrenyl.

[0032] The term "heteroaryl" also includes N-oxides. Heteroaryl may be unsubstituted or substituted. In some embodiments, heteroaryl is substituted with two substituents. In further embodiments, heteroaryl is substituted with one substituent. In still other embodiments, heteroaryl is substituted with three substituents. In even further embodiments, heteroaryl is unsubstituted. Substitution can occur at any available carbon or heteroatom (e.g., nitrogen), or both, as permitted by the valence of the substituents. Heteroaryl also includes heteroaryl groups having a fused, optionally substituted cycloalkyl or a fused, optionally substituted heterocyclyl.

[0033] The term "heterocyclyl" as used herein refers to a non-aromatic saturated or partially unsaturated group containing one, two, or three rings having 3 to 14 ring members, i.e., 3- to 14-membered heterocyclyl. Heterocyclyl groups contain carbon atoms and one or more oxygen, sulfur, and / or nitrogen atoms, which can be oxidized or quaternized. In some embodiments, the ring CH2- is replaced with -C(=O)-. The term "heterocyclyl" also encompasses groups having a fused, optionally substituted aryl group, such as indolinyl or chroman-4-yl. In some embodiments, a heterocyclyl group is C 4-6Heterocyclyl. Heterocyclyl can be optionally linked to the rest of the molecule by any available carbon or heteroatom. Examples of heterocyclyl include azetidinyl (e.g., azetidin-1-yl, azetidin-2-yl, azetidin-3-yl), dioxanyl, tetrahydropyranyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, pyrrolidinyl, indolinyl, oxetanyl, tetrahydrofuranyl, tetrahydrothiophenyl, 2-pyrazolinyl, pyrazolidinyl, trithianyl, indolizinyl, benzo[b]thiophenyl, 1H-indazolyl, benzthiazolyl, 4H-quinolizinyl, quinolinyl, 1,2,3,4-tetrahydroquinolinyl, isoquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl, quinoxalinyl, and phenothiazinyl. The heterocyclyl may be unsubstituted or substituted. In some embodiments, the heterocyclyl is substituted with two substituents. In further embodiments, the heterocyclyl is substituted with one substituent. In yet other embodiments, the heterocyclyl is substituted with three substituents. In even further embodiments, the heterocyclyl is unsubstituted.

[0034] Any of the groups / substituents identified above or herein may be one or more of CN, halo, NO, OH, NH, C 1-6 Alkoxy, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, NH(C 1-6 alkyl), N(C 1-6 Alkyl)(C 1-6 Alkyl), Aryl, O-Aryl, C 1-6 Alkyl SH, -C(O)C 1-6 Alkyl, -C(O)OC 1-6 Alkyl, -C(O)aryl, -C(O)Oaryl, -C(O)heteroaryl, -C(O)Oheteroaryl, -C(O)heterocyclyl, -C(O)Oheterocyclyl, C 1-6 Cyanoalkyl, C 3-8 Cycloalkyl, C 1-6 Haloalkoxy, C1-6 Haloalkyl, heteroaryl, heterocyclyl, -C 1-6 -Alkyl(heterocyclyl), -C 1-6 -Alkyl(aryl), -C 1-6 -Alkyl(heteroaryl), -C 1-6 -Alkyl (cycloalkyl), C 1-6 It may be substituted with hydroxyalkyl, C(O)NH2, or OC(O)NH2.

[0035] The term "nucleobase" as used herein refers to an optionally substituted adenine, cytosine, guanine, thymine, or uracil. In some embodiments, the nucleobase is a purine base, such as adenine or guanine. In other embodiments, the nucleobase is a pyrimidine base, such as cytosine, uracil, or thymine. In further embodiments, the nucleobase is a modified nucleobase, such as a modified purine nucleobase or a modified pyrimidine nucleobase. Examples of modified nucleobases include hypoxanthine, xanthine, 7-methylguanine, inosine, xanthosine, 7-methylguanosine, 5,6-dihydrouracil, 5-methylcytosine, 5-hydroxymethylcytosine, dihydrouridine, or 5-methylcytidine. In yet other embodiments, the nucleobase is an artificial nucleobase, such as isoguanine, isocytosine, 2-amino-6-(2-thienyl)purine, or pyrrole-2-carbaldehyde.

[0036] The term "amino acid" as used herein refers to a naturally occurring or non-naturally occurring amino acid. In some embodiments, the amino acid is a naturally occurring amino acid. In certain aspects, the amino acid is arginine (Arg), histidine (His), lysine (Lys), aspartate (Asp), glutamate (Glu), serine (Ser), threonine (Thr), asparagine (Asn), glutamine (Gln), cysteine ​​(Cys), selenocysteine ​​(Sec), glycine (Gly), proline (Pro), alanine (Ala), valine (Val), isoleucine (Ile), leucine (Leu), methionine (Met), phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp), or pyrrolysine. In further embodiments, the amino acid is a non-naturally occurring amino acid. In certain embodiments, the amino acid is 4-aminobenzoic acid (PABA), alloisoleucine, allothreonine, carboxyglutamic acid, cystathionine, D-alanine, dehydroalanine, D-glutamate, diaminopimelic acid, dienkolic acid, glycine betaine, homocysteine, homonorleucine, homoserine, hydroxyglycine, hydroxyproline, hypusine, isoserine, isovaline, lanthionine, N-ethylalanine, N-ethylglycine, N-ethyl β-alanine, N-isopropylglycine, N-methylalanine, N-methyl β-alanine, norleucine, norvaline, N-propyl Glycine, O-methyl-homoserine, ornithine, pipecolic acid, pyroglutamic acid, sarcosine, selenocysteine, selenohomocysteine, selenomethionine, selenoethionine, taurine, t-leucine, α,β-diaminopropionic acid, α,γ-diaminobutyric acid, α-aminoisobutyric acid, α-amino-n-butyric acid, α-amino-n-heptanoic acid, α-hydroxy-γ-aminobutyric acid, β-alanine, β-aminoisobutyric acid, β-amino-n-butyric acid, γ-aminobutyric acid, δ-aminolevulinic acid, 1-aminocyclopropane-1-carboxylic acid, azetidine-2-carboxylic acid, cycloleucine, or pseudoproline. In some embodiments, the amino acid is protected by a suitable protecting group as described herein.

[0037] The term "sugar" as used herein refers to a monosaccharide or oligosaccharide as known in the art. In some embodiments, the sugar is a monosaccharide, such as glucose, fructose, or galactose. In other embodiments, the sugar is a disaccharide or a disaccharide (two monosaccharides linked by a glycosidic bond), such as sucrose, lactose, or maltose. In some embodiments, the sugar is protected by a suitable protecting group as described herein.

[0038] Ranges: Throughout the present invention, various aspects of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Thus, the description of a range should be considered to have all the specifically disclosed possible subranges as well as individual numerical values ​​within that range. For example, the description of a range such as 1-6 should be considered to have the specifically disclosed subranges such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, etc. as well as the individual numbers within that range, e.g., 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

[0039] process The present invention relates, in part, to a compound of formula II: [ka] The present invention relates to a process for preparing

[0040] In compounds of formula II, y is the integer 0 or 1. In some embodiments, y is the integer 0. In other embodiments, y is the integer 1. In these compounds, x is the integer 1 or 2. In some embodiments, x is the integer 1. In other embodiments, x is the integer 2. Similarly, z is the integer 1 or 2. In some embodiments, z is the integer 1. In other embodiments, z is the integer 2. In further embodiments, x and z are not both 2.

[0041] R 1 is optionally replaced by C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 In some embodiments, R is an optionally substituted aryl. 1 is optionally replaced by C 1-6 Alkyl, such as methyl, ethyl, propyl, butyl, pentyl, or hexyl. In other embodiments, R 1 is optionally replaced by C 2-6 alkenyl, such as ethenyl, propenyl, butenyl, pentenyl, or hexenyl. In a further embodiment, R 1 is optionally replaced by C 2-6 Alkynyl, such as ethynyl, propynyl, butynyl, pentynyl, or hexynyl. In yet another embodiment, R 1 is optionally substituted aryl, such as phenyl. 2 is absent, H, or optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 In some embodiments, R is an optionally substituted aryl. 2 is absent. 2 is H. In a further embodiment, R 2 is optionally replaced by C 1-6 Alkyl, such as methyl, ethyl, propyl, butyl, pentyl, or hexyl. In other embodiments, R 2 is optionally replaced by C 2-6 alkenyl, such as ethenyl, propenyl, butenyl, pentenyl, or hexenyl. In a further embodiment, R 2 is optionally replaced by C 2-6Alkynyl, such as ethynyl, propynyl, butynyl, pentynyl, or hexynyl. In yet another embodiment, R 2 is optionally substituted aryl, such as phenyl. 1 and R 2 may be substituted with one or more groups. In some embodiments, R 1 and / or R 2 is independently substituted with 1, 2, 3, 4, or 5 groups. In some embodiments, R 1 and R 2 When is substituted, the substituents may be the same or different.

[0042] In certain embodiments, R 1 and R 2 is C 0-6 AlkylOH, C 0-6 Alkyl SH, C 0-6 Alkyl NH2, C 0-6 Alkyl-OC 0-6 Alkyl, C 0-6 Alkyl-SC 0-6 Alkyl, C 0-6 AlkylC(O)OH, C 0-6 AlkylC(O)(C 1-6 Alkyl), C 0-6 AlkylC(O)O(C 1-6 Alkyl), C 0-6 AlkylN3,C 0-6 AlkylC(O)NH2, C 0-6 Alkyl-C(O)N(C 1-6 alkyl)OH, optionally substituted C 3-7 Cycloalkyl, optionally substituted C 5-7 Cycloalkenyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, -NHC(NH2)(=N(C 0-6 alkyl), -NHC(NH2)(=S(C 0-6 alkyl), nucleobase, or amino acid, wherein any carbon atom of said alkyl, alkenyl, or alkynyl is independently substituted with a heteroatom, i.e., O, S, SO, SO2, or NR7 is optionally replaced by

[0043] In other embodiments, R 1 and R 2 C such as OH or CH2OH 0-6 In a further embodiment, R 1 and R 2 is C such as SH and CH2SH. 0-6 In yet another embodiment, R 1 and R 2 is NH2, methanamine, ethanamine, propanamine, N,N-dimethylmethanamine, dimethylamine, or N,N,N-trimethylmethanamine, etc. 0-6 In still further embodiments, R 1 and R 2 -OC 1-6 Alkyl or C such as OCH3 0-6 Alkyl-OC 0-6 In another embodiment, R 1 and R 2 -SC 1-6 Alkyl, etc., or C, such as -SCH3 0-6 Alkyl-SC 0-6 In a further embodiment, R 1 and R 2 is C(O)OH, ethanoic acid, acetic acid, or propionic acid, etc. 0-6 In yet another embodiment, R 1 and R 2 is -C(O)(C 1-6 alkyl), -CH2-C(O)(C 1-6 alkyl), or -C(O)CH3, etc. 0-6 AlkylC(O)(C 1-6 In still further embodiments, R 1 and R 2 C such as methyl formate or methyl acetate 0-6AlkylC(O)O(C 1-6 In another embodiment, R 1 and R 2 is N3 or -CH2N3 0-6 In a further embodiment, R 1 and R 2 is C(O)NH2, ethanoamide, or propionamide, etc. 0-6 In yet another embodiment, R 1 and R 2 is -C(O)N(C 1-6 C(O)N(CH3)OH, carboxyhydroxamide, ethanohydroxamide, or propionohydroxamide, etc. 0-6 Alkyl-C(O)N(C 1-6 In still further embodiments, R 1 and R 2 is an optionally substituted C such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc., or cyclopropyl, etc., or cyclobutyl, etc. 3-7 In another embodiment, R 1 and R 2 is an optionally substituted C such as cyclopentenyl, cyclohexenyl, or cycloheptenyl. 5-7 In a further embodiment, R 1 and R 2 is independently an optionally substituted aryl, such as phenyl, p-cresol, 1-methoxy-benzyl, naphthyl, 4-methyl-phenol, or 1-methoxy-4-methyl-benzene, or phenyl. 1 and R 2is independently optionally substituted heteroaryl, such as imidazolyl, 2-pyrenyl, 1-methylimidazolyl, indolyl, pyridinyl, such as 2-pyridinyl, 3-pyridinyl, or 4-pyridinyl, triazolyl, or imidazolyl. 1 and R 2 is independently an optionally substituted heterocyclyl, such as azetidinyl, dioxanyl, tetrahydropyranyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, pyrrolidinyl, indolinyl, oxetanyl, tetrahydrofuranyl, tetrahydrothiophenyl, 2-pyrazolinyl, pyrazolidinyl, trithianyl, indolizinyl, benzo[b]thiophenyl, 1H-indazolyl, benzthiazolyl, 4H-quinolizinyl, quinolinyl, 1,2,3,4-tetrahydroquinolinyl, isoquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl, quinoxalinyl, phenothiazinyl, and the like. 1 and R 2 are independently -NHC(NH2)(=N(C) such as guanidinyl, methyl-guanidinyl, ethyl-guanidinyl, or propyl-guanidinyl. 0-6 In a further embodiment, R 1 and R 2 are independently -NHC(NH2)(=S(C 0-6 In some embodiments, R 1 and R 2 may be protected with a removable protecting group. For example, an active nitrogen can be protected using an Alloc group.

[0044] In yet another embodiment, R 1 and R 2 is independently a nucleobase. Examples of such nucleobases include, but are not limited to, guaninyl, adeninyl, cytosinyl, thyminyl, or any combination thereof.

[0045] In yet another embodiment, R1 and R 2 are independently an amino acid, a sugar, or any combination thereof. In some embodiments, the amino acid is a β-amino acid. Examples of amino acids include, but are not limited to, arginine (Arg), histidine (His), lysine (Lys), aspartate (Asp), glutamic acid (Glu), serine (Ser), threonine (Thr), asparagine (Asn), glutamine (Gln), cysteine ​​(Cys), selenocysteine ​​(Sec), glycine (Gly), proline (Pro), alanine (Ala), valine (Val), isoleucine (Ile), leucine (Leu), methionine (Met), phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp), pyrrolysine, or any combination thereof.

[0046] In a further embodiment, R 1 and R 2 are linked together with carbon, they are joined together to form C groups such as cyclopropyl, cyclobutyl, or cyclopentyl. 3-8 In some embodiments, R 1 and R 2 are linked together to form a cyclopropyl. 1 and R 2 are linked together to form a cyclobutyl. In a further embodiment, R 1 and R 2 are linked together to form cyclopentyl. 1 and R 2 are linked together to form cyclohexyl.

[0047] In certain embodiments, any carbon atom within an alkyl, alkenyl, or alkynyl group can be O, S, SO, SO, or NR 7In some embodiments, any carbon atom of the alkyl, alkenyl, or alkynyl group is optionally substituted with an O heteroatom. In further embodiments, any carbon atom of the alkyl, alkenyl, or alkynyl group is optionally substituted with an S heteroatom. In still other embodiments, any carbon atom of the alkyl, alkenyl, or alkynyl group is optionally substituted with an SO group. In even further embodiments, any carbon atom of the alkyl, alkenyl, or alkynyl group is optionally substituted with an SO group. In other embodiments, any carbon atom of the alkyl, alkenyl, or alkynyl group is optionally substituted with an NH heteroatom, or N(C 1-4 alkyl), or NR, such as NCH 7 is optionally replaced by

[0048] R 7 , H, C 1-4 Alkyl, C 3-4 Alkenyl, C 3-4 Alkynyl, or C 1-4 a bridged alkyl, where a bridge is formed between a nitrogen and a carbon atom of said heteroatom-containing chain to form a ring, where said ring is optionally selected from the group consisting of Ar 1 In some embodiments, R 7 is H. In other embodiments, R 7 is C such as CH3 1-4 In a further embodiment, R 7 is C 3-4 In yet another embodiment, R 7 is C 3-4 In still further embodiments, R 7 is C 1-4 a bridged alkyl, where a bridge is formed between a nitrogen and a carbon atom of said heteroatom-containing chain to form a ring, where said ring is optionally selected from the group consisting of Ar 1 are merged into.

[0049] Ar 1 is C3-6 In some embodiments, Ar is cycloalkyl, aryl, heterocyclyl, or heteroaryl. 1 is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, etc. 3-6 In another embodiment, Ar is cycloalkyl. 1 is aryl, such as phenyl, 1-naphthyl, 2-naphthyl, indenyl, azulenyl, fluorenyl, or anthracyl. 1 is heterocyclyl, such as azetidinyl, aziridinyl, piperidinyl, or azepanyl. 1 is heteroaryl such as acridinyl, benzimidazolyl, benzthiazolyl, benzo[b]furanyl, benzo[b]thiophenyl, carbazolyl, cinnolinyl, furyl such as 2-furyl or 3-furyl, imidazolyl, indolyl such as 1H-indazolyl, indolizinyl, 3H-indolyl, indolinyl, isoindolyl, isoquinolinyl, isoxazolyl, 1,8-naphthyridinyl, oxadiazolyl such as 1,2,3-oxadiazolyl, oxazolyl, phenazinyl, phenothiazinyl, phenoxazinyl, pterid ... quinolinyl, pyrazinyl, pyrazolyl, 2-pyrazolinyl, pyrazolidinyl, pyridazinyl, pyrimidinyl, pyridyl such as 2-pyridyl, 3-pyridyl or 4-pyridyl, pyrrolyl, 4H-quinolidinyl, quinolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 1,2,3,4-tetrahydroquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl, 1,3,4-thiadiazolyl, thiazolyl, 1,2,3-triazolyl, 1,3,5-triazinyl, 1,3,5-trithianyl, or thienyl such as 2-thienyl or 3-thienyl.

