TW201609818A - Macrocyclic inhibitors of the PD-1/PD-L1 and CD80 (B7-1)/PD-L1 protein/protein interactions - Google Patents

Abstract

The present disclosure provides novel macrocyclic peptides which inhibit the PD-1/PD-L1 and PD-L1/CD80 protein/protein interaction, and thus are useful for the amelioration of various diseases, including cancer and infectious diseases.

Description

Macrocyclic inhibitors of PD-1/PD-L1 and CD80(B7-1)/PD-L1 protein/protein interactions

The present invention provides novel macrocyclic peptides that inhibit PD-1/PD-L1 and CD80/PD-L1 protein/protein interactions and are therefore suitable for use in ameliorating various diseases, including cancer and infectious diseases.

The stylized death 1 (PD-1) protein is an inhibitory member of the CD28 receptor family, and the CD28 receptor family also includes CD28, CTLA-4, ICOS, and BTLA. PD-1 is expressed on activated B cells, T cells and bone marrow cells (Agata et al., supra; Okazaki et al, Curr. Opin. Immunol. , 14: 779-782 (2002); Bennett et al. , J. Immunol. , 170: 711-718 (2003)).

The PD-1 protein is a type I 55 kDa transmembrane protein which is part of the Ig gene superfamily (Agata et al., Int. Immunol. , 8: 765-772 (1996)). PD-1 contains a membrane proximal immunoreceptor tyrosine inhibitory element (ITIM) and a tyrosine-based conversion motif (ITSM) at the distal end of the membrane (Thomas, ML, J. Exp. Med. , 181:1953) -1956 (1995); Vivier, E. et al., Immunol . Today, 18: 286-291 (1997)). Although PD-1 is structurally similar to CTLA-4, it lacks the MYPPY motif that is critical for CD80 CD86 (B7-2) binding. Two ligands for PD-1 have been identified: PD-L1 (B7-H1) and PD-L2 (b7-DC). Activation of T cells expressing PD-1 has been shown to be down-regulated when interacting with cells expressing PD-L1 or PD-L2 (Freeman et al, J. Exp. Med. , 192: 1027-1034 (2000); Latchman et al. Human, Nat. Immunol. , 2: 261-268 (2001); Carter et al, Eur. J. Immunol. , 32: 634-643 (2002)). PD-L1 and PD-L2 are members of the B7 protein family that bind to PD-1 but not to other CD28 family members. PD-L1 ligands are abundant in a variety of human cancers (Dong et al., Nat. Med., 8: 787-789 (2002)). The interaction between PD-1 and PD-L1 reduces tumor infiltrating lymphocytes, reduces T cell receptor-mediated proliferation, and immune evasion of cancer cells (Dong et al, J. Mol. Med. , 81:281- 287 (2003); Blank et al, Cancer Immunol. Immunother . , 54: 307-314 (2005); Konishi et al, Clin. Cancer Res. , 10: 5094-5100 (2004)). Immune suppression can be reversed by inhibiting the local interaction between PD-1 and PD-L1, and this reversal is additive when the interaction between PD-1 and PD-L2 is also blocked (Iwai et al. , Proc. Natl. Acad. Sci. USA , 99: 12293-12297 (2002); Brown et al, J. Immunol. , 170: 1257-1266 (2003)).

PD-L1 has also been shown to interact with CD80 (Butte MJ et al., Immunity; 27: 111-122 (2007)). The PD-L1/CD80 interaction that occurs on the expression of immune cells has been shown to be inhibitory. Blocking this interaction has been shown to eliminate this inhibitory interaction (Paterson AM et al, J Immunol. , 187: 1097-1105 (2011); Yang J. et al, J Immunol. , 187(3): 1113- 1119 (August 1, 2011)).

When T cells expressing PD-1 are exposed to cells expressing their ligands, the functional activities (including proliferation, cytokine secretion, and cytotoxicity) in response to antigenic stimulation are reduced. PD-1/PD-L1 or PD-L2 interaction down-regulates immune response during infection or tumor regression, or during self-tolerance development (Keir, ME et al, Annu. Rev. Immunol. , 26: Epub) (2008)). Long-term antigenic stimulation, such as long-term antigenic stimulation that occurs during tumor disease or chronic infection, causes T cells to exhibit PD-1 in high amounts and cause T cell dysfunction in response to long-term antigen activity (reviewed by Kim et al., Curr. Opin.Imm. (2010)). This is called "T cell depletion." B cells also exhibit PD-1/PD ligand inhibition and "depletion".

Blocking PD-1/PD-L1 linkages using antibodies against PD-L1 has been shown to repair and potentiate T cell activation in many systems. Patients with advanced cancer benefit from the treatment of monoclonal antibodies against PD-L1 (Brahmer et al, New Engl. J. Med. (2012)). Preclinical animal models of tumors and chronic infections have shown that blocking the PD-1/PD-L1 pathway by monoclonal antibodies enhances the immune response and can resolve tumors or control infection. Anti-tumor immunotherapy using PD-1/PD-L1 blockade enhances therapeutic immune responses against many histologically diverse tumors (Dong, H. et al., "B7-H1 pathway and its role in the evasion of Tumor immunity", J. Mol. Med. , 81(5): 281-287 (2003); Dong, H. et al., "Tumor-associated B7-H1 promotes T-cell apoptosis: a potential mechanism of immune evasion" , Nat.Med. , 8(8): 793-800 (2002)).

Interfering with PD-1/PD-L1 interaction enhances T cell activity in systems with chronic infection. Blocking PD-L1 increases viral clearance and immune repair in mice with chronic lymphocytic choriomeningitis virus infection (Barber, DL, et al., "Restoring function in exhausted CD8 T cells during chronic viral infection" , Nature , 439 (7077): 682-687 (2006)). Humanized mice infected with HIV-1 showed enhanced protection against viremia and viral depletion of CD4+ T cells (Palmer et al, J. Immunol. (2013)). Blocking PD-1/PD-L1 via a monoclonal antibody against PD-L1 repairs HIV patients (Day, Nature (2006); Petrovas, J. Exp. Med. (2006); Trautman, Nature Med. (2006) ; D'Souza, J. Immunol. (2007); Zhang, Blood (2007); Kaufmann, Nature Imm. (2007); Kasu, J. Immunol. (2010); Porichis, Blood (2011)), HCV patients ( Golden-Mason, J. Virol. (2007); Jeung, J. Leuk. Biol. (2007); Urbani, J. Hepatol. (2008); Nakamoto, PLoS Path. (2009); Nakamoto, Gastroenterology (2008) And HBV patients (Boni, J. Virol. (2007); Fisicaro, Gastro. (2010); Fisicaro et al, Gastroenterology (2012); Boni et al, Gastro. (2012); Penna et al, J. Hep. ( 2012); Raziorrough, Hepatology (2009); Liang, World J. Gastro. (2010); Zhang, Gastro. (2008)) In vitro antigen-specific function of T cells.