[0050] Ar 1 is one or more H, halo, OH, NO2, -SO3H, CF3, OCF3, CH3, etc. 1-6 Alkyl, C 2-6 Alkenyl, OCH3, etc. 1-6 Alkoxy, OC 3-4Alkenyl, -O-benzyl, -O-phenyl, 1,2-methylenedioxy, -NR 5 R 6 , -C(O)OH, -C(O)(C 1-6 alkyl), C(O)O(C 1-6 alkyl), -C(O)NH(C 1-6 alkyl), -a carboxamide, -C(O)NH(C 3-5 alkenyl), -C(O)N(C 1-6 Alkyl)(C 1-6 alkyl), -C(O)(C 3-5 Alkenyl)(C 3-6 alkenyl), morpholinyl, piperidinyl, -O-Ar 2 , -CH2-(CH2) q -Ar 2 , -O-(CH2) q -Ar 2 , -(CH2) q -O-Ar 2 or -CH=CH-Ar 2 Optionally replaced by R 5 and R 6 are independently H, C 1-6 Alkyl, C 3-6 Alkenyl, C 3-6 alkynyl, or benzyl. 2 is 4-methoxyphenyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, pyrazyl, quinolyl, 3,5-dimethylisoxazolyl, 2-methylthiazolyl, thiazolyl, 2-thienyl, 3-thienyl or pyrimidinyl. q is an integer of 0 to 2.

[0051] In the compound of formula II, X is a leaving group. One of skill in the art would be able to readily determine a suitable leaving group for use as "X." In some embodiments, X is a halo or sulfonate. In other embodiments, X is a halo, such as chloro, fluoro, or bromo. In further embodiments, X is a sulfonate, such as p-toluenesulfonate (OTs), methanesulfonate (OMs), or trifluoromethanesulfonate (OTf). R in the compound of formula II 4is a protecting group. In some embodiments, R 4 is C 1-6 alkyl or 4-{N-[1-(4,4-dimethyl-2,6-dioxocyclohexylidene)-3-methylbutyl]amino}benzyl ester (DMab). 4 C such as t-butyl 1-6 In certain embodiments, the compound of formula II has the structure of formula II-A: [ka] .

[0052] In a further embodiment, the compound of formula II has the structure of formula II-A. In another embodiment, the compound of formula II has the structure of formula II-B. In a further embodiment, the compound of formula II has the structure of formula II-C.

[0053] The compound of formula II can be reacted with a compound of formula I, as follows: [ka] R 1 X, R 1 C(O)H or R 1 C(O)R 2 It is prepared by contacting with

[0054] In some embodiments, the compound of formula I is 1 When reacting with X, y is an integer 0 and R 2 is non-existent.

[0055] The organic solvent used to prepare the compound of formula II can be selected by one skilled in the art. In some embodiments, the organic solvent is a polar organic solvent. In other embodiments, the organic solvent is an alcoholic solvent such as methanol; tetrahydrofuran, or an ether solvent such as ethyl acetate. The reducing agent used to form the compound of formula II can be selected by one skilled in the art. In some embodiments, the reducing agent is NaBH3, NaBH3CN, or Na(CH3COO)3BH. In other embodiments, the reducing agent is NaBH3CN.

[0056] In some embodiments, the compound of formula I has the structure of formula IA: [ka] .

[0057] In a further embodiment, the compound of formula I has the structure of formula IA. In another embodiment, the compound of formula I has the structure of formula IB. In a further embodiment, the compound of formula I has the structure of formula IC.

[0058] The compound of formula I may be reacted with a compound of formula III, as follows, for a time and under conditions effective to produce the compound of formula I: [ka] wherein M is a transition metal having a +2 oxidation state, with a chelating agent. In some embodiments, M is vanadium, manganese, iron, cobalt, nickel, copper, or zinc. In other embodiments, M is copper. In further embodiments, M is manganese. In still other embodiments, M is iron. In even further embodiments, M is cobalt, and in other embodiments, M is nickel. In further embodiments, M is zinc.

[0059] The chelating agent used to prepare the compound of formula I can be selected by one of skill in the art. In some embodiments, the chelating agent is ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), or hydroxyethylethylenediaminetriacetic acid (HEDTA), or a salt thereof. In further embodiments, the chelating agent is Na2EDTA·H2O or CaNa2EDTA. In other embodiments, the chelating agent is H2S, a thiazolidinethione, glycine, and / or other chelating agents.

[0060] The organic solvent for preparing the compound of formula I can be selected by one of skill in the art. In some embodiments, the organic solvent is a polar organic solvent. In other embodiments, the organic solvent is an ether solvent, a chlorinated organic solvent, or a combination thereof. In further embodiments, the organic solvent is an ether solvent, such as methyl t-butyl ether. In yet other embodiments, the organic solvent is chloroform, which is a chlorinated organic solvent. In even further embodiments, the organic solvent is a mixture of methyl t-butyl ether and chloroform. When a combination / mixture of solvents is utilized, the ratio is about 1:1 to about 1:5. In some embodiments, the ratio of methyl t-butyl ether to chloroform is about 1:1 to about 1:5.

[0061] In some embodiments, the compound has the structure of Formula III-A, III-B, III-C, III-D, III-E, III-F, or III-G: [ka] [ka] [ka]

[0062] In further embodiments, the compound of formula III has the structure of formula III-A. In other embodiments, the compound of formula III has the structure of formula III-B. In further embodiments, the compound of formula III has the structure of formula III-C. In yet other embodiments, the compound of formula III has the structure of formula III-D. In still further embodiments, the compound of formula III has the structure of formula III-E. In other embodiments, the compound of formula III has the structure of formula III-F. In still further embodiments, the compound of formula III has the structure of formula III-G.

[0063] The compound of formula III may be reacted with a compound of formula IV, for a time and under conditions effective to produce a compound of formula IV: [ka] with 9-fluorenylmethoxycarbonyl chloride. The organic solvent utilized to prepare the compound of formula IV is a polar organic solvent. In some embodiments, the organic solvent is ethyl acetate or any solvent of similar polarity that allows the reaction to occur. In some embodiments, the organic solvent allows the reaction to occur. In some aspects, the compound of formula IV has the structure of formula IV-A, IV-B, IV-C, IV-D, IV-E, IV-F, or IV-G: [ka] .

[0064] In a further embodiment, the compound of formula IV has the structure of formula IV-A. In another embodiment, the compound of formula IV has the structure of formula IV-B. In a further embodiment, the compound of formula IV has the structure of formula IV-C. In yet another embodiment, the compound of formula IV has the structure of formula IV-D. In an even further embodiment, the compound of formula IV has the structure of formula IV-E. In another embodiment, the compound of formula IV has the structure of formula IV-F. In a further embodiment, the compound of formula IV has the structure of formula IV-G.

[0065] The compound of formula IV may be reacted with a compound of formula V, for a time and under conditions effective to produce the compound of formula IV: [ka] with a metal(II) source. In some embodiments, the metal(II) source is a Ni(II) source or a copper(II) source. In other embodiments, the metal source is a copper(II) source, such as a copper(II) salt. In further embodiments, the metal(II) source is a Ni(II) source. In yet other embodiments, a copper(II) salt, such as copper chloride or copper sulfate. The solvent can be selected by one of skill in the art. In some embodiments, the solvent is water. In other embodiments, the solvent is a mixture of water, optionally with one or more organic co-solvents.

[0066] In some embodiments, the compound of formula V has the structure of formula VA, VB, or VC: [ka] .

[0067] In further embodiments, the compound of formula V has the structure of formula VA. In other embodiments, the compound of formula V has the structure of formula VB. In further embodiments, the compound of formula V has the structure of formula VC.

[0068] The compound of formula II can be reacted with an activating agent and a base to provide an intermediate, a compound of formula VI-A, VI-B, or VI-C: [ka] may be contacted with

[0069] In certain embodiments, the compound of formula II is contacted with a compound of formula VI-A. In other embodiments, the compound of formula II is contacted with a compound of formula VI-B. In further embodiments, the compound of formula II is contacted with a compound of formula VI-C.

[0070] In some embodiments, AA is an optionally substituted amino acid. The amino acid can be selected by one skilled in the art. In some aspects, AA is a natural or non-natural amino acid. In certain aspects, AA is arginine (Arg), histidine (His), lysine (Lys), aspartate (Asp), glutamic acid (Glu), serine (Ser), threonine (Thr), asparagine (Asn), glutamine (Gln), cysteine ​​(Cys), selenocysteine ​​(Sec), glycine (Gly), proline (Pro), alanine (Ala), valine (Val), isoleucine (Ile), leucine (Leu), methionine (Met), phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp), or pyrrolysine. In certain aspects, AA is glycine. In other aspects, AA is alanine. In a further embodiment, AA is valine. In yet another embodiment, AA is leucine. In yet another embodiment, AA is isoleucine. In another embodiment, AA is proline. In yet another embodiment, AA is serine. In yet another embodiment, AA is threonine. In yet another embodiment, AA is asparagine. In another embodiment, AA is glutamine. In yet another embodiment, AA is cysteine. In yet another embodiment, AA is methionine. In yet another embodiment, AA is phenylalanine. In another embodiment, AA is tyrosine. In yet another embodiment, AA is tryptophan. In yet another embodiment, AA is aspartate. In yet another embodiment, AA is glutamic acid. In another embodiment, AA is histidine. In yet another embodiment, AA is lysine. In yet another embodiment, AA is arginine. In yet another embodiment, AA is selenocysteine. In yet another embodiment, AA is pyrrolysine. In a further embodiment, AA is β-alanine.

[0071] AA is also found in the following compounds: PABA, alloisoleucine, allothreonine, carboxyglutamic acid, cystathionine, D-alanine, dehydroalanine, D-glutamate, diaminopimelic acid, dhenkol acid, glycine betaine, homocysteine, homonorleucine, homoserine, hydroxyglycine, hydroxyproline, hypusine, isoserine, isovaline, lanthionine, N-ethylalanine, N-ethylglycine, N-ethyl β-alanine, N-isopropylglycine, N-methylalanine, N-methyl β-alanine, norleucine, norvaline, N-propylglycine, O-methyl-homoserine, ornithine , pipecolic acid, pyroglutamic acid, sarcosine, selenocysteine, selenohomocysteine, selenomethionine, selenoethionine, taurine, t-leucine, α,β-diaminopropionic acid, α,γ-diaminobutyric acid, α-aminoisobutyric acid, α-amino-n-butyric acid, α-amino-n-heptanoic acid, α-hydroxy-γ-aminobutyric acid, β-alanine, β-aminoisobutyric acid, β-amino-n-butyric acid, γ-aminobutyric acid, δ-aminolevulinic acid, 1-aminocyclopropane-1-carboxylic acid, azetidine-2-carboxylic acid, cycloleucine, or unnatural amino acids such as pseudoproline. In some embodiments, AA is PABA. In other embodiments, AA is alloisoleucine. In further embodiments, AA is allothreonine. In yet other embodiments, AA is carboxylglutamic acid. In still further embodiments, AA is cystathionine. In other embodiments, AA is D-alanine. In still further embodiments, AA is dehydroalanine. In still further embodiments, AA is D-glutamate. In still further embodiments, AA is diaminopimelic acid. In still further embodiments, AA is jenkolic acid. In other embodiments, AA is glycine betaine homocysteine. In still further embodiments, AA is homonorleucine. In still further embodiments, AA is homoserine. In still further embodiments, AA is hydroxyglycine. In other embodiments, AA is hydroxyproline. In still further embodiments, AA is hypusine. In still other embodiments, AA is isoserine.In still further embodiments, AA is isovaline. In other embodiments, AA is lanthionine. In still further embodiments, AA is N-ethylalanine. In still further embodiments, AA is N-ethylglycine. In still further embodiments, AA is N-ethyl β-alanine. In other embodiments, AA is N-isopropylglycine. In still further embodiments, AA is N-methylalanine. In still further embodiments, AA is N-methyl β-alanine. In still further embodiments, AA is norleucine. In other embodiments, AA is norvaline. In still further embodiments, AA is N-propylglycine. In still further embodiments, AA is O-methyl-homoserine. In still further embodiments, AA is ornithine. In other embodiments, AA is pipecolic acid. In still further embodiments, AA is pyroglutamic acid. In still further embodiments, AA is sarcosine. In still further embodiments, AA is selenocysteine. In other embodiments, AA is selenohomocysteine. In a further embodiment, AA is selenomethionine. In yet another embodiment, AA is selenoethionine. In an even further embodiment, AA is taurine. In another embodiment, AA is t-leucine. In a further embodiment, AA is α,β-diaminopropionic acid. In yet another embodiment, AA is α,γ-diaminobutyric acid. In an even further embodiment, AA is α-aminoisobutyric acid. In another embodiment, AA is α-amino-n-butyric acid. In a further embodiment, AA is α-amino-n-heptanoic acid. In a still further embodiment, AA is α-hydroxy-γ-aminobutyric acid. In an even further embodiment, AA is β-alanine. In another embodiment, AA is β-aminoisobutyric acid. In a further embodiment, AA is β-amino-n-butyric acid. In a still further embodiment, AA is γ-aminobutyric acid. In an even further embodiment, AA is δ-aminolevulinic acid. In another embodiment, AA is 1-aminocyclopropane-1-carboxylic acid. In a further embodiment, AA is azetidine-2-carboxylic acid. In yet another embodiment, AA is cycloleucine.In an even further embodiment, AA is pseudoproline. 4 The amino acid is attached to an oxygen atom bonded to the amino acid backbone. The amino acid can be bonded to the oxygen atom through any position of the amino acid backbone. For example, in some embodiments, the amino acid is attached to an oxygen atom through the carbonyl of the amino acid.

[0072] In these structures, R 5 is a resin. Suitable resins for use herein can be selected by one of skill in the art. In some embodiments, the resin contains a carboxy group. Preferably, the resin comprises a cleavable group. The term "cleavable group" as used herein refers to a group that is displaced from a compound of formula VC. In particular, the cleavable group is displaced from a compound of formula VC so that AA can be attached. In some embodiments, the cleavable group is OH or carboxyl. In other embodiments, the cleavable group is OH. In further embodiments, the cleavable group is carboxyl. In yet other embodiments, the cleavable group is a polystyrene resin.

[0073] The activating agent utilized may be selected by one of skill in the art. In some embodiments, the activating agent is selected from the group consisting of 1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (COMU), 1H-1,2,3-benzotriazol-1-yloxy-tris(pyrrolidino)-phosphonium hexafluorophosphate (PyBOP), 1-hydroxy-7-azabenzotriazole (HOAt), 2-(1H-7-azabenzotriazol-1-yloxy)dimethylamino-morpholino-carbenium ... N,N,N',N'-tetramethylformamidinium hexafluorophosphate, dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), ethyl cyano(hydroxyimino)acetate-O2 ]tri-1-pyrrolidinylphosphonium hexafluorophosphate (PyOxim), ethyl cyanohydroxyiminoacetate (Oxyma), N,N'-diisopropylcarbodiimide (DIC), N-hydroxybenzotriazole (HOBT), O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU), O-1,1,3,3-tetramethyluronium hexafluorophosphate (HCTU), or O-benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), or combinations thereof. In some embodiments, the activator is COMU. In other embodiments, the activator is PyBOP. In further embodiments, the activator is HOAt. In still other embodiments, the activator is HATU. In even further embodiments, the activator is BOP. In other embodiments, the activator is chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate, in further embodiments, the activator is DCC. In still other embodiments, the activator is DIC. In even further embodiments, the activator is PyOxim. In other embodiments, the activator is Oxyma. In further embodiments, the activator is DIC. In still other embodiments, the activator is HOBT. In even further embodiments, the activator is TBTU. In other embodiments, the activator is HCTU. In further embodiments, the activator is HBTU.

[0074] The solvent for this transformation can be selected by one skilled in the art. Preferably, the solvent is a polar solvent. Examples of solvents include NMP, DMF, DMSO, DCM, or mixtures thereof. In some embodiments, the solvent is NMP. In other embodiments, the solvent is DMF. In further embodiments, the solvent is DMSO. In yet other embodiments, the solvent is DCM.

[0075] The intermediate is then reacted with a compound of formula VII: [ka] with a deprotecting agent for a time and under conditions effective to produce

[0076] In some embodiments, the intermediate is a compound of formula VII-A, VII-B, or VII-C: [ka] is converted to

[0077] In some embodiments, the intermediate is converted to a compound of formula VII-A. In other embodiments, the intermediate is converted to a compound of formula VII-B. In further embodiments, the intermediate is converted to a compound of formula VII-C.

[0078] Those skilled in the art will be able to select a suitable deprotecting agent. In some embodiments, the deprotecting agent is a base. Examples of bases include, but are not limited to, piperidine, 4-methylpiperidine, piperazine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), or morpholine, or combinations thereof. In certain embodiments, the base is piperidine. In other embodiments, the base is 4-methylpiperidine. In further embodiments, the base is piperazine. In yet other embodiments, the base is DBU. In even further embodiments, the base is morpholine.

[0079] The compound of formula VII is then reacted with a compound of formula VIIIA, VIII-B, or VIII-C: [ka] is converted to

[0080] In some embodiments, the compound of formula VII-A is converted to a compound of formula VIII-A. In other embodiments, the compound of formula VII-B is converted to a compound of formula VIII-B. In further embodiments, the compound of formula VII-C is converted to a compound of formula VIII-C.

[0081] Such conversion is carried out by contacting a compound of formula VII with a compound of formula II in the presence of an activating agent, as described above, followed by contacting the reactants with a deprotecting agent, as described above, for a time and under conditions effective to produce a compound of formula VIII-A, VIII-B, or VIII-C.

[0082] Advantageously, the compound of formula VIII-A, VIII-B or VIII-C can be further reacted with a compound of formula II to give, for example, a compound of formula XV-A, XV-B or XV-C: [ka] It is also possible to provide corresponding larger compounds such as

[0083] In another embodiment, compound XV-A may be prepared. In a further embodiment, compound XV-B may be prepared. In yet another embodiment, compound XV-C may be prepared.

[0084] As a general principle, these larger compounds are formed by reacting the product of each reaction involving a compound of formula II with another compound of formula II. The number of times ("w") the reaction with a compound of formula II is carried out is determined by the size of the compound desired. In certain embodiments, the product is reacted with a compound of formula II 1-20 times (i.e., w is an integer from 1-20). In other embodiments, w is an integer 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. In further embodiments, w is an integer from 2 to 18, 2 to 16, 2 to 14, 2 to 12, 2 to 10, 2 to 8, 2 to 6, 2 to 4, 4 to 20, 4 to 18, 4 to 16, 4 to 14, 4 to 12, 4 to 10, 4 to 8, 4 to 6, 6 to 20, 6 to 18, 6 to 16, 6 to 14, 6 to 12, 6 to 10, 6 to 8, 8 to 20, 8 to 18, 8 to 16, 8 to 14, 8 to 12, 8 to 10, 10 to 20, 10 to 18, 10 to 16, 10 to 14, 10 to 12, 12 to 20, 12 to 18, 12 to 16, 12 to 14, 14 to 20, 14 to 18, 14 to 16, 16 to 20, 16 to 18, or 18 to 20. In still other embodiments, w is an integer from 1 to 8. In even further embodiments, w is an integer from 6 to 8.

[0085] The compound of formula VIII-A, VIII-B, or VIII-C is then reacted with a compound of formula IX-B, IX-B, or IX-C: [ka] may be converted to

[0086] In some embodiments, a compound of formula XIII-A is converted to a compound of formula IX-A. In other embodiments, a compound of formula VIII-B is converted to a compound of formula IX-B. In further embodiments, a compound of formula VIII-C is converted to a compound of formula IX-C. Such conversion is carried out by contacting a compound of formula VIII-A, VIII-B, or VIII-C with a compound of formula II in the presence of an activating agent as described above. The reaction is then contacted with a deprotecting agent, as described above, for a time and under conditions effective to produce a compound of formula IX-A, IX-B, or IX-C.

[0087] The compound of formula IX-A, IX-B, or IX-C is then reacted with a compound of formula XB, XB, or XC: [ka] may be converted to

[0088] In some embodiments, the compound of formula IX-A is converted to a compound of formula XA. In other embodiments, the compound of formula XIII-A is converted to a compound of formula XB. In further embodiments, the compound of formula XIII-A is converted to a compound of formula XC.

[0089] Such conversion is carried out by contacting a compound of formula IX-A, IX-B, or IX-C with a compound of formula II in the presence of an activating agent, as described above, and then contacting the reactants with a deprotecting agent, as described above, for a time and under conditions effective to produce a compound of formula XA, XB, or XC.

[0090] The compound of formula XA, XB, or XC is then reacted with a compound of formula XI-B, XI-B, or XI-C: [ka] may be converted to

[0091] In some embodiments, the compound of formula XA is converted to a compound of formula XI-A. In other embodiments, the compound of formula XB is converted to a compound of formula XI-B. In further embodiments, the compound of formula XC is converted to a compound of formula XI-C.

[0092] Compounds of formula XA, XB, or XC are converted to compounds of formula XI-A, XI-B, or XI-C by reaction with an activating agent as described herein and YZ. Y is defined as an aminocarbonyl group, and Z is a leaving group. The term "aminocarbonyl group" as used herein refers to any chemical functional group containing NH2 and C(O) groups. Y is H or an amino acid. In some embodiments, Y is H. In other embodiments, Y is an amino acid. Examples of amino acids include those described herein. The amino acid can be selected by one of skill in the art. In some embodiments, Y is a natural or non-natural amino acid. In certain embodiments, Y is Arg, His, Lys, Asp, Glu, Ser, Thr, Asn, Gln, Cys, Sec, Gly, Pro, Ala, Val, Ile, Leu, Met, Phe, Tyr, Trp, or pyrrolysine. In certain embodiments, Y is Gly. In other embodiments, Y is Ala. In a further embodiment, Y is Val. In yet another embodiment, Y is Leu. In yet another embodiment, Y is Ile. In another embodiment, Y is Pro. In a further embodiment, Y is Ser. In yet another embodiment, Y is Thr. In a still further embodiment, Y is Asp. In another embodiment, Y is Gln. In a further embodiment, Y is Cys. In yet another embodiment, Y is Met. In a still further embodiment, Y is Phe. In another embodiment, Y is Tyr. In a still further embodiment, Y is Trp. In a still further embodiment, Y is Asp. In a still further embodiment, Y is Glu. In another embodiment, Y is His. In a still further embodiment, Y is Lys. In a still further embodiment, Y is Arg. In a still further embodiment, Y is selenocysteine. In another embodiment, Y is pyrrolysine. In a still further embodiment, Y is β-alanine.

[0093] Y is also found in: PABA, alloisoleucine, allothreonine, carboxyglutamic acid, cystathionine, D-alanine, dehydroalanine, D-glutamate, diaminopimelic acid, djenkolic acid, glycine betaine, homocysteine, homonorleucine, homoserine, hydroxyglycine, hydroxyproline, hypusine, isoserine, isovaline, lanthionine, N-ethylalanine, N-ethylglycine, N-ethyl β-alanine, N-isopropylglycine, N-methylalanine, N-methyl β-alanine, norleucine, norvaline, N-propylglycine, O-methyl-homoserine, ornithine , pipecolic acid, pyroglutamic acid, sarcosine, selenocysteine, selenohomocysteine, selenomethionine, selenoethionine, taurine, t-leucine, α,β-diaminopropionic acid, α,γ-diaminobutyric acid, α-aminoisobutyric acid, α-amino-n-butyric acid, α-amino-n-heptanoic acid, α-hydroxy-γ-aminobutyric acid, β-alanine, β-aminoisobutyric acid, β-amino-n-butyric acid, γ-aminobutyric acid, δ-aminolevulinic acid, 1-aminocyclopropane-1-carboxylic acid, azetidine-2-carboxylic acid, cycloleucine, or unnatural amino acids such as pseudoproline. In some embodiments, Y is PABA. In other embodiments, Y is alloisoleucine. In further embodiments, Y is allothreonine. In yet other embodiments, Y is carboxylglutamic acid. In still further embodiments, Y is cystathionine. In other embodiments, Y is D-alanine. In further embodiments, Y is dehydroalanine. In still further embodiments, Y is D-glutamate. In still further embodiments, Y is diaminopimelic acid. In still further embodiments, Y is jenkolic acid. In other embodiments, Y is glycine betaine homocysteine. In still further embodiments, Y is homonorleucine. In still further embodiments, Y is homoserine. In still further embodiments, Y is hydroxyglycine. In other embodiments, Y is hydroxyproline. In still further embodiments, Y is hypusine. In still further embodiments, Y is isoserine. In still further embodiments, Y is isovaline.In another embodiment, Y is lanthionine. In a further embodiment, Y is N-ethylalanine. In yet another embodiment, Y is N-ethylglycine. In even further embodiments, Y is N-ethyl β-alanine. In another embodiment, Y is N-isopropylglycine. In a further embodiment, Y is N-methylalanine. In yet another embodiment, Y is N-methyl β-alanine. In even further embodiments, Y is norleucine. In another embodiment, Y is norvaline. In a further embodiment, Y is N-propylglycine. In yet another embodiment, Y is O-methyl-homoserine. In even further embodiments, Y is ornithine. In another embodiment, Y is pipecolic acid. In a further embodiment, Y is pyroglutamic acid. In yet another embodiment, Y is sarcosine. In even further embodiments, Y is selenocysteine. In another embodiment, Y is selenohomocysteine. In a further embodiment, Y is selenomethionine. In yet another embodiment, Y is selenoethionine. In an even further embodiment, Y is taurine. In another embodiment, Y is t-leucine. In an even further embodiment, Y is α,β-diaminopropionic acid. In an even further embodiment, Y is α,γ-diaminobutyric acid. In an even further embodiment, Y is α-aminoisobutyric acid. In an even further embodiment, Y is α-amino-n-butyric acid. In an even further embodiment, Y is α-amino-n-heptanoic acid. In an even further embodiment, Y is α-hydroxy-γ-aminobutyric acid. In an even further embodiment, Y is β-alanine. In an even further embodiment, Y is β-aminoisobutyric acid. In an even further embodiment, Y is β-amino-n-butyric acid. In an even further embodiment, Y is γ-aminobutyric acid. In an even further embodiment, Y is δ-aminolevulinic acid. In an even further embodiment, Y is 1-aminocyclopropane-1-carboxylic acid. In an even further embodiment, Y is azetidine-2-carboxylic acid. In yet another embodiment, Y is cycloleucine.In an even further embodiment, Y is pseudoproline.

[0094] A deprotecting agent, as described above, is then added for a time and under conditions effective to produce a compound of formula XI-A, XI-B, or XI-C, respectively. Z is a leaving group, which can be selected by one of skill in the art. In some embodiments, Z is halo, sulfonate, N-hydroxybenzotriazolyl, 1-hydroxy-7-azabenzotriazolyl, or one of the following: [ka] In another embodiment, Z is a halo, such as chloro, fluoro, or bromo. In a further embodiment, Z is a sulfonate, such as p-toluenesulfonate (OTs), methanesulfonate (OMs), or trifluoromethanesulfonate (OTf). In yet another embodiment, Z is N-hydroxybenzotriazolyl. In an even further embodiment, Z is 1-hydroxy-7-azabenzotriazolyl. In another embodiment, Z is one of the following: [ka] It is.

[0095] The compound of formula XI-A is then reacted with a compound of formula XII-A: [ka] may be contacted with a weak acid for a time and under conditions sufficient to produce In some embodiments, a compound of formula XII-A is prepared.

[0096] The weak acid utilized in the transformation may be selected by one of skill in the art. In some embodiments, the weak acid is trifluoroacetic acid or acetic acid. In other embodiments, the weak acid is trifluoroacetic acid. In further embodiments, the weak acid is acetic acid. In some embodiments, the weak acid has a pH of about 1 to about 7. In some embodiments, the weak acid has a pH of about 2 to about 6. In some embodiments, the weak acid has a pH of about 2 to about 5. In some embodiments, the weak acid has a pH of about 3 to about 4. In one embodiment, the weak acid has a pH of about 7. In one embodiment, the weak acid has a pH of about 6. In one embodiment, the weak acid has a pH of about 5. In one embodiment, the weak acid has a pH of about 4. In one embodiment, the weak acid has a pH of about 3. In one embodiment, the weak acid has a pH of about 2. In one embodiment, the weak acid has a pH of about 1.

[0097] In some embodiments, the weak acid has a concentration of about 50 to about 200 mM. In even further embodiments, the weak acid has a concentration of about 50, about 75, about 100, about 125, about 150, about 175, or about 200 mM. In other embodiments, the weak acid has a concentration of about 50 to about 175, about 50 to about 150, about 50 to about 125, about 50 to about 100, about 50 to about 75, about 75 to about 200, about 75 to about 175, about 75 to about 150, about 75 to about 125, about 75 to about 100, about 100 to about 200, about 100 to about 175, about 100 to about 150, about 100 to about 125, about 125 to about 200, about 125 to about 175, about 125 to about 150, about 150 to about 200, about 150 to about 175, or about 175 to about 200 mM. In a further embodiment, the weak acid is about 50 to about 200 mM acetic acid.

[0098] The weak acid may include a solvent, examples of which include, but are not limited to, methanol, ethanol, isopropanol, and / or butyl alcohol, or the like, and / or polar aprotic solvents such as dimethylformamide, N-methylpyrrolidinone, and the like.

[0099] The time required to prepare the spirocyclic compounds described herein, including compounds of formula XII-A, ranges from about 1 minute to about 24 hours, preferably from about 2 to about 12 hours. In some embodiments, the time is about 1 minute, about 5 minutes, about 15 minutes, about 30 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, or about 24 hours. In other embodiments, the time is from about 1 minute to about 18 hours, from about 1 minute to about 12 hours, from about 1 minute to about 8 hours, from about 1 minute to about 4 hours, from about 1 minute to about 1 hour, from about 1 minute to about 30 minutes, from about 30 minutes to about 24 hours, from about 30 minutes to about 18 hours, from about 30 minutes to about 12 hours, from about 30 minutes to about 8 hours, from about 30 minutes to about 4 hours, from about 30 minutes to about 1 hour, from about 1 hour to about 24 hours, from about 1 hour to about about 18 hours, about 1 hour to about 12 hours, about 1 hour to about 8 hours, about 1 hour to about 4 hours, about 4 hours to about 24 hours, about 4 hours to about 18 hours, about 4 hours to about 12 hours, about 4 hours to about 8 hours, about 8 hours to about 24 hours, about 8 hours to about 18 hours, about 8 hours to about 12 hours, about 12 hours to about 24 hours, about 12 hours to about 18 hours, or about 18 hours to about 24 hours.

[0100] The temperature utilized in preparing the compound of formula XII-A ranges from room temperature to elevated temperatures. The term "room temperature" as used herein refers to a temperature of about 30 to about 35° C. In some embodiments, the temperature is about 30 to about 60° C. In other embodiments, the temperature is about 30, about 35, about 40, about 45, about 50, about 55, or about 60° C. In further embodiments, the temperature is about 30 to 55, about 30 to about 50, about 30 to about 45, about 30 to about 40, about 30 to about 35, about 35 to about 60, about 35 to about 55, about 35 to about 50, about 35 to about 45, about 35 to about 40, about 40 to about 60, about 40 to about 55, about 40 to about 50, about 40 to about 45, about 45 to about 60, about 45 to about 55, about 45 to about 50, about 50 to about 60, about 50 to about 55, or about 55 to about 60°C.

[0101] The present invention relates to a compound of formula XV: [ka] Further provided is a process for preparing

[0102] In the structure of formula XV, R 1 , R 2 , R 4 and x are defined herein. w is an integer from 1 to 20. In some embodiments, w is an integer 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. In other embodiments, w is an integer from 1 to 18, 1 to 16, 1 to 14, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 4, 1 to 2, 2 to 20, 2 to 18, 2 to 16, 2 to 14, 2 to 12, 2 to 10, 2 to 8, 2 to 6, 2 to 4, 4 to 20, 4 to 18, 4 to 16, 4 to 14, 4 to 12, 4 to 10, 4 to 8, 4 to 6, 6 to 20, 6 to 18, 6 to 1 In the structure of the compound of formula XV, y is an integer of 0 or 1. In certain embodiments, y is an integer of 0. In other embodiments, y is an integer of 1.

[0103] The process for preparing a compound of formula XV comprises reacting a compound of formula II with a compound of formula VI: [ka] {where x, z, R 1 , R 2 , R 5 , and AA is defined herein}.

[0104] The process is carried out using an activating agent, as defined herein, and a base, as defined herein, to provide an intermediate, which is then reacted with a compound of formula VII: [ka] with a deprotecting agent, as defined herein, for a time and under conditions effective to produce

[0105] The compound of formula VII is then contacted with the compound of formula II (w-1) times, where w is defined herein. The term "w-1" times, as used herein, means contacting a compound of formula VII with a compound of formula II to provide a first intermediate. The first intermediate is then contacted with a compound of formula II to provide a second intermediate, and so on. In some embodiments, the compound of formula VII is contacted with a compound of formula II 1 to 19 times. In some embodiments, the compound of formula VII is contacted with a compound of formula II 1, 2, 3, 4, 5, 6, 7, 8 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 times. In another embodiment, a compound of formula VII is contacted with a compound of formula II, where w is an integer from 1 to 18, 1 to 16, 1 to 14, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 4, 1 to 2, 2 to 20, 2 to 18, 2 to 16, 2 to 14, 2 to 12, 2 to 10, 2 to 8, 2 to 6, 2 to 4, 4 to 20, 4 to 18, 4 to 16, 4 to 14, 4 to 12, 4 to 10, 4 to 8, 4 ~6, 6~20, 6~18, 6~16, 6~14, 6~12, 6~10, 6~8, 8~20, 8~18, 8~16, 8~14, 8~12, 8~10, 10~20, 10~18, 10~16, 10~14, 10~12, 12~20, 12~18, 12~16, 12~14, 14~20, 14~18, 14~16, 16~20, 16~18, or 18~20 times.

[0106] The present invention also relates to a compound of formula I: [ka] wherein x, z, and R 4 also provides a process for preparing {as defined herein.

[0107] The process comprises reacting a compound of formula III: [ka] wherein M is a transition metal having a +2 oxidation state as defined herein. The process is carried out for a time and under conditions effective to produce a compound of formula I, using a chelating agent as defined herein.

[0108] The present invention relates to a compound of formula III: [ka] wherein X, Z, and R 4 The present invention further provides a process for preparing {as defined herein. The process includes contacting a compound of formula IV with 9-fluorenylmethoxycarbonyl chloride for a time and under conditions effective to produce a compound of formula III.