Blocking PD-L1/CD80 interactions has also been shown to stimulate immunity (Yang J. et al, J Immunol., 187(3): 1113-9 (August 1, 2011)). Immunostimulation caused by blocking PD-L1/CD80 interactions has been shown to be enhanced by a combination of further blocking PD-1/PD-L1 or PD-1/PD-L2 interactions.

The change in immune cell phenotype is assumed to be an important factor in septic shock (Hotchkiss et al., Nat. Rev. Immunol. (2013)). These changes include an increase in the levels of PD-1 and PD-L1 (Guignant et al., Crit. Care (2011)), and increased levels of PD-1 and PD-L1 in cells of patients with septic shock exhibiting increased T cells. The degree of apoptosis. Antibodies against PD-L1 reduce the extent of apoptosis in immune cells (Zhang et al, Crit. Care (2011)). In addition, mice lacking PD-1 expression were more tolerant to septic shock symptoms than wild-type mice (Yang J. et al., J. Immunol., 187(3): 1113-1119 (8, 2011) January 1))). Studies have revealed that the use of antibodies to block PD-L1 interactions can inhibit inappropriate immune responses and improve disease signs.

In addition to enhancing the immune response to long-term antigens, blocking the PD-1/PD-L1 pathway has also been shown to enhance response to vaccination, including therapeutic vaccination in the context of chronic infection (Ha, SJ et al, "Enhancing therapeutic vaccination by blocking PD-1-mediated inhibitory signals during chronic infection", J. Exp. Med. , 205(3): 543-555 (2008); Finnefrock, AC et al., "PD-1 blockade in rhesus Macaques: impact on chronic infection and prophylactic vaccination", J. Immunol. , 182(2): 980-987 (2009); Song, M.-Y. et al., "Enhancement of vaccine-induced primary and memory CD8+ t- Cell responses by soluble PD-1", J. Immunother. , 34(3): 297-306 (2011)).

The molecules described herein both show the ability to block the interaction of PD-L1 with PD-1 in both biochemical and cell-based assay systems. These results are consistent with the potential for therapeutic administration (including therapeutic vaccines) to enhance immunity in cancer or chronic infection.

The macrocyclic peptides described herein inhibit the interaction of PD-L1 with PD-1 and with CD80 use. These compounds have been shown to bind highly efficiently to PD-L1, block the interaction of PD-L1 with PD-1 or CD80, and promote T-cell functional activity, making it parenteral, oral, lung, Candidates for nasal, buccal, and sustained release formulations.

In one embodiment, the invention provides a compound of formula (I) Or a pharmaceutically acceptable salt thereof, wherein: A is selected from the group consisting of a bond, among them: Represents the point of attachment to the carbonyl group and Represents a point of attachment to a nitrogen atom; n is 0 or 1; R ¹⁴ and R ^{15 are} independently selected from hydrogen and methyl; and R ¹⁶ is selected from hydrogen, -CHR ¹⁷ C(O)NH ₂ , -CHR ¹⁷ C (O) NHCHR ¹⁸ C(O)NH ₂ , and -CHR ¹⁷ C(O)NHCHR ¹⁸ C(O)NHCH ₂ C(O)NH ₂ ; wherein R ¹⁷ is selected from hydrogen and -CH ₂ OH and wherein R ¹⁸ is selected from the group consisting of hydrogen and methyl; R ^c , R ^f , R ^h , R ⁱ , R ^m and R ⁿ are hydrogen; R ^a , R ^e , R ^j and R ^{k are} each independently selected from hydrogen and methyl; ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² and R ^{13 are} independently selected from the group consisting of a natural amino acid side chain and a non-natural amine a side chain of a base acid, or a ring formed with a corresponding ortho R group as described below; each of R ^e and R ^k and the corresponding ortho R group and the atom to which they are attached form an azetidine, pyrrolidine, a ring of morpholine, piperidine, piperazine, and tetrahydrothiazole; wherein each ring is optionally substituted with one to four groups independently selected from the group consisting of an amine group, a cyano group, a methyl group, a halogen group, and a hydroxyl group; and R ^b is a methyl group. or R ^b and R ² form together with the atoms to which they are attached are selected from azetidine, pyrrolidine, morpholine, piperidine, piperazine and thiazolidine ring; wherein each surveying Group is independently selected conditions, cyano, methyl, halo and hydroxy radicals substituted with one to four; R ^d is hydrogen or methyl, or R ^d and R ⁴ may form together with the atoms to which they are attached are selected from the a ring of azetidine, pyrrolidine, morpholine, piperidine, piperazine, and tetrahydrothiazole; wherein each ring is optionally one to four selected from the group consisting of an amine group, a cyano group, a methyl group, a halogen group, a hydroxyl group, and a phenyl group. a group substituted; R ^g is hydrogen or methyl, or R ^g and R ⁷ together with the atom to which they are attached form a ring selected from the group consisting of azetidine, pyrrolidine, morpholine, piperidine, piperazine, and tetrahydrothiazole; Wherein each ring is substituted with one to four groups independently selected from the group consisting of: an amine group, a benzyl group substituted with a halogen group, a benzyloxy group, a cyano group, a cyclohexyl group, a methyl group, a halogen group, a hydroxyl group, a quinolinyloxy group optionally substituted by a methoxy group, a quinolinyloxy group optionally substituted by a halogen group, and a tetrazolyl group, wherein the pyrrolidine and piperidine are optionally cyclic with a cyclohexyl group, a phenyl group or a fluorene group. fused group; and R ¹ is methyl, or R ¹ and R ¹² is selected form azetidine and pyrrolidine ring together with the atoms which they are attached, wherein each ring Substituted with one to four substituents independently selected from amino, cyano, methyl, halo, and hydroxy groups.