[0109] The present invention relates to a compound of formula IV: [ka] wherein x, z, and R 4 The present invention further provides a process for preparing {as defined herein.

[0110] The process comprises reacting a compound of formula V, as defined herein: [ka] with a source of metal(II). The process is carried out for a time and under conditions effective to produce a compound of formula IV.

[0111] The present invention also relates to a compound of formula VII: [ka] {where x, y, z, R 4 , R 5 and AA also provides a process for preparing {as defined herein}. The process comprises reacting a compound of formula II: [ka] with a compound of formula VI: [ka] The method includes contacting the The process is carried out by preparing an intermediate using an activating agent, as defined herein, and a base, as defined herein; and contacting the intermediate with a deprotecting agent, as defined herein, for a time and under conditions effective to produce a compound of formula VII.

[0112] compound Advantageously, the processes described herein allow for the preparation of an unlimited number of compounds. In certain embodiments, the present invention provides a compound of formula XIV: [ka] to provide. In this structure, R 1 , R 2 Each instance of X, y, and z is independently defined herein.

[0113] A compound of formula II: [ka] {where, y, R 1 , R 2 , and R 4 is as defined herein} or a salt thereof is also provided.

[0114] The present invention also provides a compound of formula II-A, II-B, or II-C: [ka] {where, R 1 and R 4 {as defined herein}. In some embodiments, the compound is a compound of formula II-A. In other embodiments, the compound is a compound of formula II-B. In further embodiments, the compound is a compound of formula II-C.

[0115] In yet other embodiments, the compound is: [ka] [ka] [ka] It is.

[0116] In still further compounds, the present invention provides the following: [ka] The present invention provides a compound,

[0117] The present invention relates to the following: [ka] The present invention further provides a compound wherein

[0118] The present invention also relates to a method for producing a composition comprising the steps of: [ka] Also provided is a compound wherein:

[0119] Although the present invention has been described in conjunction with its preferred specific embodiments, it should be understood that the foregoing description and the following examples are intended to illustrate the present invention and are not intended to limit the scope of the present invention. It will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the scope of the present invention, and that other aspects, advantages and modifications will be apparent to those skilled in the art to which the present invention pertains. In addition to the embodiments described herein, the present invention contemplates and claims inventions resulting from combinations of the features of the present invention recited herein and of the cited prior art references that complement the features of the present invention. The inventions of each patent, patent application, and publication cited or described in this document are hereby incorporated by reference in their entirety for all purposes. EXAMPLES

[0120] Experimental Examples The present invention will be described in more detail by reference to the following experimental examples. These examples are provided for illustrative purposes only and are not intended to be limiting, unless otherwise specified. Thus, the present invention should in no way be understood to be limited to the following examples, but rather to encompass any and all variations that become evident as a result of the teachings provided herein.

[0121] Without further description, it is believed that one of ordinary skill in the art can, using the preceding description and the following illustrative examples, make and use the present invention and practice the claimed methods. As such, the following examples specifically point out preferred embodiments of the present invention, and are not to be construed as limiting in any way the remainder of the disclosure.

[0122] Example 1: Development of Fmoc-protected bis-amino acids for the automated synthesis of highly functionalized spiroligomers "Molecular structure defines function" - this is the most fundamental paradigm of molecular biology (Gutteridge, A. et al., Trends in Biochemical Sciences 2005, 30 (11), 622-629). It is the goal of polymer chemistry to create ever larger molecules with control over their three-dimensional structure and the various functional groups they present (Lenci, E. et al., Chemical Society Reviews 2020, 49 (11), 3262-3277; Lutz, J.-F et al., Science 2013, 341 (6146), 1238149). Stoddart first introduced the concept of "molecular LEGO", which can be programmed to have any desired shape by repeating ring fusions such as belt[n] arenes and kohnkenes (Hill, DJ et al., Chemical Reviews 2001, 101 (12), 3893-4012). More recently, the Bode group has demonstrated the iterative assembly of polycyclic saturated heterocyclic compounds from monomeric building blocks (Saito, F et al., Journal of the American Chemical Society 2019, 141 (13), 5544-5554).

[0123] Spiroligomers are fused ring spiro ladder structures composed of cyclic stereochemically pure bis-amino acid building blocks linked together through diketopiperazine (DKP) rings (Schafmeister, CE et al., Accounts of Chemical Research 2008, 41 (10), 1387-1398). The formation of spirocyclic DKPs enhances the rigidity of the backbone by eliminating single bond rotations in the backbone. Meanwhile, the positions and orientations of the various functional groups on the backbone are determined by the sequence and stereochemistry of the building blocks. Various applications of spiroligomers have been developed, including as catalysts for organic reactions (Parker, MFL et al., Journal of the American Chemical Society 2014, 136 (10), 3817-3827), templates for supramolecular metal-binding complexes (Northrup, JD et al., Journal of Organic Chemistry 2021, 86 (6), 4867-4876), inhibitors of protein-protein interactions (Brown, Z. Z et al., Plos One 2012, 7 (10)), and carbohydrate-binding molecules (Chepyshev, SV et al., Rocky Mountain Regional Meeting, Fort Collins, CO, United States 2020).

[0124] The synthesis of functionalized spiroligomers is challenging because the monomer synthesis is laborious even at the ~600 mmol scale. Previously, the original carboxybenzyl (Cbz), tert-butoxycarbonyl (Boc), and p-nitrobenzyloxycarbonyl (pNZ) protecting groups were used as chain-extending removable protecting groups for proline amines in solid-phase synthesis. These protecting groups were either difficult to remove, such as Cbz, or they limited the choice of resin-linking group to those that led to side reactions during deprotection and resulted in low yields (Cheong, JE, Tetrahedron Letters 2016, 57 (44), 4882-4884; Pfeiffer, CT et al, Tetrahedron Letters 2018, 59 (30), 2884-2888).

[0125] As an excellent temporary protecting group in peptide synthesis, the fluorenylmethyloxycarbonyl (Fmoc) group allows the use of excellent high-yield cleavable resin linkers such as the chlorotrityl linker (Behrendt, R. et al., Journal of Peptide Science 2016, 22 (1), 4-27; Ieronymaki, M. et al., Biopolymers 2015, 104 (5), 506-514; Stathopoulos, P. et al., Journal of Peptide Science 2006, 12 (3), 227-232). In the work described herein, the Fmoc group was effectively incorporated into spiroligomer synthesis. The approach combines the scale-up synthesis of bis-amino acid intermediates on the tens of kilograms scale with an effective one-pot synthetic methodology for replacing the Cbz group with the Fmoc group. Four unique spiroligomers are synthesized using Fmoc / tBu solid-phase synthesis and their three-dimensional structures are confirmed by nuclear magnetic resonance (NMR) spectroscopy.

[0126] To significantly reduce the labor costs of bis-amino acid synthesis and take advantage of economies of scale, a large-scale synthesis was aimed at developing key ketone intermediates 2a and 2b. Compared to the previous synthesis (Cheong, JE, Tetrahedron Letters 2016, 57 (44), 4882-4884), the currently utilized route increased the intermediate synthesis scale by three orders of magnitude (Figure 2). The scaled-up synthesis eliminated the use of the highly toxic Jones reagent (Caron, S. et al., Chemical Reviews 2006, 106(7) at 2943-2989) and replaced it with a (2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO)-mediated trichloroisocyanuric acid (TCCA) oxidation, avoiding downstream issues with impurities encountered at the small scale.

[0127] The scaled-up synthesis also eliminated the explosive isobutylene gas (Wright, SW et al., Tetrahedron Letters 1997, 38 (42), 7345-7348), which was problematic at pilot plant scale, so the tert-butylating agent tert-butyl 2,2,2 trichloroacetimidate (TBTA) was utilized instead. In less than two months, tens of kilograms of stereochemically pure S- and R-enantiomers were produced at a cost of less than $4 per gram in a more environmentally friendly manner than conventional synthesis.

[0128] According to a previously developed protocol (Cheong, JE, Tetrahedron Letters 2016, 57 (44), 4882-4884), the Bucherer-Bergs reaction converted 2a and 2b to a mixture of diastereomeric hydantoins with approximately a 5:1 ratio of (2S,4S) and (2S,4R) stereoisomers for 2a, respectively. A large-scale flash chromatography system using a methylene chloride / isopropanol gradient was utilized to separate the crude hydantoin diastereomers, ~200 g per charge. A solvent recycling system was used to regenerate methylene chloride. Each isolated, stereochemically pure hydantoin product was then hydrolyzed to provide the four optically pure bis-amino acids (3a-d) as previously reported (Cheong, JE, Tetrahedron Letters 2016, 57 (44), 4882-4884). A detailed multi-kilogram scale synthesis is described below.

[0129] Since the Fmoc group is sensitive to base, Fmoc protection of building blocks needs to be performed after hydrolysis of the hydantoin and removal of the Cbz group. Inspired by the selective ω-amino protection of lysine (Malkar, N. B et al., Letters in Peptide Science 2000, 7 (5), 263-267), an effective synthesis was developed to introduce the Fmoc group at the proline nitrogen. It relied on the formation of a dimeric Cu(II) complex containing two carboxylic acids and two α-amino groups, which temporarily blocked the primary amino group. The carboxylic acid at C2 was protected by a tert-butyl group, which prevented it from complexing with copper together with the amine at C1. Thus, the free primary amine and carboxylic acid at C4 were protected from Cu. 2+The proline nitrogen in the building block was then protected by an Fmoc protecting reagent, such as Fmoc-Cl or Fmoc-OSu. The copper complex was then released by a strong chelating agent at the end of the exchange step. This strategy was applied to four bis-amino acid stereoisomers (3a–d; Figure 3).

[0130] Cbz deprotection was carried out by hydrogenolysis using Pd / C in aqueous Na2CO3. The product was used in the subsequent Cu(II) complexation step without purification. 1 / 2 equivalent of CuCl2 was added directly to the suspension to form a complex with the free amine and carboxylic acid at the C4 position of the bis-amino acid, resulting in a dark blue solution that was mixed with Pd / C powder. Under Schotten-Baumann reaction conditions, a slight excess of Fmoc-Cl in EtOAc was added dropwise to the aqueous slurry. This biphasic mixture was stirred vigorously for 2 h, after which the Pd / C was filtered off using a short Celite plug. The upper EtOAc layer turned blue with a much higher intensity than the lower aqueous phase, suggesting the formation of an Fmoc-protected Cu complex (present in the organic phase due to its reduced polarity). Impurities in the aqueous phase were removed by a separatory funnel. The combined organic phase was stirred with aqueous ethylenediaminetetraacetic acid (EDTA) to obtain Cu. 2+ has been removed.

[0131] After 12 h, the blue color had completely transferred from the organic to the aqueous phase, indicating that Cu 2+ Once the Fmoc-protected bis-amino acid building blocks 4a-d were released from the copper complexes, they were precipitated from the two-phase system to form a white crystalline powder, as shown in Figure 25. The pure mono-protected solid products 4a-d were then isolated by vacuum filtration, further leaving behind a small amount of unreacted starting material and impurities in the yellowish EtOAc phase, and the Cu-EDTA by-product in the blue aqueous phase. This four-step procedure did not involve chromatography.

[0132] Prior to solid-phase assembly, reductive alkylation was used to incorporate various functional groups into the building blocks by treating them with the corresponding aldehydes and the mild reducing agent NaBH3CN. This was an advantageous feature of spiroligomer synthesis, when functional groups selected from a large set of aldehydes were introduced into stereochemically pure building blocks to form a fixed amount of each monomer for the solid-phase synthesis of spiroligomers. Each Fmoc bis-amino acid diastereomer was alkylated with various functional groups to give the functionalized building blocks 5a-d (Figure 4).

[0133] The four side chains were chosen to represent a wide range of functionality, including alkyl, fused aromatic, heterocyclic, and aryl halogen groups. For functionalization of Fmoc building blocks and Pfp ester activation, previous methods used for pNZ, Boc, and Cbz building blocks were found to be adaptable to the new Fmoc building blocks (Pfeiffer, CT et al, Tetrahedron Letters 2018, 59 (30), 2884-2888; Brown, Z. Z et al., Biopolymers 2011, 96 (5), 578-585).

[0134] By adding DCM with methanol in a 1:1 ratio, the (2S,4R)- and (2R,4S)-diastereomers are dissolved to improve the efficiency of the reductive alkylation. Slurrying the crude solid onto Celite with the aid of copious amounts of methanol and normal phase chromatography with a DCM gradient of 0-20% methanol proved effective for the purification of the functionalized bis-amino acids with yields ranging from 61% to 90%.

[0135] Consistent with previous findings, the functionalized secondary amine of the quaternary center of each building block is so strongly sterically hindered that it cannot be coupled to activated esters even in large ratios (Brown, ZZ et al., Journal of the American Chemical Society 2008, 130 (44), 14382-14383), which means that it can be used as a monomer in solid-phase synthesis without protecting the secondary amine on 5a-d.

[0136] Dimerization of the previous building blocks was observed using a conventional in-situ activation coupling strategy (Brown, ZZ et al., Journal of the American Chemical Society 2008, 130 (44), 14382-14383). Dimerization was minimized using pentafluorophenol (Pfp-OH) to preactivate the monomers, and gave bench-stable building blocks 6a-d, as shown in Figure 4 (Pfeiffer, CT et al, Tetrahedron Letters 2018, 59 (30), 2884-2888). In the coupling step, the Pfp esters of the building blocks were added neat to the amines on the solid support to avoid the formation of symmetric dimers. Each monomer was purified by normal phase column chromatography in 0-50% hexane / EtOAc and stored at -20 °C until required.

[0137] Highly functionalized spiroligomers were assembled by solid-phase synthesis using the four stereoisomers of building blocks 6a-d on a semi-automated microwave peptide synthesizer. As shown in Figure 6, L-proline was first loaded onto 2-chlorotrityl chloride resin using N,N-diisopropylethylamine (DIPEA) followed by deprotection of the Fmoc group to generate 7. The second residue, Pfp ester 6a, was coupled using a similar HOAt / DIPEA protocol developed for the pNZ building block (Pfeiffer, CT et al, Tetrahedron Letters 2018, 59 (30), 2884-2888).

[0138] After optimization, it was found that 2 equivalents of building block were sufficient to complete the coupling in the presence of 4 equivalents of HOAt and 8 equivalents of DIPEA at 50 °C for 1 h. To balance the acidity of the excess of HOAt, an excess of base was used, which prematurely cleaved the highly sensitive chlorotrityl linker. Following coupling of building block 6b to the resin, Fmoc was removed to give 9. Following coupling of the next bis-amino acid 6c, removal of the Fmoc group was removed to give 10. This process was repeated for the introduction of the last building block 6d, and compound 11 was formed. The sequence was capped with Fmoc-Dab(Boc)-OH to give 12, which was released from the solid phase by exposure to TFA. The DKP of tetrameric T1 was probably formed completely by an acid-catalyzed condensation reaction after heating the TFA solution containing the cleaved product 12 at 40 °C overnight. No significant by-products were observed by crude HPLC (Figure 26). The final yield of T1 was 30% after preparative HPLC, based on maximum loading of the resin.

[0139] To demonstrate the generality of this synthetic approach, three other tetramers, T2-T4, were synthesized by varying the positions of the building blocks (Figure 5). The compositions of T1-T4 were confirmed by high-resolution mass spectrometry (QTOF MS) (Figures 27-30). Two-dimensional nuclear magnetic resonance experiments: double quantum filtering correlation spectroscopy (DQF-COSY) in DMSO-d6, heteronuclear single quantum correlation spectroscopy (HSQC), heteronuclear multiple bond correlation spectroscopy (HMBC), and heteronuclear multiple quantum coherence (HMQC) were performed and analyzed using the software package SPARKY. 1 H and 13 The C resonances were used to assign the expected connectivity with cross peaks from correlations between adjacent building blocks in band-selected HMBC spectra. Rotating frame Overhauser effect spectroscopy (ROESY) correlations were used to identify the relative stereochemistry of the C2 and C4 hydrogens of each pyrrolidine and to confirm their geometry relative to that of the C1 carbon on each building block.

[0140] Furthermore, all side chains had hydrogens that showed ROESY correlations with backbone hydrogens, which were consistent with side chains having preferred orientations. For example, as shown in FIG. 6, one of the β-protons, FHB2, had a strong ROESY correlation with GHa1 (red solid box) under the plane of the second bis-amino acid proline ring, which was one of the methylene protons in the side group of the adjacent building block. Meanwhile, no correlation was found between GHa1-FHB1, GHa2-FHB1, or GHa2-FHB2 (expected blue dashed circle, FIG. 6). This indicated that the rotation of the side chains was restricted by interactions with the rigid backbone, which was consistent with the predictions of a well-defined three-dimensional structure. The energy-minimized structure of T1 is also shown in FIG. 5.

[0141] In summary, the research demonstrated the scale-up of the synthesis of bis-amino acid intermediates to tens of kilograms scale. 2+The complexation strategy was successfully used to selectively incorporate the Fmoc group in a chromatography-free, four-step, one-pot process with excellent yields and purity. Using these monomers, four spiroligomers were synthesized on solid support with four different building blocks containing unique functional groups and stereochemistry. This work laid the foundation for the reliable and automated assembly of highly functionalized spiroligomer libraries.