In another embodiment, the invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, wherein A is

In another embodiment, the invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, wherein A is R ^d is a methyl group, or R ^d and R ⁴ together with the atom to which they are attached form a ring selected from azetidine, pyrrolidine, morpholine, piperidine, piperazine and tetrahydrothiazole; Two groups independently selected from the group consisting of an amino group, a cyano group, a methyl group, a halogen group, a hydroxyl group, and a phenyl group; R ^g is a methyl group, or R ^g and R ⁷ together with the atom to which they are attached form an azetidine group a ring of pyrrolidine, morpholine, piperidine, piperazine and tetrahydrothiazole; wherein each ring is optionally substituted with one or two groups independently selected from the group consisting of an amine group and, optionally, a benzyl group substituted by a halogen group. a benzyloxy group, a cyano group, a cyclohexyl group, a methyl group, a halogen group, a hydroxyl group, an isoquinolineoxy group optionally substituted by a methoxy group, a quinolineoxy group optionally substituted by a halogen group, and a tetrazolyl group And wherein the pyrrolidine and piperidine are cyclized with a cyclohexyl, phenyl or hydrazine group; and R ^k is a methyl group, or R ^k and R ¹¹ together with the atom to which they are attached form an azetidine a ring of pyrrolidine, morpholine, piperidine, piperazine and tetrahydrothiazole; wherein each ring is independently selected from the group consisting of an amine group, a cyano group, a methyl group, and a halogen group. And a hydroxy group substitution; in another embodiment, the invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, wherein A is R ^d and R ⁴ together with the atom to which they are attached form a ring selected from the group consisting of azetidine, pyrrolidine, morpholine, piperidine, piperazine and tetrahydrothiazole; wherein each ring is optionally independently selected from an amine group by one or two Substituting a group of cyano, methyl, halo and hydroxy; R ^g and R ⁷ together with the atom to which they are attached form a pyrrolidine ring wherein the ring is optionally substituted with one or two groups independently selected from: Amine, optionally substituted by halo, benzyl, benzyloxy, cyano, cyclohexyl, methyl, halo, hydroxy, optionally methoxy-substituted quinolinyloxy, optionally a halo substituted quinolinyloxy group, and a tetrazolyl group; and wherein the pyrrolidine and piperidine are optionally fused to a cyclohexyl, phenyl or hydrazine group; and R ^k is a methyl group.

In another embodiment, the invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, wherein A is R ^d and R ⁴ together with the atom to which they are attached form a ring selected from the group consisting of azetidine, pyrrolidine, morpholine, piperidine, piperazine and tetrahydrothiazole; wherein each ring is optionally independently selected from an amine group by one or two Substituting a group of cyano, methyl, halo and hydroxy; R ^g and R ⁷ together with the atom to which they are attached form a pyrrolidine ring wherein the ring is optionally substituted with one or two groups independently selected from: Amine, optionally substituted by halo, benzyl, benzyloxy, cyano, cyclohexyl, methyl, halo, hydroxy, optionally methoxy-substituted quinolinyloxy, optionally a halo substituted quinolinyloxy group, and a tetrazolyl group; and wherein the pyrrolidine and piperidine are optionally fused to a cyclohexyl, phenyl or hydrazine group; R ^k is a methyl group; and R ⁸ is selected from the group consisting of Azaindenyl C ₁ -C ₃ alkyl, benzothiazolyl C ₁ -C ₃ alkyl, benzothienyl C ₁ -C ₃ alkyl, benzyloxy C ₁ -C ₃ alkyl, Diphenylmethyl, furyl C ₁ -C ₃ alkyl, imidazolyl C ₁ -C ₃ alkyl, naphthyl C ₁ -C ₃ alkyl, pyridyl C ₁ -C ₃ alkyl, thiazolyl C ₁ -C ₃ alkyl group, thienyl C ₁ -C ₃ alkyl; and Indolyl C ₁ -C ₃ alkyl, wherein the indole moiety is optionally substituted with one substituent selected from C ₁ -C ₃ alkyl, cyano, halo and hydroxy group.

In another embodiment, the invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, wherein A is R ^d and R ⁴ together with the atom to which they are attached form a ring selected from the group consisting of azetidine, pyrrolidine, morpholine, piperidine, piperazine and tetrahydrothiazole; wherein each ring is optionally independently selected from an amine group by one or two Substituting a group of cyano, methyl, halo and hydroxy; R ^g and R ⁷ together with the atom to which they are attached form a pyrrolidine ring wherein the ring is optionally substituted with one or two groups independently selected from: Amine, optionally substituted by halo, benzyl, benzyloxy, cyano, cyclohexyl, methyl, halo, hydroxy, optionally methoxy-substituted quinolinyloxy, optionally a halo substituted quinolinyloxy group, and a tetrazolyl group; and wherein the pyrrolidine and piperidine ring are fused to a cyclohexyl, phenyl or hydrazine group; R ^k is a methyl group; and R ⁸ is 3- Mercapto C ₁ -C ₃ alkyl, optionally substituted with a group selected from C ₁ -C ₃ alkyl, halo, hydroxy or cyano.

In another embodiment, the invention provides a compound of formula (II) Or a pharmaceutically acceptable salt thereof, wherein: A is selected from the group consisting of a bond, among them: Represents the point of attachment to the carbonyl group and Represents a point of attachment to a nitrogen atom; n is 0 or 1; R ¹⁴ and R ^{15 are} independently selected from hydrogen and methyl; and R ¹⁶ is selected from hydrogen, -CHR ¹⁷ C(O)NH ₂ , -CHR ¹⁷ C (O) NHCHR ¹⁸ C(O)NH ₂ , and -CHR ¹⁷ C(O)NHCHR ¹⁸ C(O)NHCH ₂ C(O)NH ₂ ; wherein R ¹⁷ is selected from hydrogen and -CH ₂ OH and wherein R ¹⁸ is selected from hydrogen and ^{methyl; R a, R f, R} j, R k, R l and R ^m is hydrogen; R ^b and R ^c is methyl; R ^g is selected from hydrogen and methyl; R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ and R ^{12 are} independently selected from the group consisting of a natural amino acid side chain and an unnatural amino acid side chain Or forming a ring with the corresponding ortho-R group as described below; R ^d is selected from hydrogen or methyl, or R ^d and R ⁴ together with the atom to which they are attached form an azetidine, pyrrolidine, morpholine a ring of piperidine, piperazine and tetrahydrothiazole; wherein each ring is optionally substituted with one to four groups independently selected from the group consisting of an amine group, a cyano group, a methyl group, a halomethyl group and a hydroxyl group; and the R ^e is selected from the group consisting of hydrogen and forming methyl, or R ^e and R ⁵ together with the atom to which they are attached are selected from azetidine, pyrrolidine, morpholine, piperidine, piperazine and thiazolidine ring; wherein each Optionally independently selected from amino, cyano, methyl, halo, halomethyl, and a hydroxyl group substituted by one to four; R ^h is selected from hydrogen and methyl, or R ^h and R ⁸ connected to it The atoms together form a ring selected from the group consisting of azetidine, pyrrolidine, morpholine, piperidine, piperazine, and tetrahydrothiazole; wherein each ring is optionally independently selected from the group consisting of an amine group, a cyano group, a methyl group, and a halogen group. a group of a halomethyl group and a hydroxy group; and R ⁱ is selected from the group consisting of hydrogen and methyl, or R ⁱ and R ⁹ together with the atom to which they are attached form an azetidine, pyrrolidine, morpholine, piperidine, a ring of piperazine and tetrahydrothiazole; wherein each ring is optionally substituted with one to four groups independently selected from the group consisting of an amino group, a cyano group, a methyl group, a halogen group, a halomethyl group, and a hydroxyl group. In another embodiment, the invention provides a compound of formula (II), or a pharmaceutically acceptable salt thereof, wherein A is