[0142] Overall, this study demonstrated the efficient multi-kilogram-scale synthesis of bis-amino acid precursors along with the temporary Cu 2+ Fluorenylmethoxycarbonyl (Fmoc) protection of functionalized bis-amino acid building blocks using a complexation strategy is described. This enabled the synthesis of stereochemically and functionally diverse spiroligomers utilizing solid-phase Fmoc / tBu chemistry to facilitate application development. Four tetramers were assembled on a semi-automated microwave peptide synthesizer. Secondary structures were also determined using two-dimensional nuclear magnetic resonance spectroscopy.

[0143] The materials and methods utilized in the experimental examples are described herein. General method Reactions were carried out in standard, oven-dried glassware equipped with PTFE-coated magnetic stir bars. Stainless steel syringes were used to transfer air- and moisture-sensitive liquids. Concentrations reported refer to solution volumes at room temperature. Evaporation in vacuo and concentration were performed using a rotary evaporator.

[0144] material Reagents were purchased from commercial suppliers of reagent grade and used without further purification unless otherwise noted. Compounds S2a, S3a, 2a, S2b, S3b, and 2b were synthesized (Levins, CG et al., Journal of the American Chemical Society 2003, 125 (16), 4702-4703).1 The H NMR spectrum was very clean and compared well with published NMR spectra. Compounds 3a-d were synthesized according to a previously reported procedure (Cheong, JE et al., Tetrahedron Letters 2016, 57 (44), 4882-4884).

[0145] Device Low-resolution high-performance liquid chromatography-mass spectrometry (LR-HPLC-MS) analysis was performed on an Agilent 1290 liquid chromatograph system equipped with a Supelco Ascentis® Express C8 column (2.7 μm packing, 2.1×50 mm) using an acetonitrile-water gradient solvent system containing 0.1% formic acid at a flow rate of 1.0 mL / min. The system was connected to an Agilent 6120 single quadrupole mass spectrometer utilizing electrospray ionization.

[0146] High-resolution HPLC-MS was performed on an Agilent Infinity II series LCMS system equipped with an Agilent Poroshell 120 EC-C18 column (1.9 μm packing, 2.1×50 mm) using an acetonitrile-water gradient containing 0.1% formic acid at a flow rate of 1.0 mL / min. The system was coupled to a 6000 series quadrupole time-of-flight (QTOF) mass spectrometer.

[0147] Preparative HPLC was performed on an Agilent Infinity II series LC / MSD system equipped with a Phenomenex Aeris Peptide XB-C18 column (5 μm packing, 21.2 × 150 mm) using a water-acetonitrile gradient solvent system containing 0.1% trifluoroacetic acid at a flow rate of 25 mL / min with a single quadrupole mass spectrometer utilizing electrospray ionization.

[0148] 1 H and 13C one-dimensional nuclear magnetic resonance (NMR) experiments were performed on a Bruker Avance 500 MHz instrument at 25° C. Chemical shifts were reported relative to residual solvent peaks or tetramethylsilane. Data are expressed as follows: chemical shift, integral, multiplicity (br=broad, s=singlet, d=doublet, t=triplet, q=quintet, qn=sexlet, sp=septet, m=multiplet), coupling constants in Hertz (Hz). Some compounds exist at room temperature as a mixture of two slowly interconverting (on the NMR time scale) rotamers due to tertiary amides. Rotamer peaks are indicated in the NMR data.

[0149] Two-dimensional NMR experiments were performed on a Bruker Avance NEO 600 MHz spectrometer equipped with a cryoprobe at the Spectroscopy Support Facility (SSF) of Fox Chase Cancer Center. Normal phase purification was performed on an ISCOR (Teledyne, Inc.) automated flash chromatography system using prepacked RediSep® Rf silica gel columns of various sizes (60 Å porosity, 230×400 mesh particle size) purchased from Sorbent Technologies.

[0150] The hydantoin diastereomers were separated on a Biotage® Isolera LS automated flash purification system using a Biotage® SNAP KP-SIL 1500 g column (55 Å porosity, 50 μm particle size). Reverse-phase purification was performed on an ISCO (Teledyne, Inc.) automated flash chromatography system using prepacked RediSep® Rf C18 reverse-phase columns of various sizes (60 Å porosity, 230×400 mesh particle size) purchased from Sorbent Technologies, using a water-acetonitrile gradient solvent system containing 0.1% formic acid. Solid-phase synthesis of the spiroligomers was carried out on a DiscoverBio semi-automated microwave-assisted peptide synthesizer from CEM Corporation equipped with a 25 mL reaction vessel.

[0151] software 1D NMR spectra were processed using Bruker Topspin. 2D NMR spectra were analyzed using SPARKY (Goddard, TD; Kneller, DG Sparky 3.190; University of California: San Francisco, CA, 2015). (Lee, W. et al., Bioinformatics 2014, 31 (8), 1325-1327). Molecular images were generated using UCSF Chimera 1.14 (build42094; Pettersen, EF et al., Journal of Computational Chemistry 2004, 25 (13), 1605-1612). The in-house software package CANDO (github.com / cando-developers / cando.git) was used to perform energy minimization of the three-dimensional structure of T1 using the Generalized Amber Force Field (GAFF) energy function (Wang, J. et al., Journal of Computational Chemistry 2004, 25 (9), 1157-1174).

[0152] Abbreviation DIPEA = diisopropylethylamine, DIC = N,N'-diisopropylcarbodiimide, ESI = electrospray ionization, HOAt = 1-hydroxy-7-azabenzotriazole, TFA = trifluoroacetic acid, THF = tetrahydrofuran, DMF = dimethylformamide, DMSO = dimethylsulfoxide, MTBE = methyl tert-butyl ether, PyAOP = ((7-azabenzotriazol-1-yloxy)tripyrrolidino-phosphonium hexafluorophosphate), Fmoc = fluorenylmethoxycarbonyl (fluoren phenylmethoxycarbony), Boc = tert-butyloxycarbonyl, Cbz-Cl = benzyl chloroformate, Pfp = pentafluorophenyl, EDTA = ethylenediaminetetraacetic acid, Dab = 2,4-diaminobutyric acid, pro = proline, NMR = nuclear magnetic resonance, TEMPO = (2,2,6,6-tetramethylpiperidin-1-yl)oxyl or oxydanyl, TLC = thin layer chromatography, DKP = diketopiperazine, HPLC = high performance liquid chromatography, HR = high resolution, LC = liquid chromatography, LR = low resolution, MS = mass spectrometry.

[0153] Experimental procedures and characterization data Synthesis and characterization of building blocks Multikilogram-scale synthesis of 2a [ka]

[0154] Synthesis of S2a [ka] (2S,4R)-1-((benzyloxy)carbonyl)-4-hydroxypyrrolidine-2-carboxylic acid (S2a) - Purification yield: 27.7 kg, 91% Five reactions were carried out in parallel. To a solution of compound 1a (3.00 kg, 22.8 mol) and NaHCO3 (4.80 kg, 57.2 mol) in H2O (18.0 L) was added CbzCl (3.71 kg, 21.7 mol) in THF (3 L), and the mixture was stirred at 20 °C for 1 h. TLC (petroleum ether / ethyl acetate = 3 / 1, Rf = 0.00) showed that CbzCl was completely consumed, and TLC (dichloromethane / methanol = 5 / 1, Rf = 0.20) detected the product. The five reactions were combined for workup. The reaction mixture was cooled to 0 °C, and HCl (12 M, 600.0 mL, pH = 2) was added dropwise. The mixture was then extracted with EtOAc (8.00 L x 2). The organic portion was washed with brine (5.00 L) and dried over Na2SO4. The mixture was concentrated in vacuo to give compound S2a as a yellow oil. 1 H NMR (400 MHz, DMSO, observing rotamers) δ 7.29 - 7.38 (m, 5H), 5.04 - 5.12 (m, 2H), 4.21 - 4.30 (m, 2H), 3.36 - 3.47 (m, 2H), 2.12 - 2.20 (m, 1H), 1.88 - 1.98 (m, 1H).

[0155] Synthesis of S3a [ka] (S)-1-((benzyloxy)carbonyl)-4-oxopyrrolidine-2-carboxylic acid (S3a) - Purification yield: 15.0 kg, 92% Five reactions were carried out in parallel. To a solution of compound S2a (5.00 kg, 18.8 mol) in EtOAc (25.0 L) was added TEMPO (148.0 g, 942 mmol) and trichloroisocyanuric acid (3.07 kg, 13.1 mol) at 0 °C. The mixture was stirred at 0 °C for 3 h. TLC (dichloromethane / methanol = 10 / 1, Rf = 0.4) showed that the starting material was completely consumed. The five reactions were combined for workup. The reaction mixture was filtered to remove insoluble material. The mixture was quenched by adding Na2S2O3 (20.0 L) at 0-10 °C. The combined organic layer was washed with brine (10.0 L x 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The crude product was triturated with petroleum ether / MTBE=1 / 1 (10.0 L) at 25° C. for 1 h, filtered, and the solvent was evaporated to give compound S3a as a white solid. 1 H NMR (400 MHz, CDCl3, observing rotamers) δ 9.05 - 9.09 (m, 1H), 7.26 - 7.37 (m, 5H), 5.16 -5.27 (m, 2H), 4.86 - 4.92 (m, 1H), 3.89 - 4.02 (m, 2H), 2.94 - 3.06 (m, 1H), 2.67 - 2.77 (m, 1H).

[0156] Synthesis of 2a [ka] 1-Benzyl-2-(tert-butyl)(S)-4-oxopyrrolidine-1,2-dicarboxylate (2a) - Purification yield: 10.0 kg, 69% Four reactions were carried out in parallel. To a solution of compound S3a (3.00 kg, 11.4 mol) in THF (12.0 L) and cyclohexane (12.0 L) was added tert-butyl 2,2,2-trichloroacetimidate (4.98 kg, 22.7 mol) at 12° C. Diethyloxonio(trifluoro)boranuide (161.0 g, 1.14 mol) was added dropwise to the mixture at 0° C. The mixture was stirred at 12° C. for 2 h. TLC (dichloromethane / methanol=10 / 1, Rf=0.8) showed that the starting material was completely consumed. The four reactions were combined for workup. The mixture was quenched with NaHCO3 (4.00 L), adjusted to pH=8, then washed with brine (5.00 L) and dried over Na2SO4. The organic layer was concentrated to give a residue. The residue was filtered to remove insoluble materials, and the solution was concentrated in vacuo to give compound 2a as a brown oil. 1 H NMR (400 MHz, CDCl3, observing rotamers) δ 7.32 - 7.38 (m, 5H), 5.12 - 5.26 (m, 2H), 4.68 - 4.76 (m, 1H), 3.88 - 4.02 (m, 2H), 2.90 - 3.00 (m, 1H), 2.52 - 2.58 (m, 1H), 1.41 (s, 9H, rotamerism). HRMS (ESI-TOF) m / z:C 17 H 21 NO5Na [M+Na] + Calculated value 342.1312; measured value 342.1311.

[0157] Synthesis of 2b on a multi-kilogram scale [ka]

[0158] Synthesis of S2b [ka] (2R,4R)-1-((benzyloxy)carbonyl)-4-hydroxypyrrolidine-2-carboxylic acid (S2b) - Purification yield: 15.0 kg, 82% Three reactions were carried out in parallel. To a solution of compound 1b (3.00 kg, 22.8 mol) and NaHCO3 (4.80 kg, 57.2 mol) in H2O (18.0 L) was added CbzCl (3.71 kg, 21.7 mol) in THF (3.00 L), and the mixture was stirred at 20 °C for 3 h. TLC (petroleum ether / ethyl acetate = 3 / 1, Rf = 0.00) showed that CbzCl was completely consumed, and TLC (dichloromethane / methanol = 5 / 1, Rf = 0.30) detected the product. Three reactions were combined for workup. The reaction mixture was cooled to 0 °C, and HCl (12 M, 600.0 mL, until pH = 2) was added dropwise. The mixture was then extracted with EtOAc (8.00 L x 2). The organic portion was washed with brine (5.00 L) and dried over Na2SO4. The mixture was concentrated in vacuo to give compound S2b as a white solid. 1 H NMR (400 MHz, DMSO-d6, observing rotamers) δ 7.30 - 7.37 (m, 5H), 5.02 - 5.08 (m, 2H), 4.19 - 4.28 (m, 2H), 3.55 - 3.59 (m, 1H), 3.19 - 3.23 (m, 1H), 2.32 - 2.40(m, 1H), 1.89 - 1.93 (m, 1H).

[0159] Synthesis of S3b [ka] (R)-1-((benzyloxy)carbonyl)-4-oxopyrrolidine-2-carboxylic acid (S3b) - Purification yield: 12.0 kg, 94% Three reactions were carried out in parallel. To a solution of compound S2b (4.30 kg, 16.21 mol) in DCM (28.0 L) was added TEMPO (127.5 g, 810.5 mmol) and trichloroisocyanuric acid (2.64 kg, 11.4 mol) at 0 °C. The mixture was stirred at 0 °C for 3 h. TLC (dichloromethane / methanol = 10 / 1, Rf = 0.40) showed that the starting material was completely consumed. The three reactions were combined for workup. The reaction mixture was filtered to remove insoluble material. The mixture was quenched by adding Na2S2O3 (20.0 L) at 0-10 °C. The combined organic layer was washed with brine (10.0 L x 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The crude product was triturated with petroleum ether / MTBE=1 / 1 (10.0 L) at 25° C. for 1 h, filtered, and the solvent was evaporated to give compound S3b as a white solid. 1 H NMR (400 MHz, CDCl3, observing rotamers) δ 9.15 (br, 1H), 7.32 - 7.35 (m, 5H), 5.15 - 5.28 (m, 2H), 4.84 - 4.90 (m, 1H), 3.88 - 4.00 (m, 2H), 2.92 - 2.99(m, 1H), 2.70 - 2.74 (m, 1H).

[0160] Synthesis of 2b [ka] 1-Benzyl-2-(tert-butyl) (R)-4-oxopyrrolidine-1,2-dicarboxylate (2b) - Purification yield: 13.0 kg, 89% Four reactions were carried out in parallel. To a solution of compound S3b (3.00 kg, 11.4 mol) in THF (12.0 L) and cyclohexane (12.0 L) was added tert-butyl 2,2,2-trichloroacetimidate (4.98 kg, 22.7 mol) at 12° C. Diethyloxonio(trifluoro)boranuide (161.0 g, 1.14 mol) was added dropwise to the mixture at 0° C. The mixture was stirred at 12° C. for 2 hours. TLC (dichloromethane / methanol=10 / 1, Rf=1.00) showed that the starting material was completely consumed. TLC (petroleum ether / ethyl acetate=3 / 1, Rf=0.5) showed that the product was detected. The four reactions were combined for workup. The mixture was quenched with NaHCO3 (4.00 L), adjusted to pH=8, then washed with brine (5.00 L) and dried over Na2SO4. The organic layer was concentrated to give a residue. The residue was filtered to remove insoluble material, and the solution was concentrated in vacuo to give compound 2b as a brown oil. By WuXi Apptec 1 H NMR (400 MHz, CDCl3, observing rotamers) δ 7.32 - 7.36 (m, 5H), 5.13 - 5.24 (m, 2H), 4.69 - 4.75 (m, 1H), 3.90 - 4.02 (m, 2H), 2.90 - 2.98 (m, 1H), 2.52 - 2.57 (m, 1H), 1.40 (s, 9H, rotamerism). HRMS (ESI-TOF) m / z:C 17 H 21 NO5Na [M+Na] + Calculated value 342.1312; measured value 342.1312.

[0161] Synthesis of compounds 4a-d [ka] (3S,5S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-amino-5-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid (4a) - Purification yield: 12.2 g, 71% Compounds 3a-d (38 mmol) were dissolved in Na2CO3 (300 mL, 0.2 mol / L). To this solution, Pd / C (10% wt) was added and a balloon filled with H2 was attached via a three-way valve. The atmosphere was removed by evacuation and back-filling with H2 several times. After stirring at room temperature overnight, the reaction reached completion (monitored by LCMS). To the stirred solution, CuCl2 (20 mmol) was added and allowed to react for 30 min. Fmoc-Cl (40 mmol) dissolved in EtOAc (100 mL) was added dropwise to the reaction mixture and stirred vigorously for another 2 h. Pd / C was removed by vacuum filtration using Celite, and the organic layer was collected, washed with saturated NaCl solution (250 mL x 2), and transferred to a beaker. The solution was then mixed with EDTA sodium salt solution (250 mL, 0.2 M) and stirred vigorously overnight. The products 4a-d were isolated under vacuum filtration as white solids. This four-step reaction sequence was carried out without any chromatography. 1 H NMR (500 MHz, DMSO) δ 8.78 (s, 2H), 7.91 (t, J = 7.4 Hz, 2H), 7.64 (d, J = 7.5 Hz, 2H, rotamerism), 7.43 (t, J = 7.3 Hz, 2H), 7.34 - 7.31 (m, 2H), 4.42 - 4.15 (m, 4H, rotamerism), 4.04 (d, J = 11.5 Hz, 1H, rotamerism), 3.58 (d, J = 11.5 Hz, 1H, rotamerism), 2.88 (dd, J = 13.4, 8.5 Hz,1H, rotamerism), 2.21 (dd, J = 13.4, 8.8 Hz, 1H, rotamer), 1.40 (s, 9H). 13C NMR (125 MHz DMSO, rotamers observed) δ 172.1, 171.7, 170.8, 170.7, 154.1, 154.0, 144.2, 144.1, 144.0, 143.7, 141.2, 128.3, 128.2, 127.6, 125.9, 125.7, 125.6, 120.6, 81.9, 81.5, 67.7, 67.3, 62.0, 61.0, 60.2, 59.9, 55.1, 54.3, 47.1, 46.9, 38.9, 37.9, 28.1, 28.00;HRMS (ESI-TOF) m / z:C 25 H 29 N2O6 [M+H] + Calculated value 453.2020; measured value 453.2016.