In another embodiment, the invention provides a compound of formula (II), or a pharmaceutically acceptable salt thereof, wherein A is R ^d is a methyl group, or R ^d and R ⁴ together with the atom to which they are attached form a ring selected from azetidine, pyrrolidine, morpholine, piperidine, piperazine and tetrahydrothiazole; Two groups independently selected from the group consisting of an amino group, a cyano group, a methyl group, a halomethyl group and a hydroxyl group; R ^g is a methyl group; and; R ⁱ is a methyl group, or R ⁱ and R ⁹ together with the atom to which they are attached Forming a ring selected from the group consisting of azetidine, pyrrolidine, morpholine, piperidine, piperazine, and tetrahydrothiazole; wherein each ring is independently selected from the group consisting of an amine group, a cyano group, a methyl group, a halogen group, and a halogen. Substituents of methyl and hydroxy groups are substituted.

In another embodiment, the invention provides a compound of formula (II), or a pharmaceutically acceptable salt thereof, wherein A is R ^d is a methyl group, or R ^d and R ⁴ together with the atom to which they are attached form a ring selected from azetidine, pyrrolidine, morpholine, piperidine, piperazine and tetrahydrothiazole; Two groups independently selected from the group consisting of amino, cyano, methyl, halomethyl and hydroxy; R ^g is methyl; R ⁱ is methyl, or R ⁱ and R ⁹ are taken together with the atom to which they are attached a ring of aziridine, pyrrolidine, morpholine, piperidine, piperazine and tetrahydrothiazole; wherein each ring is independently selected from the group consisting of an amine group, a cyano group, a methyl group, a halogen group, and a halomethyl group. And a group substituted with a hydroxyl group; and R ⁷ is a phenyl C ₁ -C ₃ alkyl group, optionally substituted by a fluorine group.

In another embodiment, the invention provides a method of enhancing, stimulating, and/or enhancing an immune response in an individual in need thereof, the method comprising administering to the individual a therapeutically effective amount of at least one macrocyclic peptide described herein. In another embodiment, the method further comprises administering another agent before, after or simultaneously with administration of the macrocyclic peptide or peptide described herein. In another embodiment, the other agent is an antimicrobial, an antiviral, a cytotoxic agent, and/or an immune response modifier.

In another embodiment, the invention provides a method of inhibiting growth, proliferation or metastasis of cancer cells in an individual in need thereof, the method comprising administering to the individual a therapeutically effective amount of one or more of the macrocyclic peptides described herein. In another embodiment, the cancer is selected from the group consisting of melanoma, renal cell carcinoma, squamous non-small cell lung cancer (NSCLC), non-squamous NSCLC, colorectal cancer, castration-refractory prostate cancer, ovarian cancer, gastric cancer, and hepatocytes. Cancer, pancreatic cancer, head and neck squamous cell carcinoma, esophageal cancer, gastrointestinal cancer and breast cancer, and hematological malignancies.

In another embodiment, the invention provides a method of treating an infectious disease in an individual in need thereof, the method comprising administering to the individual a therapeutically effective amount of at least one macrocyclic peptide described herein. In another embodiment, the infectious disease is caused by a virus. In another embodiment, the virus is selected from the group consisting of HIV, hepatitis A, hepatitis B, hepatitis C, herpes virus, and influenza.

In another embodiment, the invention provides a method of treating septic shock in an individual in need thereof, the method comprising administering to the individual a therapeutically effective amount of one or more of the macrocyclic peptides described herein.

In another embodiment, the invention provides a method of blocking the interaction of PD-L1 with PD-1 and/or CD80 in an individual, the method comprising administering to the individual at least one of the therapeutically effective amounts described herein Macrocyclic peptide.

In compounds of formula (I) and (II) wherein the R side chain is part of a methyl substituted ring, it is understood that the methyl group may be in the ring of any substitutable carbon atom (including as part of a macrocyclic parent structure) On the carbon).

In the compound of formula (I), the R ¹ side chain is preferably: phenylalanine, tyrosine, 3-thiophen-2-yl, 4-methylphenylalanine, 4-chlorophenylalanine, 3-methoxy amphetamine Acid, isochryonic acid, 3-methylphenylalanine, 1-naphthylalanine, 3,4-difluorophenylalanine, 4-fluorophenylalanine, 3,4-dimethoxyphenylalanine, 3,4- Dichlorophenylalanine, 4-difluoromethylphenylalanine, 2-methylphenylalanine, 2-naphthylalanine, tryptophan, 4-pyridyl, 4-bromophenylalanine, 3-pyridyl, 4- Trifluoromethylphenylalanine, 4-carboxyphenylalanine, 4-methoxyphenylalanine, biphenylalanine, and 3-chlorophenylalanine; and 2,4-diaminobutane.

In the compound of the formula (I) wherein R ^{2 is} not a part of the ring, the R ² side chain is preferably: alanine, serine and glycine.

In the compound of formula (I), the R ³ side chain is preferably: aspartic acid, aspartic acid, glutamic acid, glutamic acid, serine, ornithine, lysine, histamine Acid, sulphate, leucine, alanine, 2,3-diaminopropane and 2,4-diaminobutane.

In the compound of the formula (I) wherein R ^{4 is} not a part of the ring, the R ⁴ side chain is preferably: valine acid, alanine, isoleucine and glycine.

In the compound of formula (I), the R ⁵ side chain is preferably: histidine, aspartic acid, 2,3-diaminopropane, serine, glycine, 2,4-diamine Butane, sulphate, alanine, lysine, aspartic acid, alanine and 3-thiazolyl alanine.

In the compound of formula (I), the R ⁶ side chain is preferably: leucine, aspartic acid, aspartic acid, glutamic acid, glutamic acid, serine, lysine, 3- Cyclohexane, sulphate, ornithine, 2,4-diaminobutane, alanine, arginine and ornithine (COCH ₃ ).

In the compound of the formula (I) wherein R ^{7 is} not a part of the ring, the R ⁷ side chain is preferably glycine, 2,4-diaminobutane, serine, lysine, arginine, Amino acid, histidine, aspartic acid, glutamic acid, alanine and 2,4-diaminobutane (C(O)cyclobutane).

In the compound of formula (I), the R ⁸ side chain is preferably tryptophan and 1,2-benzisothiazolinylalanine.