[0162] [ka] (3R,5S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-amino-5-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid (4b) - Purification yield: 13.0 g, 74% 1 H NMR (500 MHz, DMSO) δ 8.74 (s, 2H), 7.91 (d, J = 7.5 Hz, 2H), 7.72 - 7.66 (m, 2H), 7.44 (t, J = 7.4 Hz, 2H), 7.36 - 7.31 (m, 2H), 4.61, 4.41 (m, 1H, rotamerism), 4.40 - 4.04 (m, 2H, rotamerism), 3.99 (m, 1H, rotamerism), 3.85, 3.76 (d, J = 12.2 Hz, 1H, rotamerism), 2.77, 2.67 (dd, J = 14.1, 9.0 Hz,1H, rotamerism), 2.50 (dd, J = 14.3, 7.8 Hz, 1H, rotamerism), 1.40 (d, J = 5.3 Hz, 9H); 13C NMR (125 MHz, DMSO, rotamers observed) δ 170.9, 170.3, 169.9, 163.6, 154.0, 153.8, 144.1, 144.0, 143.7, 141.2, 141.1, 128.3, 128.2, 127.7, 127.6, 125.9, 125.8, 125.7, 120.7, 120.6, 82.2, 81.8, 68.0, 67.7, 63.9, 63.0, 58.9, 58.4, 55.0, 54.4, 47.0, 46.9, 38.1, 28.0;HRMS (ESI-TOF) m / z:C 25 H 29 N2O6 [M+H] + Calculated value: 453.2020, measured value: 453.2028.

[0163] [ka] (3R,5R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-amino-5-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid (4c) - Purification yield: 10.9 g, 64% 1 H NMR (500 MHz, DMSO) δ 8.78 (s, 2H), 7.91 (t, J = 7.4 Hz, 2H), 7.64 (d, J = 7.5 Hz, 2H, rotamerism), 7.43 (t, J = 7.3 Hz, 2H), 7.34 - 7.31 (m, 2H), 4.42 - 4.15 (m, 4H, rotamerism), 4.04 (d, J = 11.4 Hz, 1H, rotamerism), 3.58 (d, J = 11.5 Hz, 1H, rotamerism), 2.88, 2.84 (dd, J = 13.3, 8.5 Hz,1H, rotamerism), 2.24, 2.21 (dd, J = 13.3, 8.8 Hz, 1H, rotamer), 1.41 (s, 9H); 13C NMR (125 MHz, DMSO, rotamers observed) δ 172.1, 171.7, 170.9, 170.8, 154.1, 154.0, 144.2, 144.1, 144.0, 143.7, 141.1, 128.3, 128.2, 127.6, 125.9, 125.7, 125.6, 121.9, 120.6, 120.5, 81.9, 81.5, 67.7, 67.3, 62.0, 61.0, 60.2, 59.9, 55.1, 54.3, 47.1, 46.9, 38.9, 37.9, 28.1, 28.0;HRMS (ESI-TOF) m / z:C 25 H 29 N2O6 [M+H] + Calculated value: 453.2020, measured value: 453.2017.

[0164] [ka] (3R,5S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-3-amino-5-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid (4d) - Purification yield: 14.7g, 85% 1 H NMR (500 MHz, DMSO) δ 8.75 (s, 2H), 7.91 (d, J = 7.6 Hz, 2H), 7.72 - 7.66 (m, 2H), 7.44 (t, J = 7.5 Hz, 2H), 7.36 - 7.31 (m, 2H), 4.61, 4.41 (m, 1H, rotamerism), 4.40 - 4.04 (m, 2H, rotamerism), 3.99 (m, 1H, rotamerism), 3.85, 3.78 (d, J = 12.2 Hz, 1H, rotamerism), 2.77, 2.67 (dd, J = 14.1, 7.7 Hz,1H, rotamerism), 2.53, 2.45 (dd, J = 14.3, 7.6 Hz, 1H, rotamerism), 1.40 (d, J = 5.3 Hz, 9H); 13C NMR (125 MHz, DMSO, observing rotamers) δ 170.8, 170.3, 169.9, 163.6, 154.0, 153.8, 144.1, 143.7, 141.2, 141.1, 128.4, 128.2, 127.7, 127.6, 125.9, 125.8, 125.7, 120.7, 82.2, 81.8, 68.0, 67.7, 63.8, 62.9, 58.8, 58.4, 54.9, 54.6, 47.0, 38.1, 28.0 (ESI-TOF) m / z:C 25 H 29 N2O6 [M+H] + Calculated value: 453.2020, actual value: 453.2029.

[0165] Synthesis of compounds 5a-d [ka]

[0166] Synthesis of functionalized bis-amino acids 5a [ka] (3S,5S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-5-(tert-butoxycarbonyl)-3-((naphthalen-2-ylmethyl)amino)pyrrolidine-3-carboxylic acid (5a) - Purification yield: 1.82 g, 61% 2-Naphthaldehyde (7.5 mmol) was added to a 100 mL round bottom flask containing compound 4a (5 mmol) dissolved in 25 mL MeOH. The solution was stirred for 30 min, at which point NaBH3CN (7.5 mmol) was added and stirred overnight. The reaction progress was monitored by LCMS, and if incomplete, additional aldehyde and reducing agent were added. Upon completion of the reductive alkylation, the solution was transferred to a round bottom flask containing 10 g of Celite. The solvent was removed under reduced pressure, and the dry powder was then transferred to a packed cartridge for flash chromatography. Normal phase separation was performed with a mobile phase of DCM / 20% MeOH in DCM using a 0-100% gradient. Fractions containing pure product were collected and the solvent was removed under reduced pressure to give the functionalized bis-amino acid 5a. Purity was confirmed by LCMS. 1 H NMR (500 MHz, DMSO) δ 7.90 - 7.83 (m, 6H), 7.68 - 7.63 (m, 2H), 7.53 - 7.38 (m, 5H), 7.34 - 7.28 (m, 2H), 4.39, 4.32 (m, 1H, rotamerism), 4.23 (m, 2H, rotamerism), 4.15 (m, 1H, rotamerism), 4.00, 3.90 (m, 1H, rotamerism), 3.88 (m, 1H), 3.82 (m, 1H), 3.52, 3.46 (d, J = 10.8 Hz, 1H, rotamerism), 2.76 - 2.68 (m, 1H, rotamer), 2.15, 2.07 (dd, J = 12.8, 6.0 Hz, 1H, rotamer), 1.36 (s, 9H); 13C NMR (125 MHz, DMSO) δ 174.4, 171.2, 170.8, 154.3, 144.3, 144.1, 143.9, 141.1, 137.2, 133.3, 132.7, 128.2, 128.0, 127.6, 127.4, 127.0, 126.5, 126.1, 125.8, 125.7, 125.6, 120.6, 81.3, 81.0, 68.2, 67.5, 67.3, 59.7, 59.3, 54.9, 54.4, 49.0, 47.2, 46.9, 39.1, 38.0, 28.1;HRMS (ESI-TOF) m / z:C 36 H 37 N2O6 [M+H] + Calculated value 593.2624, measured value 593.2638.

[0167] Synthesis of functionalized bis-amino acids 5b [ka] (3R,5S)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-5-(tert-butoxycarbonyl)-3-(isopentylamino)pyrrolidine-3-carboxylic acid (5b) - Purification yield: 2.34 g, 90% Isovaleraldehyde (7.5 mmol) was added to a 100 mL round bottom flask containing compound 4b (5 mmol) suspended in 25 mL MeOH / DCM. The solution was stirred for 30 min at which point NaBH3CN (7.5 mmol) was added and stirred overnight. The reaction progress was monitored by LCMS and, if incomplete, additional aldehyde and reducing agent were added. Upon completion of the reductive alkylation, the solution was transferred to a round bottom flask containing 10 g of Celite. The solvent was removed under reduced pressure and the dry powder was then transferred to a packed cartridge for flash chromatography. Normal phase separation was performed with a mobile phase of 20% MeOH in DCM / DCM using a 0-100% gradient. Fractions containing pure product were collected and the solvent was removed under reduced pressure to give the functionalized bis-amino acid 5b. Purity was confirmed by LCMS. 11H NMR (500 MHz, DMSO) δ 7.91 - 7.89 (m, 2H), 7.68 - 7.64 (m, 2H), 7.44 - 7.41 (m, 2H), 7.35 - 7.30 (m, 2H), 4.55, 4.49 (dd, J = 9.1, 5.6 Hz, 1H, rotamer), 4.47 - 4.45 (m, 1H), 4.27, 4.23 (m, 1H, rotamer), 4.15 (m, 1H, rotamer), 4.05 (t, J = 13.6 Hz, 1H), 3.84 (d, J = 12.7 Hz, 1H, rotamer), 3.07 - 2.83 (m, 3H), 2.63, 2.55 (dd, J = 14.8, 5.4 Hz, 1H, rotamer), 1.67 - 1.61 (m, 1H), 1.53 - 1.49 (m, 2H), 1.40 (s, 9H, rotamer), 0.89 (d, J = 6.6 Hz, 6H); 13 13C NMR (125 MHz, DMSO) δ 170.2, 169.9, 169.4, 169.3, 153.8, 153.7, 144.1, 143.7, 141.2, 128.3, 127.6, 125.7, 125.5, 120.6, 119.2, 116.9, 114.6, 112.3, 82.3, 81.9, 69.0, 68.1, 67.9, 58.9, 58.5, 53.5, 52.9, 47.0, 46.9, 43.0, 36.6, 35.3, 27.9, 25.7, 22.3; HRMS (ESI-TOF) m / z: C 30 H 39 [M + H] of C + Calculated value 523.2803, measured value 523.2797.

[0168] Synthesis of Functionalized Bis-Amino Acid 5c

Chemical Structure

[0169] Synthesis of functionalized bis-amino acid 5d [ka] (3S,5R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-5-(tert-butoxycarbonyl)-3-((3,4-dichlorobenzyl)amino)pyrrolidine-3-carboxylic acid (5d) - Purification yield: 2.67g, 87% 2-Naphthaldehyde (7.5 mmol) was added to a 100 mL round bottom flask containing compound 4d (5 mmol) suspended in 25 mL MeOH. The solution was stirred for 30 min at which point NaBH3CN (7.5 mmol) was added and allowed to proceed overnight. The reaction progress was monitored by LCMS and, if incomplete, additional aldehyde and reducing agent were added. Upon completion of the reductive alkylation, the solution was transferred to a round bottom flask containing 10 g of Celite. The solvent was removed under reduced pressure and the dry powder was then transferred to a packed cartridge for flash chromatography. Normal phase separation was performed with a mobile phase of DCM / 20% MeOH in DCM using a 0-100% gradient. Fractions containing pure product were collected and the solvent was removed under reduced pressure to give the functionalized bis-amino acid 5d. Purity was confirmed by LCMS. 1H NMR (500 MHz, DMSO) δ 7.91 - 7.86 (m, 2H), 7.68 (d, J = 7.5 Hz, 1H, rotamerism), 7.59 - 7.49 (m, 3H), 7.43 - 7.22 (m, 5H), 4.36 - 4.27 (m, 1H, rotamerism), 4.22 - 4.11, 4.04 (m, 3H, rotamerism), 3.73 - 3.45 (m, 4H), 2.34, 2.10 (m,1H, rotamerism), 2.21 (dd, J = 12.8, 8.5 Hz, 1H, rotamerism), 1.38 (s, 9H, rotamerism); 13 C NMR (125 MHz, DMSO) δ 174.8, 174.7, 172.1, 171.6, 154.5, 154.4, 144.3, 144.1, 143.7, 141.1, 131.2, 131.1, 130.6, 130.5, 130.1, 130.0, 129.4, 129.3, 128.5, 128.3, 128.1, 127.6, 127.5, 125.8, 125.6, 125.5, 125.4, 120.7, 120.6, 81.3, 80.9, 69.4, 68.6, 67.5, 67.0, 59.9, 59.4, 55.2, 54.2, 47.2, 47.1, 46.9, 38.9, 28.1, 28.0;HRMS (ESI-TOF) m / z:C 32 H 33 Cl2N2O6 [M+H] + Calculated value 611.1710, measured value 611.1714.

[0170] Synthesis of compounds 6a-d [ka]

[0171] Synthesis of functionalized pentafluorophenyl esters 6a [ka] 1-((9H-fluoren-9-yl)methyl) 2-(tert-butyl) 4-(perfluorophenyl) (2S,4S)-4-((naphthalen-2-ylmethyl)amino)pyrrolidine-1,2,4-tricarboxylate (6a) - Purified yield: 1.07 g, 71% Compound 5a (2 mmol) and pentafluorophenol (10 mmol) were weighed and dissolved in 10 mL of DCM. DIC (4 mmol) was added and the reaction was allowed to stir overnight. The reaction was confirmed to be complete by LCMS and then transferred to a 50 mL round bottom flask containing Celite (5 g). Removal of the solvent under reduced pressure at room temperature gave a white powder. A cartridge was loaded with dry Celite and flash chromatography was performed using hexane / EtOAc with a 5-100% EtOAc gradient. The fractions containing the pure product were collected under reduced pressure to give 6a as a yellowish solid. 1 H NMR (500 MHz, CDCl3, observing rotamers) δ 7.83 - 7.74 (m, 6H), 7.64 - 7.57 (m, 2H), 7.51 - 7.46 (m, 3H), 7.42 - 7.36 (m, 2H), 7.32 - 7.25 (m, 2H), 4.56 - 4.46 (m, 2H), 4.38 - 4.21 (m, 3H), 4.00 - 3.84 (m, 3H), 2.94 (m, 1H), 2.50 (dt, J = 12.9, 4.7 Hz, 1H, rotamerism), 1.43 (s, 9H, rotamerism); 13C NMR (125 MHz, CDCl3, observing rotamers) δ 170.2, 170.1, 169.7, 169.6, 154.4, 144.2, 144.0, 143.7, 143.5, 141.3, 141.2, 136.0, 135.9, 133.4, 132.9, 128.3, 127.8, 127.7, 127.1, 127.0, 126.9, 126.6, 126.5, 126.2, 125.9, 125.3, 125.2, 125.1, 125.0, 120.0, 82.2, 82.1, HRMS (ESI-TOF) m / z:C 42 H 36 F5N2O6 [M+H] + Calculated value: 759.2423, measured value: 759.2488.

[0172] Synthesis of functionalized pentafluorophenyl esters 6b [ka] 1-((9H-fluoren-9-yl)methyl) 2-(tert-butyl) 4-(perfluorophenyl) (2S,4R)-4-(isopentylamino)pyrrolidine-1,2,4-tricarboxylate (6b) - Purified yield: 1.01 g, 73% Compound 5b (2 mmol) and pentafluorophenol (10 mmol) were weighed and dissolved in 10 mL of DCM. DIC (4 mmol) was added and the reaction was allowed to stir overnight. The reaction was confirmed to be complete by LCMS and then transferred to a 50 mL round bottom flask containing Celite (5 g). Removal of the solvent under reduced pressure at room temperature gave a white powder. A cartridge was loaded with dry Celite and flash chromatographed using hexane / EtOAc with a 5-100% EtOAc gradient. The fractions containing the pure product were collected under reduced pressure to give 6b (50.7%) as a white solid.1 H NMR (500 MHz, CDCl3, observing rotamers) δ 7.78 - 7.76 (m, 2H), 7.69 - 7.59 (m, 2H), 7.43 - 7.39 (m, 2H), 7.34 - 7.30 (m, 2H), 4.55 - 4.20 (m, 4H), 4.05 (t, J = 13.6 Hz, 1H), 4.02, 3.84 (m, 1H, rotamerism), 2.63 - 2.43 (m, 4H), 1.46 (s, 9H, rotamerism);1.41 (q, J = 7.2 Hz, 2H), 1.22 (d, J = 6.4 Hz, 1H), 0.92 - 0.89 (m, 6H); 13 C NMR (125 MHz, CDCl3, rotamers observed) δ 171.0, 170.7, 168.8, 168.7, 154.7, 144.2, 144.0, 143.8, 143.5, 141.4, 141.3, 127.8, 127.7, 127.2, 127.1, 125.5, 125.2, 125.1, 120.0, 82.2, 82.0, 68.2, 68.1, 67.7, 67.4, 59.2, 58.9, 54.6, 54.1, 47.2, 47.1, 42.7, 42.7, 39.4, 39.4, 38.9, 37.8, 28.0, 26.0, 25.9, 22.6, 22.5;HRMS (ESI-TOF) m / z:C 36 H 38 F5N2O6 [M+H] + Calculated value 689.2645, measured value 689.2649.

[0173] Synthesis of functionalized pentafluorophenyl ester 6c [ka] 1-((9H-fluoren-9-yl)methyl) 2-(tert-butyl) 4-(perfluorophenyl) (2R,4R)-4-((pyridin-4-ylmethyl)amino)pyrrolidine-1,2,4-tricarboxylate (6c) - Purified yield: 0.752 g, 53% Compound 5c (2 mmol) and pentafluorophenol (10 mmol) were weighed and dissolved in 10 mL of DCM. DIC (4 mmol) was added and the reaction was allowed to stir overnight. The reaction was confirmed to be complete by LCMS and then transferred to a 50 mL round bottom flask containing Celite (5 g). Removal of the solvent under reduced pressure at room temperature gave a white powder. A cartridge was loaded with dry Celite and flash chromatography was performed using hexanes / EtOAc with a 5-100% EtOAc gradient. The fractions containing the pure product were collected under reduced pressure to give 6c as a white solid. 1 H NMR (500 MHz, CDCl3, observing rotamers) δ 8.58 - 8.45 (m, 2H), 7.78 - 7.75 (m, 2H), 7.64 - 7.56 (m, 2H), 7.42 - 7.37 (m, 4H), 7.32 - 7.25 (m, 2H), 4.55 - 4.47 (m, 2H), 4.41 - 4.33 (m, 1H), 4.29 - 4.10 (m, 2H), 3.89 - 3.74 (m, 3H), 2.96 - 2.86 (m, 1H, rotamerism), 2.47 (td, J = 13.8, 3.9 Hz, 1H), 1.44 (d, 9H, rotamer); 13 C NMR (125 MHz, CDCl3, rotamers observed) δ 175.6, 175.5, 171.1, 171.0, 170.2, 169.2, 169.1, 154.4, 149.5, 149.4, 148.5, 148.4, 147.6, 144.1, 143.9, 143.6, 143.4, 141.3, 127.8, 127.2, 127.1, 127.0, 125.2, 125.1, 125.0, 123.6, 123.5, 120.0, 82.5, 82.4, 68.7, 67.7, 58.8, 58.6, 54.4, 54.0, 48.1, 47.3, 47.1, 39.5, 38.2, 28.1;HRMS (ESI-TOF) m / z:C 37 H 33 F5N3O6 [M+H]+ Calculated value: 710.2284, measured value: 710.2289.

[0174] Synthesis of functionalized pentafluorophenyl ester 6d [ka] 1-((9H-fluoren-9-yl)methyl) 2-(tert-butyl) 4-(perfluorophenyl) (2R,4S)-4-((3,4-dichlorobenzyl)amino)pyrrolidine-1,2,4-tricarboxylate (6d) - Purified yield: 1.07 g, 71% Compound 5d (2 mmol) and pentafluorophenol (10 mmol) were weighed and dissolved in 10 mL of DCM. DIC (4 mmol) was added and the reaction was allowed to stir overnight. The reaction was confirmed to be complete by LCMS and then transferred to a 50 mL round bottom flask containing Celite (5 g). Removal of the solvent under reduced pressure at room temperature gave a white powder. A cartridge was loaded with dry Celite and flash chromatographed using hexane / EtOAc with a 5-100% EtOAc gradient. The fractions containing the pure product were collected under reduced pressure to give 6d as a white solid. 1 H NMR (500 MHz, CDCl3, observing rotamers) δ 7.78 - 7.51 (m, 4H), 7.45 - 7.14 (m, 7H), 4.58 - 4.45 (m, 2H, rotamers), 4.39 - 4.30 (m, 1H, rotamers), 4.22 (t, J = 7.1 Hz, 1H), 4.10 - 4.02 (m, 1H), 3.97 - 3.64 (m,3H), 2.63 - 2.46 (m, 2H), 1.46 (s, 9H, rotamerism); 13C NMR (125 MHz, CDCl3, rotamers observed) δ 170.7, 170.5, 168.4, 168.3, 154.8, 154.6, 144.1, 143.9, 143.7, 143.4, 141.3, 141.3, 139.2, 139.1, 132.7, 131.6, 130.5, 129.9, 129.8, 127.8, 127.2, 127.1, 125.4, 125.1, 125.0, 124.9, 120.1, 120.0, 119.9, 82.5, 82.3, 68.2, 67.5, 59.1, 58.9, 54.3, 53.6, 47.7, 47.6, 47.2, 47.1, 39.3, 38.2, 28.0;HRMS (ESI-TOF) m / z:C 38 H 32 Cl2F5N2O6 [M+H] + Calculated value: 777.1481, measured value: 777.1556.

[0175] Solid-phase synthesis of spiroligomers General methods for solid phase synthesis General method of resin filling Solid-phase synthesis of the spiroligomer was carried out by microwave peptide synthesizer. Cl-TCP(Cl) protide resin (100 mg, 0.05 mmol, 0.5 mmol / g loading) was weighed into a 25 mL reaction vessel. Fmoc-Pro-OH was weighed into a 2 mL centrifuge tube and dissolved in anhydrous DMF (2 mL) with DIPEA (168 μL, 1 mmol). This solution was then added to the resin in the reaction vessel in one portion and reacted at 50° C. for 30 min with Ar bubbling. After draining the solution, the resin was washed with 2 mL of DMF three times. Unreacted trityl chloride was capped with 2 mL of capping solution containing 1:3:17 methanol / DIPEA / DCM for 10 min at room temperature, and the resin was washed with DMF three times.

[0176] General procedure for Fmoc deprotection All deprotections were achieved with 20% piperidine in DMF (2 mL) (1×4 min, 60° C.). After each Fmoc deprotection, the resin was washed with DMF (3×1 mL).

[0177] A general method for coupling building blocks A solution of the building block (0.1 mmol, 2 eq.) in 1 mL of anhydrous DMF was added to the resin in a reaction vessel. Then, a solution of HOAt (27.2 mg, 0.2 mmol) and DIPEA (69.7 μL, 0.4 mmol) in 1 mL of anhydrous DMF was added. The reaction mixture was heated at 50° C. for 1 h. Completion of the reaction was confirmed by test cleavage using LCMS after washing with DMF.

[0178] General procedure for coupling of Fmoc-Dab(Boc)-OH Fmoc-Dab(Boc)-OH (63.9 mg, 0.15 mmol, 3 eq.) in 1 mL of DMF was transferred to a reactor with a mixture of PyAOP (78.2 mg, 0.15 mmol), DIPEA (52.3 mL, 0.3 mmol), and DMF (2 mL), and they were reacted for 5 min at 50° C. Completion of the reaction was confirmed by test cleavage using LCMS after washing with DMF.

[0179] A general method for cleavage and closure of DKPs Final cleavage was achieved by using TFA (2 mL x 2) for 30 min at room temperature. The product was collected in a 50 mL centrifuge tube and transferred to a 20 mL glass vial. It was placed in a heat block with stirring at 40°C overnight.

[0180] Solid-phase synthesis of T1 The first residue Fmoc-Pro-OH (169 mg, 0.5 mmol) was loaded onto resin (100 mg, 0.05 mmol, 0.5 mmol / g loading) using the general procedure for resin loading and deprotected using the general procedure for Fmoc deprotection. The general procedure for building block coupling and Fmoc deprotection was utilized with 6a (75.8 mg, 0.1 mmol, 2 equiv.), followed by 6b (68.8 mg, 0.1 mmol), 6c (71 mg, 0.1 mmol), and 6d (77.3 mg, 0.1 mmol). Next, Fmoc-Dab(Boc)-OH (63.9 mg, 0.15 mmol) was coupled and deprotected as described previously. Finally, cleavage of T1 and DKP closure were performed according to the general procedure. After confirming the completion of DKP closure by LCMS, the product was precipitated with cold diethyl ether and purified by preparative HPLC (25%-95% gradient of MeCN in water with 0.1% TFA). The purified fractions containing the product were collected and then lyophilized to give spiroligomer T1 as a white powder. T1-purified yield, 18 mg, 30%; HPLC purity, 98%; retention time, 3.122 min. HRMS (ESI-TOF) m / z: C 62 H 67 Cl2N 12 O 10 [M+H] + Calculated value: 1209.44, actual value: 1209.44.

[0181] Solid-phase synthesis of T2 The first residue Fmoc-Pro-OH (169 mg, 0.5 mmol) was loaded onto resin (100 mg, 0.05 mmol, 0.5 mmol / g loading) using the general procedure for resin loading and deprotected using the general procedure for Fmoc deprotection. The general procedure for building block coupling and Fmoc deprotection was utilized with 6d (77.3 mg, 0.1 mmol, 2 equiv.), followed by 6c (71 mg, 0.1 mmol), 6b (68.8 mg, 0.1 mmol), and 6a (75.8 mg, 0.1 mmol). Next, Fmoc-Dab(Boc)-OH (63.9 mg, 0.15 mmol) was coupled and deprotected as described previously. Finally, cleavage of T2 and DKP closure were performed according to the general procedure. After confirming the completion of DKP closure by LCMS, the product was precipitated with cold diethyl ether and purified by preparative HPLC (25%-95% gradient of MeCN in water with 0.1% TFA). The purified fractions containing the product were collected and then lyophilized to give spiroligomer T2 as a white powder. T2-purified yield, 16 mg, 27%; HPLC purity, 96%; retention time, 3.297 min. HRMS (ESI-TOF) m / z: C 62 H 66 Cl2N 12 O 10 [M+H] + Calculated value: 1209.44, actual value: 1209.45.

[0182] Solid-phase synthesis of T3 The first residue Fmoc-Pro-OH (169 mg, 0.5 mmol) was loaded onto resin (100 mg, 0.05 mmol, 0.5 mmol / g loading) using the general procedure for resin loading and deprotected using the general procedure for Fmoc deprotection. The general procedure for building block coupling and Fmoc deprotection was utilized with 6b (68.8 mg, 0.1 mmol, 2 equiv.), followed by 6d (77.3 mg, 0.1 mmol), 6a (75.8 mg, 0.1 mmol), and 6c (71 mg, 0.1 mmol). Next, Fmoc-Dab(Boc)-OH (63.9 mg, 0.15 mmol) was coupled and deprotected as described previously. Finally, T3 cleavage and DKP closure were performed according to the general procedure. After confirming the completion of DKP closure by LCMS, the product was precipitated with cold diethyl ether and purified by preparative HPLC (25%-95% gradient of MeCN in water with 0.1% TFA). The purified fractions containing the product were collected and then lyophilized to give spiroligomer T3 as a white powder. T3-purified yield, 15 mg, 24%; HPLC purity, 99%; retention time, 3.515 min. HRMS (ESI-TOF) m / z: C 62 H 67 Cl2N 12 O 10 [M+H] + Calculated value: 1209.44, actual value: 1209.45.

[0183] Solid-phase synthesis of T4 The first residue Fmoc-Pro-OH (169 mg, 0.5 mmol) was loaded onto resin (100 mg, 0.05 mmol, 0.5 mmol / g loading) using the general procedure for resin loading and deprotected using the general procedure for Fmoc deprotection. The general procedure for building block coupling and Fmoc deprotection was utilized with 6c (71 mg, 0.1 mmol, 2 equiv.), followed by 6d (77.3 mg, 0.1 mmol), 6a (75.8 mg, 0.1 mmol), and 6b (68.8 mg, 0.1 mmol). Next, Fmoc-Dab(Boc)-OH (63.9 mg, 0.15 mmol) was coupled and deprotected as described previously. Finally, T4 cleavage and DKP closure were performed according to the general procedure. After confirming the completion of DKP closure by LCMS, the product was precipitated with cold diethyl ether and purified by preparative HPLC (25%-95% gradient of MeCN in water with 0.1% TFA). The purified fractions containing the product were collected and then lyophilized to give spiroligomer T4 as a white powder. T4-purified yield, 19 mg, 32%; HPLC purity, 99%; retention time, 3.333 min. HRMS (ESI-TOF) m / z: C 62 H 67 Cl2N 12 O 10 [M+H] + Calculated 1209.44, found 1209.45. (2S,4R)-1-((benzyloxy)carbonyl)-4-hydroxypyrrolidine-2-carboxylic acid (S2a).

[0184] The disclosures of any and all patents, patent applications, and publications cited herein are hereby incorporated by reference in their entirety. Although the present invention has been disclosed with respect to specific embodiments, it is apparent that other embodiments and variations of the present invention may be devised by those skilled in the art without departing from the true spirit and scope of the present invention. It is intended that the appended claims be construed to include equivalent variations of such embodiments.

Claims

1. A compound of formula I: 【Chemistry 1】 {During the ceremony, R 1 is an amino acid, a β-amino acid, a sugar, an optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 selected from the group consisting of alkynyl, optionally substituted aryl, and any combination thereof; R 2 is absent or is selected from H, amino acids, β-amino acids, sugars, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 selected from the group consisting of alkynyl, optionally substituted aryl, and any combination thereof; or R 1 and R 2 are concatenated to form C 3-8 Forming a cycloalkyl; X is a leaving group; R 4 is a protecting group; x is an integer 1 or 2; and z is an integer 1 or 2; However, both x and z cannot be 2. , R 1 X, R 1 C(O)H or R 1 C(O)R 2 and, In the presence of a reducing agent; A compound of formula II: 【Chemistry 2】 {During the ceremony, y is an integer 0 or 1; provided that when the compound of formula I is 1 When reacting with X, y is an integer 0 and R 2 is non-existent. contacting the compound for a time and under conditions effective to produce

2. R 1 and R 2 But C 0-6 Alkyl OH, C 0-6 Alkyl SH, C 0-6 Alkyl NH 2 , C 0-6 Alkyl-OC 0-6 Alkyl, C 0-6 Alkyl-SC 0-6 Alkyl, C 0-6 Alkyl C(O)OH, C 0-6 AlkylC(O)(C 1-6 alkyl), C 0-6 AlkylC(O)O(C 1-6 alkyl), C 0-6 Alkyl N 3 , C 0-6 AlkylC(O)NH 2 , C 0-6 Alkyl-C(O)N(C 1-6 alkyl)OH, optionally substituted C 3-7 Cycloalkyl, optionally substituted C 5-7 Cycloalkenyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, -NHC(NH 2 )(=N(C 0-6 alkyl), -NHC(NH 2 )(=S(C 0-6 alkyl), nucleobase, or amino acid; wherein any carbon atom of said alkyl, alkenyl, or alkynyl may be replaced by a heteroatom, i.e., O, S, SO, SO 2 , or NR 7 optionally replaced by R 7 But H, C 1-4 Alkyl, C 3-4 Alkenyl, C 3-4 Alkynyl, or C 1-4 bridged alkyl, and any combination thereof; wherein a bridge is formed between a nitrogen and a carbon atom of said heteroatom-containing chain to form a ring; The ring is optionally Ar 1 fused to; and Ar 1 is optionally replaced by C 3-6 2. The process of claim 1, wherein the aryl is selected from the group consisting of cycloalkyl, heterocyclyl, aryl, heteroaryl, and any combination thereof.

3. The Ar 1 is the group consisting of phenyl, 1-naphthyl, 2-naphthyl, indenyl, azulenyl, fluorenyl, and anthracyl; or 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, 2-pyrazolinyl, pyrazolidinyl, isoxazolyl, isotriazolyl, 1,2,3-oxadiazolyl, 1,2,3-triazolyl, 1,3,4-thiadiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,3,5-triazinyl, 1,3,5-trithianyl, indolizinyl, indolyl, isoindolyl, 3 3. The process of claim 2, wherein the heteroaromatic group is selected from the group consisting of H-indolyl, indolinyl, benzo[b]furanyl, benzo[b]thiophenyl, 1H-indazolyl, benzimidazolyl, benzthiazolyl, purinyl, 4H-quinolidinyl, quinolinyl, 1,2,3,4-tetrahydroquinolinyl, isoquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 1,8-naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, and any combination thereof.

4. The Ar 1 One or more of H, halo, OH, NO 2 , -SO 3 H, CF 3 , OCF 3 , C 1-6 Alkyl, C 2-6 Alkenyl, C 1-6 Alkoxy, OC 3-4 Alkenyl, -O-benzyl, -O-phenyl, 1,2-methylenedioxy, -NR 5 R 6 , -C(O)OH, -C(O)(C 1-6 alkyl), -C(O)O(C 1-6 alkyl), -C(O)NH(C 1-6 alkyl), -C(O)NH(C 3-5 alkenyl), -C(O)N(C 1-6 Alkyl)(C 1-6 alkyl), -C(O)(C 3-5 alkenyl)(C 3-6 alkenyl), morpholinyl, piperidinyl, -O-Ar 2 , -CH 2 -(CH 2 ) q -Ar 2 , -O-(CH 2 ) q -Ar 2 , -(CH 2 ) q -O-Ar 2 , or -CH=CH-Ar 2 optionally replaced by; R 5 and R 6 However, independently, H, C 1-6 Alkyl, C 3-6 Alkenyl, C 3-6 selected from the group consisting of alkynyl, or benzyl, and any combination thereof; Ar 2 is selected from the group consisting of 4-methoxyphenyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, pyrazyl, quinolyl, 3,5-dimethylisoxazolyl, 2-methylthiazolyl, thiazolyl, 2-thienyl, 3-thienyl or pyrimidinyl, and any combination thereof; and 3. The process of claim 2, wherein q is an integer from 0 to 2.

5. R 1 and R 2 2. The process of claim 1, wherein one or both of are selected from the group consisting of an amino acid, a β-amino acid, a sugar, or any combination thereof.

6. R 4 C 1-6 2. The process of claim 1, wherein the alkyl group is alkyl.

7. R 4 2. The process of claim 1, wherein is selected from the group consisting of t-butyl, 4-{N-[1-(4,4-dimethyl-2,6-dioxocyclohexylidene)-3-methylbutyl]amino}benzyl ester (DMab), and combinations thereof.

8. 2. The process of claim 1, wherein X is selected from the group consisting of halogen and sulfonate.

9. 2. The process of claim 1, wherein X is selected from the group consisting of chloro, fluoro, bromo, p-toluenesulfonate (OTs), methanesulfonate (OMs), and trifluoromethanesulfonate (OTf).