In the compound of formula (I), the R ⁹ side chain is preferably: serine, histidine, lysine, ornithine, 2,4-dibutylamine, threonine, lysine, glycine Amine acid, glutamic acid, valine acid, 2,3-diaminopropane, arginine, aspartic acid and tyrosine.

In the compound of formula (I), the R ¹⁰ side chain is preferably tryptophan, benzisothiazolyl alanine, 1-naphthylalanine, 5-fluorotryptophan, methionine, 7-methyl. Tryptophan, 5-chlorotryptamine and -methyltryptophan.

In the compound of formula (I), the R ¹¹ side chain is preferably: orthanoic acid, leucine, aspartic acid, phenylalanine, methionine, ethoxymethane, alanine, tryptophan. , isoleucine, phenylpropane, glutamic acid, hexane and heptane.

In the compound of the formula (I) wherein R ^{12 is} not a part of the ring, the R ¹² side chain is preferably: orthanoic acid, alanine, ethoxymethane, methionine, serine, phenylalanine, Methoxyethane, leucine, tryptophan, isoleucine, glutamic acid, hexane, heptane and glycine.

In the compound of formula (I), the R ¹³ side chain is preferably: arginine, ornithine, alanine, 2,4-diaminobutane, 2,3-diaminopropane, leucine, Aspartic acid, glutamic acid, serine, lysine, threonine, cyclopropylmethane, glycine, valine, isoleucine, histidine and 2-aminobutane.

In the compound of the formula (II), the R ¹ side chain is preferably: phenylalanine, 3-methoxyphenylalanine, 2-fluorophenylalanine, 3-fluorophenylalanine, 4-fluorophenylalanine, 3,4-di Fluoroamphetamine, 3,5-difluorophenylalanine, 3,4,5-trifluorophenylalanine, 3-fluoro, 4-chlorophenylalanine, 3-chloro, 4-fluorophenylalanine, 3-chlorophenylalanine, 4-chlorophenylalanine, 3,4-dichlorophenylalanine, 3,5-dichlorophenylalanine, 3,5-dichloro, 4-fluorophenylalanine, 3-chloro, 4,5-difluorophenylalanine, 4-bromophenylalanine, 4-nitrophenylalanine, 3-trifluoromethylphenylalanine, 4-trifluoromethylphenylalanine and 3-pyridylalanine.

In the compound of formula (II), the R ² side chain is preferably: phenylalanine, alanine, histidine, tyrosine, tryptophan, glutamic acid, 1-naphthylalanine, 2-naphthylpropylamine Acid, 2-benzothiazolyl alanine, 3-pyridyl alanine and 4-pyridyl alanine.

In the compounds of formula (II), R ³ is the side chain is preferably: n-leucine, alanine, tyrosine, glutamic acid, leucine and isoleucine.

In the compound of the formula (II) wherein R ^{4 is} not a part of the ring, the R ⁴ side chain is preferably: glycine and alanine.

In the compound of the formula (II) wherein R ^{5 is} not a part of the ring, the R ⁵ side chain is preferably: aspartic acid, glutamic acid, arginine, lysine, aspartic acid, silkamine Acid, 2,4-diaminobutane, 2,3-diaminopropane and 2-aminobutane.

In the compound of the formula (II), the R ⁶ side chain is preferably: valine acid, leucine, isoleucine, N-methylthreonate and cyclohexylmethane.

In the compound of the formula (II), the R ⁷ side chain is preferably: phenylalanine and 3-fluorophenylalanine.

In the compound of the formula (II) wherein R ^{8 is} not a part of the ring, the R ⁸ side chain is preferably: tyrosine, 3-iodotyrosine, leucine, arginine, glutamic acid, bran Amino acid, pentafluorophenylalanine, 4-aminophenylalanine, 4-aminomethylphenylalanine, 3,4-dimethoxyphenylalanine, tryptophan, 5-chlorotryptophan, 5-hydroxyl Amine acid, isochryamic acid, lysine, ornithine and 2,3-diaminopropane.

In the compounds of formula (II) is, R ¹⁰ is preferably the side chain of: tryptophan, 5-chloro-tryptophan, 7-aza-tryptophan, leucine Leis, 3-benzothiazolyl, and 1-alanine Naphthylalanine.

In the compound of the formula (II), the R ¹¹ side chain is preferably tyrosine, 4-fluorophenylalanine, 4-aminomethylphenylalanine, 4-aminophenylalanine and 3,4-dihydroxyphenylalanine.

In the compound of formula (II), the R ¹² side chain is preferably: leucine, tyrosine, arginine, lysine, ornithine, glutamic acid, phenylalanine, 4-methylphenylalanine, 4-chlorophenylalanine, 4-aminomethylphenylalanine, norleucine, cyclohexylalanine, 2,4-diaminobutane and 2,3-diaminopropane.

In the compound of the formula (II), when R ⁴ and R ⁹ are a part of the ring, the preferred stereochemical configuration is the D-isomer, and when R ⁵ and R ⁸ are a part of the ring, preferred stereochemistry The configuration is an L-isomer.

One embodiment of the subject matter described herein is directed to a polypeptide comprising a sequence of Formula I(a):

Wherein: A is an organic or peptide linker between X _aa1 and X _aa14 to give a macrocyclic peptide; X _aa1 is a natural or non-naturally occurring aromatic or heteroaromatic or alkyl or heteroarylalkylamine group Acid; X _aa2 is a natural or non-naturally occurring alkyl or N-methylated alkyl amino acid; X _aa3 is a natural or non-naturally occurring hydrophilic or alkyl or polar amino acid; X _aa4 is natural or a non-naturally occurring amino acid, an alkyl amino acid or an N-methylated alkyl amino acid; X _aa5 is a naturally or non-naturally occurring heteroaromatic amino acid or a positively charged amino acid or alkane Amino acid; X _aa6 is a natural or non-naturally occurring hydrophilic or hydrophobic or positively or negatively charged amino acid; X _aa7 is a natural or non-naturally occurring N-methylated or non-N- Methylated hydrophilic or hydrophobic or positively or negatively charged amino acids; X _aa8 is a natural or non-naturally occurring aromatic or heteroaromatic or arylalkyl or heteroarylalkylamino acid ; X _aa9 is a naturally or non-naturally occurring hydrophilic or the hydrophobic or positively charged or negatively charged amino _acid; X aa10 is a naturally or non-naturally occurring aromatic or heteroaromatic of Or arylalkyl or heteroarylalkyl or heteroalkyl, or alkyl amino acid; X _aa11 is a naturally or non-naturally occurring or non-methylation of N- methyl-N- or alkyl or heteroalkyl An aromatic or heteroaromatic amino acid; X _aa12 is a naturally or non-naturally occurring N-methylated or non-N-methylated alkyl or heteroalkyl or aromatic or heteroaromatic amino acid; X _aa13 a naturally or non-naturally occurring hydrophilic or hydrophobic or positively or negatively charged amino acid; X _{aa 14} is a natural or non-naturally occurring amino acid having a suitable activation with linker A A functional group that reacts at one end to produce a cyclic peptide; X _aa15 is a naturally occurring or non-naturally occurring amino acid or spacer followed by a label or spacer followed by a solubilizing element or a PK enhancing element.