10. The reducing agent is NaBH 3 , NaBH 3 CN, and Na(CH 3 COO) 3 2. The process of claim 1, wherein the compound is selected from the group consisting of BH.

11. The compound of formula I has the structure of formula IA, IB, or IC: 【Transformation 3】 2. The process of claim 1, comprising:

12. The compound of formula II has the structure of formula II-A: 【Chemistry 4】 2. The process of claim 1, comprising:

13. below: a) a compound of formula III: 【Transformation 5】 {wherein M is a transition metal having a +2 oxidation state}; a chelating agent; contacting for a time and under conditions effective to produce a compound of formula I; 10. The process of claim 1, further comprising preparing a compound of formula I by:

14. 14. The process of claim 13, wherein M is selected from the group consisting of vanadium, manganese, iron, cobalt, nickel, copper, and zinc.

15. 14. The process of claim 13, wherein the chelating agent is selected from the group consisting of ethylenediaminetetraacetic acid (EDTA) or a salt thereof, diethylenetriaminepentaacetic acid (DTPA) or a salt thereof, and hydroxyethylethylenediaminetriacetic acid (HEDTA) or a salt thereof.

16. The salt of EDTA is Na 2 EDTA-H 2 O and CaNa 2 16. The process of claim 15, wherein the anionic surfactant is selected from the group consisting of EDTA.

17. The compound of formula III has the structure of formula III-A: 【Transformation 6】 14. The process of claim 13, wherein the compound has the formula:

18. The compound of formula III has the structure of formula III-B, III-C, III-D, III-E, III-F, or III-G: 【Chemistry 7-1】 【Chemistry 7-2】 【Transformation 7-3】 14. The process of claim 13, wherein the compound has the formula:

19. below: a) a compound of formula IV: 【Transformation 8】 with 9-fluorenylmethoxycarbonyl chloride; contacting for a time and under conditions effective to produce a compound of formula IV; 14. The process of claim 13, further comprising preparing the compound of formula III by:

20. The compound of formula IV has the structure of formula IV-A: 【Chemistry 9】 20. The process of claim 19, wherein the compound has the formula:

21. The compound of formula IV has the structure of formula IV-B, IV-C, IV-D, IV-E, IV-F, or IV-G: 【Chemistry 10】 20. The process of claim 19, wherein the compound has the formula:

22. below: a) a compound of formula V: 【Chemistry 11】 with a metal(II) source; contacting for a time and under conditions effective to produce a compound of formula IV; 20. The process of claim 19, further comprising preparing a compound of formula IV by:

23. 23. The process of claim 22, wherein the metal(II) source is selected from the group consisting of a Ni(II) source or a salt thereof and a copper(II) source or a salt thereof.

24. 23. The process of claim 22, wherein the metal source is a copper (II) source or a salt thereof.

25. 25. The process of claim 24, wherein the copper (II) salt is selected from the group consisting of copper chloride and copper sulfate.

26. The compound of formula V has the structure of formula VA, VB, or VC: 【Chemistry 12】 23. The process of claim 22, wherein the compound has the formula:

27. below: a) reacting a compound of formula II with a compound of formula VI-A, VI-B, or VI-C: 【Chemistry 13】 where AA is an optionally substituted amino acid and R 5 is a resin} together with an activating agent and a base to provide an intermediate; and b) reacting the intermediate with a compound of formula VII-A, VII-B, or VII-C: 【Chemistry 14】 with a deprotecting agent for a time and under conditions effective to produce 10. The process of claim 1, further comprising:

28. 28. The process of claim 27, wherein the resin comprises a cleavable group.

29. 29. The process of claim 28, wherein the cleavable group is selected from the group consisting of OH, carboxyl, and polystyrene resin.

30. The activator may be selected from the group consisting of 1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (COMU), 1H-1,2,3-benzotriazol-1-yloxy-tris(pyrrolidino)-phosphonium hexafluorophosphate (PyBOP), 1-hydroxy-7-azabenzotriazole (HOAt), 2-(1H-7-azabenzotriazol-1-yl)-1, 1,3-Tetramethyluronium hexafluorophosphate (HATU), Benzotriazol-1-yl-oxy-tris(dimethylamino)phosphonium hexafluorophosphate (BOP), Chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate, Dicyclohexylcarbodiimide (DCC), Diisopropylcarbodiimide (DIC), Ethyl cyano(hydroxyimino)acetate-O 2 28. The process of claim 27, wherein the hydroxybenzotriazole is selected from the group consisting of 1H-benzotriazole-1-yl-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU), 1H-benzotriazole-1-yl-N,N,N',N'-tetramethyluronium hexafluorophosphate (HCTU), 1H-benzotriazole-1-yl-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), and any combination thereof.

31. 28. The process of claim 27, wherein the deprotecting agent is a base.

32. 32. The process of claim 31, wherein the base is selected from the group consisting of piperidine, 4-methylpiperidine, piperazine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), and morpholine.

33. 28. The process of claim 27, wherein the amino acid is a naturally occurring amino acid.

34. 34. The process of claim 33, wherein the naturally occurring amino acid is selected from the group consisting of arginine (Arg), histidine (His), lysine (Lys), aspartic acid (Asp), glutamic acid (Glu), serine (Ser), threonine (Thr), asparagine (Asn), glutamine (Gln), cysteine ​​(Cys), selenocysteine ​​(Sec), glycine (Gly), proline (Pro), alanine (Ala), valine (Val), isoleucine (Ile), leucine (Leu), methionine (Met), phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp), and pyrrolysine.

35. The amino acid may be 4-aminobenzoic acid (PABA), alloisoleucine, allothreonine, carboxyglutamic acid, cystathionine, D-alanine, dehydroalanine, D-glutamate, diaminopimelic acid, dengkol acid, glycine betaine, homocysteine, homonorleucine, homoserine, hydroxyglycine, hydroxyproline, hypusine, isoserine, isovaline, lanthionine, N-ethylalanine, N-ethylglycine, N-ethyl β-alanine, N-isopropylglycine, N-methylalanine, N-methyl β-alanine, norleucine, norvaline, N-propylglycine, O-methyl-homoserine, 28. The process of claim 27, wherein the amino acid is selected from the group consisting of ornithine, pipecolic acid, pyroglutamic acid, sarcosine, selenocysteine, selenohomocysteine, selenomethionine, selenoethionine, taurine, t-leucine, α,β-diaminopropionic acid, α,γ-diaminobutyric acid, α-aminoisobutyric acid, α-amino-n-butyric acid, α-amino-n-heptanoic acid, α-hydroxy-γ-aminobutyric acid, β-alanine, β-aminoisobutyric acid, β-amino-n-butyric acid, γ-aminobutyric acid, δ-aminolevulinic acid, 1-aminocyclopropane-1-carboxylic acid, azetidine-2-carboxylic acid, cycloleucine, and pseudoproline.

36. The compound of formula VII-A, formula VII-B, or formula VII-C has the following structure of formula VII-A', VII-B', or VII-C': 【Chemistry 15】 28. The process of claim 27, wherein the compound has the formula:

37. The compound of formula VII-A, VII-B, or VII-C can be converted into a compound of formula VIII-A, VIII-B, or VIII-C, respectively, by reacting the compound of formula II and an activating agent followed by a deprotecting agent: 【Chemistry 16】 28. The process of claim 27, further comprising contacting for a time and under conditions effective to produce

38. The compound of formula VIII-A, VIII-B, or VIII-C can be converted to a compound of formula IX-A, IX-B, or IX-C, respectively, by reacting the compound of formula II with an activating agent followed by a deprotecting agent: 【Chemistry 17】 38. The process of claim 37, further comprising contacting for a time and under conditions effective to produce

39. A compound of formula IX-A, IX-B, or IX-C can be converted into a compound of formula XA, XB, or XC, respectively, by reacting a compound of formula II and an activating agent followed by a deprotecting agent. [Chemistry 18] 39. The process of claim 38, further comprising contacting for a time and under conditions effective to produce

40. A compound of formula XA, XB, or XC is reacted with an activator and YZ {During the ceremony, Y is hydrogen or an aminocarbonyl group; and Z is a leaving group. followed by a deprotecting agent to afford a compound of formula XI-A, XI-B, or XI-C, respectively: 【Chemistry 19】 40. The process of claim 39, further comprising contacting for a time and under conditions effective to produce

41. 41. The process of claim 40, wherein Y is selected from the group consisting of H and an amino acid.

42. 41. The process of claim 40, wherein Z is selected from the group consisting of halogen, sulfonate, and triazolyl.

43. Z is chloro, fluoro, bromo, p-toluenesulfonate (OTs), methanesulfonate (OMs), trifluoromethanesulfonate (OTf), N-hydroxybenzotriazolyl, 1-hydroxy-7-azabenzotriazolyl, and the following: 【Chemistry 20】 41. The process of claim 40, wherein the compound is selected from the group consisting of:

44. The compound of formula XI-A is reacted with a weak acid to produce a compound of formula XII-A: 【Chemistry 21】 41. The process of claim 40, further comprising contacting for a sufficient time and under sufficient conditions to produce

45. 45. The process of claim 44, wherein the time period is from about 1 minute to about 24 hours.

46. 45. The process of claim 44, wherein the time is from about 2 to about 12 hours.

47. 45. The process of claim 44, wherein the temperature is from about room temperature to elevated temperature.

48. 48. The process of claim 47, wherein the elevated temperature is at least about 60°C.

49. 45. The process of claim 44, wherein the weak acid is selected from the group consisting of trifluoroacetic acid, acetic acid, and any combination thereof.

50. The contacting step is repeated w times to obtain a compound of formula XV-A, XV-B, or XV-C: 【Chemistry 22】 {wherein w is an integer from 1 to 20} 40. The process of claim 39, comprising obtaining

51. 51. The process of claim 50, wherein w is an integer from 1 to 8.

52. 51. The process of claim 50, wherein w is an integer from 6 to 8.

53. A compound produced using the process of any one of claims 1 to 52.

54. The compound has the structure of Formula XIV: 【Chemistry 23】 54. The compound of claim 53, wherein the compound has the formula:

55. A compound of formula I: 【Chemistry 24】 {During the ceremony, R 4 is a protecting group; x is an integer 1 or 2; and z is an integer 1 or 2; However, both x and z cannot be 2.

1. A process for preparing a compound of formula III: 【Chemistry 25】 where M is a transition metal having a +2 oxidation state. of, with a chelating agent for a time and under conditions effective to produce a compound of formula I.

56. A compound of formula III: 【Chemistry 26】 {During the ceremony, R 4 is a protecting group; x is an integer 1 or 2; and z is an integer 1 or 2; However, both x and z cannot be 2.

1. A process for preparing a compound of formula III comprising contacting a compound of formula IV with 9-fluorenylmethoxycarbonyl chloride for a time and under conditions effective to produce a compound of formula III.

57. A compound of formula IV: 【Chemistry 27】 1. A process for preparing a compound of formula V: 【Chemistry 28】 with a source of metal(II) for a time and under conditions effective to produce a compound of formula IV.

58. A compound of formula VII: 【Chemistry 29】 1. A process for preparing a compound of formula II: 【Transformation 30】 together with an activating agent and a base to form a compound of formula VI: 【Chemistry 31】 {During the ceremony, AA is an optionally substituted amino acid; and R 5 is a resin} to prepare an intermediate; and contacting the intermediate with a deprotecting agent for a time and under conditions effective to produce a compound of formula VII.

59. A compound of formula XV: 【Chemistry 32】 {During the ceremony, R 1 is optionally replaced by C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 selected from the group consisting of alkynyl, optionally substituted aryl, and any combination thereof; R 2 is absent or H, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 selected from the group consisting of alkynyl, optionally substituted aryl, and any combination thereof; or R 1 and R 2 are concatenated, and C 3-8 Forming a cycloalkyl; R 4 is a protecting group; x is an integer 1 or 2; and z is an integer 1 or 2, provided that x and z are not both 2; w is an integer from 1 to 20; and y is an integer 0 or 1. A process for preparing below: a) reacting a compound of formula II below with an activating agent and a base to produce a compound of formula VI below: 【Transformation 33】 {During the ceremony, AA is an optionally substituted amino acid; and R 5 is a resin} to prepare an intermediate; b) reacting the intermediate with a deprotecting agent to form a compound of formula VII: 【Transformation 34】 contacting for a time and under conditions effective to produce c) contacting the compound of formula VII with the compound of formula II (w-1) times; The process comprising:

60. A compound of formula II: 【Chemistry 35】 {During the ceremony, y is an integer 0 or 1; R 1 is optionally replaced by C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 selected from the group consisting of alkynyl, optionally substituted aryl, and any combination thereof; R 2 is absent or H, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 selected from the group consisting of alkynyl, optionally substituted aryl, and any combination thereof; or R 1 and R 2 are concatenated, and C 3-8 Forming a cycloalkyl; R 4 is a protecting group} or a pharmaceutically acceptable salt thereof.

61. R 1 and R 2 But C 0-6 Alkyl OH, C 0-6 Alkyl SH, C 0-6 Alkyl NH 2 , C 0-6 Alkyl-OC 0-6 Alkyl, C 0-6 Alkyl-SC 0-6 Alkyl, C 0-6 Alkyl C(O)OH, C 0-6 AlkylC(O)(C 1-6 alkyl), C 0-6 AlkylC(O)O(C 1-6 alkyl), C 0-6 Alkyl N 3 , C 0-6 AlkylC(O)NH 2 , C 0-6 Alkyl-C(O)N(C 1-6 alkyl)OH, optionally substituted C 3-7 Cycloalkyl, optionally substituted C 5-7 Cycloalkenyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, -NHC(NH 2 )(=N(C 0-6 alkyl), -NHC(NH 2 )(=S(C 0-6 alkyl), nucleobase, or amino acid; wherein any carbon atom of the alkyl, alkenyl, or alkynyl is O, S, SO, SO 2 , and NR 7 optionally substituted with a heteroatom selected from the group consisting of: R 7 But H, C 1-4 Alkyl, C 3-4 Alkenyl, C 3-4 Alkynyl, or C 1-4 and any combination thereof, wherein a bridge is formed between a nitrogen and a carbon atom of said heteroatom-containing chain to form a ring, and wherein said ring is optionally selected from the group consisting of Ar 1 fused to; Ar 1 is optionally replaced by C 3-6 61. The compound of claim 60, wherein the compound is selected from the group consisting of cycloalkyl, heterocyclyl, aryl, heteroaryl, and any combination thereof.

62. The Ar 1 is the group consisting of phenyl, 1-naphthyl, 2-naphthyl, indenyl, azulenyl, fluorenyl, and anthracyl; or 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, 2-pyrazolyl, pyrazolidinyl, isoxazolyl, isotriazolyl, 1,2,3-oxadiazolyl, 1,2,3-triazolyl, 1,3,4-thiadiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,3,5-triazinyl, 1,3,5-trithianyl, indolizinyl, indolyl, isoindolyl, 3 62. The compound of claim 61, wherein the heteroaromatic group is selected from the group consisting of H-indolyl, indolinyl, benzo[b]furanyl, benzo[b]thiophenyl, 1H-indazolyl, benzimidazolyl, benzthiazolyl, purinyl, 4H-quinolidinyl, quinolinyl, 1,2,3,4-tetrahydroquinolinyl, isoquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 1,8-naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, and any combination thereof.

63. The Ar 1 One or more of H, halo, OH, NO 2 , -SO 3 H, CF 3 , OCF 3 , C 1-6 Alkyl, C 2-6 Alkenyl, C 1-6 Alkoxy, OC 3-4 Alkenyl, -O-benzyl, -O-phenyl, 1,2-methylenedioxy, -NR 5 R 6 , -C(O)OH, -C(O)NH(C 1-6 alkyl), -C(O)NH(C 3-5 alkenyl), -C(O)N(C 1-6 Alkyl)(C 1-6 alkyl), -C(O)(C 3-5 alkenyl)(C 3-6 alkenyl), morpholinyl, piperidinyl, -O-Ar 2 , -CH 2 -(CH 2 ) q -Ar 2 , -O-(CH 2 ) q -Ar 2 , -(CH 2 ) q -O-Ar 2 , or -CH=CH-Ar 2 optionally replaced by; R 5 and R 6 However, independently, H, C 1-6 Alkyl, C 3-6 Alkenyl, C 3-6 selected from the group consisting of alkynyl, and benzyl; Ar 2 is selected from the group consisting of 4-methoxyphenyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, pyrazyl, quinolyl, 3,5-dimethylisoxazolyl, 2-methylthiazolyl, thiazolyl, 2-thienyl, 3-thienyl, and pyrimidinyl; and 62. The compound of claim 61, wherein q is an integer from 0 to 2.

64. R 1 and R 2 61. The compound of claim 60, wherein one or both of are selected from the group consisting of amino acids, β-amino acids, and sugars.

65. R 4 C 1-6 61. The compound of claim 60, which is alkyl.

66. R 4 61. The compound of claim 60, wherein is selected from the group consisting of t-butyl, and 4-{N-[1-(4,4-dimethyl-2,6-dioxocyclohexylidene)-3-methylbutyl]amino}benzyl ester (DMab).

67. The compound of formula II has the structure of formula II-A, II-B, or II-C: 【Transformation 36】 61. The compound of claim 60, wherein the compound has the formula:

68. The following structure: 【Chemistry 37】 A compound having the formula:

69. The following structure: 【Transformation 38】 A compound having the formula:

70. The following structure: 【Chemistry 39】 A compound having the formula:

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