In another embodiment, the subject matter described herein pertains to a polypeptide comprising a sequence of Formula I(b):

Wherein: A is an electrophilic moiety, such as a Michael acceptor or a chloroethyl or bromoethyl group, which is capable of _reacting with a sulfhydryl group present on residue X _{aa 14} to form a covalent thioether bond, Thereby producing a macrocyclic peptide; wherein the thioether bond may or may not be oxidized to the corresponding diastereomeric hydrazide; and wherein A may optionally be present; and wherein if A is present, it may be Gly or other having a free amine a spacer which can be used for cyclization of a peptide via a _guanamine bond with a carboxyl group on a side chain of X _aa14 to obtain a cyclic peptide having an N-terminal end to a _{guanamine in} the side chain; and wherein if A is not present, The N-terminal amino group of the X _aa1 residue can be used for cyclization of the peptide via a _guanamine bond with a carboxyl group on the side chain of X _{aa 14} to obtain a cyclic peptide having an N-terminal end to the indole amine in the side chain; _Where A is present, it may be Gly or other spacer having a free amine which can be used for the cyclization of the peptide via a _{guanamine bond with} the C-terminal α-carboxyl group of X _aa15 to give an end-to-end cyclic peptide; when A is absent, then X _aa1 amino acid N-terminal acyl group may be formed via a C-terminus of _X aa15 carboxy α- The peptide bonds of the cyclized end to end to give the peptide chain; X _aa1 is a naturally or non-naturally occurring amino acid of which contains L-Phe, L-Ala, L-Trp, L-Tyr, L-Phe (4-OMe), L-Phe(4-F), L-Phe(4-Cl), L-Phe(4-Br), L-Phe(4-Me), L-Phe(4-CF ₃ ), L-Phe(4- t- Bu), L-Phe(5-F), L-1-Nal, L-2-Nal, L-Bip, L- ^m Phe, L-Tic, L-3 -Pya, L-4-Pya, L-Tza, L-3-Tha; X _aa2 is a natural or non-naturally occurring amino acid selected from the group consisting of L-Ala, L- ^m Ala, ^m Gly, L- ^m a group consisting of Val; X _aa3 is selected from the group consisting of Gly, L-Asn and L-Ala; X _aa4 is a natural or non-naturally occurring amino acid comprising L-Pro, L-Ala, L-α- Me-Pro, L-Pro (4R-OH), L-Pro (4R-OBzl), L-Pro (4R-NH ₂ ), L-Pro (3R-Ph), L-Pro (4S-Ph), L-Pro(5R-Ph), L-Azt, L-Pip, L-Oic, L-2, 3-A bridge-Pro, L-3, 4-A bridge-Pro, L-Val, L-Leu , L-Ile, L- ^m Ala, L- ^m Val, L- ^m Leu, L-Tza; X _aa5 is a natural or non-naturally occurring amino acid selected from the group consisting of L-His, L -Ala, L-Tza, L-Arg, L-Lys, L-Orn, L-Dab and L-Dap; X _aa6 is naturally or non-naturally occurring An amino acid comprising L-Leu, L-Ala, L-Arg, L-His, L-Glu and L-Asp; X _aa7 is a natural or non-naturally occurring amino acid comprising ^m Gly, Gly, L- ^m Ala, D- ^m Ala, L-Pro, L-Ser, L- ^m Ser L-Dab, L-Arg and L-His; X _aa8 is L-Trp, L-Phe, L-Tyr, L -His, L-Phe(5-F), L-Tza, L-Bzt, L-1-Nal, L-2-Nal, L-2-Pya, L-3-Pya, L-4-Pya; X _aa9 is a natural or non-naturally occurring amino acid comprising L-Ser, L-Ala, L-Arg and D-Asn; X _aa10 is a natural or non-naturally occurring amino acid selected from the group consisting of Group: L-Trp, L-Ala, L-Met, L-Nle, L-Leu and L-Ile, L-Phe, L-Tyr, L-His, L-Phe (5-F), L-Tza , L-Bzt, L-1-Nal, L-2-Nal, L-2-Pya, L-3-Pya, L-4-Pya; X _aa11 is a natural or non-naturally occurring amino acid, which comprises L- ^m Nle, L-Nle, L- ^m Ala, L-Ala, L-Phe, L- ^m Phe and L- ^m Leu, L-Ser, D-Nle and L-Pro; X _aa12 is natural or _non- a naturally occurring amino acid comprising L- ^m Nle, L-Nle, L- ^m Ala, L-Ala, L-Phe, L- ^m Phe, L- ^m Leu and L-Pro; X _aa13 is natural or a non-naturally occurring amino acid comprising L-Arg, LA La, L-Leu, L-Lys, L-Asp, L-Glu, L-His; X _aa14 is selected from the group consisting of L-Cys, D-Cys, Asp, Glu, Gly, L-high- Cys, D-high-Cys, L-Pen, D-Pen, L- ^m Cys, and D- ^m Cys; X _aa15 is Gly or Gly, followed by a PEG spacer comprising at least two ethylene glycol units, or Gly, followed by a PEG spacer comprising at least two ethylene glycol units, followed by a label such as biotin, or Gly, followed by a spacer followed by a PK enhancing element; wherein X _aa15 is present and Wherein the C-terminal carbonyl carbon of the amino acid is attached to a hydroxyl group to form a carboxylic acid or is attached to a nitrogen to form a carboxamide (NH ₂ ), an alkyl carboxamide (NHR ¹ ) or a dialkyl carboxamide (NR ¹ R ² ); wherein R ¹ and R ^{2 are} each alkyl or arylalkyl; wherein if X _{aa 15} is absent, the C-terminal carbonyl carbon of X _{aa 14} is bonded to nitrogen to form carboxamide (NH ₂ ), alkyl Carboxyguanamine (NHR ₁ ) or dialkyl carboxamide (NR ₁ R ₂ ); wherein each of R ₁ and R ₂ is an alkyl or arylalkyl group.

In another embodiment, the subject matter described herein pertains to a polypeptide comprising a sequence of Formula I(c):

Wherein: A is an electrophilic moiety, such as a Michael acceptor or a chloroethyl or bromoethyl group, which is capable of _reacting with a sulfhydryl group present on residue X _{aa 14} to form a covalent thioether bond, Thereby producing a macrocyclic peptide; wherein the thioether bond may or may not be oxidized to the corresponding diastereomeric hydrazide; and wherein A may optionally be present; and wherein if A is present, it may be Gly or other having a free amine a spacer which can be used for cyclization of a peptide via a _guanamine bond with a carboxyl group on a side chain of X _aa14 to obtain a cyclic peptide having an N-terminal end to a _{guanamine in} the side chain; and wherein if A is not present, The N-terminal amino group of the X _aa1 residue can be used for cyclization of the peptide via a _guanamine bond with a carboxyl group on the side chain of X _aa14 to obtain a cyclic peptide having an N-terminal end to a _{guanamine in} the side chain; _Where A is present, it may be Gly or other spacer having a free amine which can be used for the cyclization of the peptide via a _{guanamine bond with} the C-terminal α-carboxyl group of X _aa15 to give an end-to-end cyclic peptide; If A is absent, the N-terminal amine group of the X _aa1 amino acid can form a _guanamine via the C-terminal α-carboxyl group of X _aa15 The bond is used for cyclization of the peptide to obtain an end-to-end cyclic peptide; X _aa1 is a natural or non-naturally occurring amino acid comprising L-Phe, L-Ala, L-Trp, L-Tyr, L-Phe (4-OMe), L-Phe(4-F), L-Phe(4-Cl), L-Phe(4-Br), L-Phe(4-Me), L-Phe(4-CF ₃ ), L-Phe(4-t-Bu), L-Phe(5-F), L-1-Nal, L-2-Nal, L-Bip, L- ^m Phe, L-Tic, L-3 -Pya, L-4-Pya, L-Tza, L-3-Tha; X _aa2 is a natural or non-naturally occurring amino acid selected from the group consisting of L-Ala, L- ^m Ala, ^m Gly, L- ^m a group consisting of Val; X _aa3 is selected from the group consisting of Gly, L-Asn and L-Ala; X _aa4 is a natural or non-naturally occurring amino acid comprising L-Pro, L-Ala, L-α- Me-Pro, L-Pro (4R-OH), L-Pro (4R-NH ₂ ), L-Pro (4S-Ph), L-Azt, L-Pip and L-Oic; X _aa5 is natural or _non- a naturally occurring amino acid selected from the group consisting of L-His and L-Ala; X _aa6 is a natural or non-naturally occurring amino acid comprising L-Leu, L-Ala, L-Arg and L- Asp; X _aa7 is a natural or non-naturally occurring amino acid comprising ^m Gly, Gly, L- ^m Ala, D- ^m Ala, L-Pro, L-Ser, L- ^m Ser L-Dab, L- Arg and L-His; X _aa9 is a naturally The non-naturally occurring amino acid, which comprises L-Ser, L-Ala, L-Arg and _D-Asn; X aa10 is a naturally or non-naturally occurring amino acids of, selected from the group consisting of: L-Trp , L-Ala, L-Met, L-Leu and L-Ile; X _aa11 is a natural or non-naturally occurring amino acid comprising L- ^m Nle, L-Nle, L- ^m Ala, L-Ala, L-Phe, L- ^m Phe and L- ^m Leu, L-Ser and D-Nle; X _aa12 is a natural or non-naturally occurring amino acid comprising L- ^m Nle, L-Nle, L- ^m Ala And L-Ala; X _aa13 is a natural or non-naturally occurring amino acid comprising L-Arg, L-Ala and L-Leu; X _aa14 is selected from the group consisting of L-Cys, D-Cys, Asp, Glu and Gly a group of components; X _aa15 is Gly; or Gly, followed by a PEG spacer comprising at least two ethylene glycol units; or Gly, followed by a PEG spacer comprising at least two ethylene glycol units, followed by a label, such as biotin; wherein X _{aa15 is} optionally present and wherein the C-terminal carbonyl carbon of the amino acid is attached to the nitrogen to form a carboxamide (NH ₂ ), an alkyl carboxamide (NHR ₁ ) or a dialkyl carboxylate Amides (NR ₁ R _2); wherein R ₁ and R ₂ are each alkyl or aryl group; wherein if _X aa15 absent Then X C-terminal carbonyl carbon attached to the nitrogen of _aa14 is formed 2carboxamide (NH _2), alkyl 2carboxamide (NHR ₁₎ or dialkyl 2carboxamide (NR ₁ R _2); wherein R ₁ and R ₂ Each is an alkyl or arylalkyl group.

In another embodiment, the subject matter described herein pertains to a polypeptide comprising a sequence of Formula II (a):

Wherein: A is an organic or peptide linker between X _aa1 and X _aa13 to give a macrocyclic peptide; X _aa1 is a natural or non-naturally occurring aromatic or heteroaromatic or alkyl or heteroarylalkylamine group Acid; X _aa2 is a natural or non-naturally occurring alkyl or aromatic N-methylated amino acid; X _aa3 is a natural or non-naturally occurring hydrophobic N-methylated amino acid; X _aa4 is natural or Non-naturally occurring hydrophobic N-methylated amino acids; X _aa5 is a natural or non-naturally occurring alkyl amino acid or a positively or negatively charged amino acid; X _aa6 is naturally or non-naturally occurring a hydrophobic amino acid; X _aa7 is a naturally or non-naturally occurring N-methylated or non-N-methylated aromatic or heteroaromatic or alkyl or heteroarylalkylamino acid; X _aa8 is A natural or non-naturally occurring aromatic or heteroaromatic or arylalkyl or heteroarylalkylamino acid or alkylamino acid; X _aa9 is a N-methylated or non-N naturally or non-naturally occurring a methylated aromatic or heteroaromatic or alkyl or heteroarylalkylamino acid; X _aa10 is a natural or non-naturally occurring aromatic or heteroaromatic or arylalkyl or heteroarylalkyl group or Alkyl or heteroalkyl Acid; X _aa11 is a naturally or non-naturally occurring, or of aromatic or heteroaromatic arylalkyl or heteroarylalkyl or heteroalkyl, or alkyl amino acid; X _aa12 is a naturally or non-naturally occurring Fang a family or heteroaromatic or arylalkyl or heteroarylalkyl or alkyl or heteroalkylamino acid; X _aa13 is a naturally occurring or non-naturally occurring amino acid having a suitable activation with a linker The functional group reactive at one end of A to produce a cyclic peptide; X _aa14 is a naturally occurring or non-naturally occurring amino acid or spacer followed by a label or spacer followed by a solubilizing element or a PK enhancing element.

In another embodiment, the subject matter described herein pertains to a polypeptide comprising a sequence of Formula II (b):

Wherein: A is an electrophilic moiety, such as a Michael acceptor or a chloroethyl or bromoethyl group, which is capable of _reacting with a sulfhydryl group present on the residue X _{aa 13} to form a covalent thioether bond, thereby producing a macrocycle a peptide; wherein the thioether bond may or may not be oxidized to the corresponding diastereomeric hydrazide; and wherein A may be optionally present; and wherein if A is present, it may be Gly or other spacer having a free amine, _Cyclization of the peptide can be carried out by forming a _guanamine bond with a carboxyl group on the side chain of X _aa13 to obtain a cyclic peptide having an N-terminally linked to a _{guanamine in} the side chain; and wherein if A is absent, the X _aa1 residue The N-terminal amine group can be used for cyclization of the peptide via a _guanamine bond with a carboxyl group on the side chain of X _aa13 to obtain a cyclic peptide having an N-terminally linked to a _{guanamine in} the side chain; and wherein if A is present, It may be Gly or other spacer having a free amine end which may be used for cyclization of the peptide via a _{guanamine bond with} the C-terminal α-carboxyl group of X _aa14 to give an end-to-end cyclic peptide; If not present, the N-terminal amine group of the X _aa1 amino acid can be used to form a _{guanamine bond with} the C-terminal α-carboxyl group of X _{aa 14} Cyclization of the peptide to give an end-to-end cyclic peptide; X _aa1 is a natural or non-naturally occurring amino acid comprising Phe and Ala; X _aa2 is a natural or non-naturally occurring amino acid comprising ^m Phe and ^m Ala; X _aa3 is a natural or non-naturally occurring amino acid comprising ^m Nle and ^m Ala; X _aa4 is a natural or non-naturally occurring amino acid comprising ^m Gly, Gly and ^m Ala; X _aa5 is A naturally occurring or non-naturally occurring amino acid comprising Asp and Ala; X _aa6 is a natural or non-naturally occurring amino acid comprising Val (preferably) and Ala; X _aa7 is a naturally or non-naturally occurring amine group An acid comprising ^m Phe and Phe; X _aa8 is a natural or non-naturally occurring amino acid selected from the group consisting of Tyr and Ala; X _aa9 is a natural or non-naturally occurring amino acid comprising ^m Gly, ^m Ala and Gly; X _aa11 is a natural or non-naturally occurring amino acid comprising Tyr and Ala; X _aa12 is a natural or non-naturally occurring amino acid comprising Leu and Ala; X _aa13 is natural or unnatural the presence of amino acids comprising L-Cys, D-Cys, Asp, Glu and Gly; X _aa14 is Gly; or Gly, followed comprising at least two PEG diol units spacer; or Gly, followed comprising at least two ethylene glycol units PEG spacer, followed marked, such as biotin; wherein X _aa14 optionally present and wherein the amino acids of the C a terminal carbonyl carbon is attached to the nitrogen to form a carboxamide (NH ₂ ), an alkyl carboxamide (NHR ₁ ) or a dialkyl carboxamide (NR ₁ R ₂ ); wherein each of R ₁ and R ₂ is an alkyl group or An arylalkyl group; wherein if X _aa14 is absent, the C-terminal carbonyl carbon of X _{aa 13} is bonded to nitrogen to form carboxamide (NH ₂ ), alkylcarboxamide (NHR ₁ ) or dialkyl carboxamide ( NR ₁ R ₂ ); wherein R ₁ and R _{2 are} each an alkyl group or an arylalkyl group.

In another embodiment, the subject matter described herein pertains to a polypeptide comprising a sequence of Formula III (a):

Wherein: A is an organic or peptide linker between X _aa1 and X _aa12 to give a macrocyclic peptide; X _aa1 is a natural or non-naturally occurring aromatic or heteroaromatic or alkyl or heteroarylalkylamine group Acid; X _aa2 is a natural or non-naturally occurring alkyl or aromatic or charged amino acid; X _aa3 is a natural or non-naturally occurring aromatic or heteroaromatic or alkyl or heteroarylalkylamine Acid; X _aa4 is a natural or non-naturally occurring aromatic or heteroaromatic or alkyl or heteroarylalkylamino acid; X _aa5 is a natural or non-naturally occurring alkyl or heteroalkyl or aromatic or hetero An aromatic or heteroarylalkylamino acid; X _aa6 is a heteroaromatic or positively charged amino acid _which is naturally or non-naturally occurring; X _aa7 is a polar or charged amino acid which is naturally or non-naturally occurring. X _aa8 is a naturally or non-naturally occurring positively charged amino acid; X _aa9 is a natural or non-naturally occurring alkyl or heteroalkyl or aromatic or heteroaromatic or heteroarylalkylamino acid; _X aa10 is a naturally or non-naturally occurring, or of aromatic or heteroaromatic arylalkyl or heteroarylalkyl or heteroalkyl, or alkyl amino acid; X _aa11 is a naturally or non-naturally occurring The aromatic or heteroaromatic or arylalkyl or heteroarylalkyl or heteroalkyl or positively charged amino acid; X _aa12 is a naturally or non-naturally occurring amino acids of which may be suitably activated with a functional group reactive with one end of linker A to produce a cyclic peptide; X _aa13 is a naturally occurring or non-naturally occurring amino acid or spacer followed by a label or spacer followed by a solubilizing element or a PK enhancing element .

In another embodiment, the subject matter described herein pertains to a polypeptide comprising a sequence of Formula III (b):

Wherein: A is an electrophilic moiety, such as a Michael acceptor or a chloroethyl or bromoethyl group, which is capable of reacting with a sulfhydryl group of Cys ¹² to form a covalent thioether bond, thereby producing a macrocyclic peptide; wherein the sulfur The ether bond may or may not be oxidized to the corresponding diastereomeric hydrazide; X _aa1 is selected from Phe and D-Phe; X _aa2 is selected from Leu, Arg and Phe; X _aa3 is selected from Ile, Leu and Phe; X _aa4 is selected from Val, Tyr and Phe; X _aa5 is selected from Ile and Val; X _aa6 is selected from Arg and His; X _aa8 is selected from Arg; X _aa9 is selected from Val, Leu, Tyr and Phe; _Aa11 is selected from Arg and Tyr; X _aa13 is Gly; or Gly, followed by a PEG spacer comprising at least two ethylene glycol units, wherein X _{aa13 is} optionally present and wherein the C-terminal carbonyl carbon of the amino acid is linked To nitrogen to form carboxamide (NH ₂ ), alkyl carboxamide (NHR ₁ ) or dialkyl carboxamide (NR ₁ R ₂ ); wherein R ₁ and R _{2 are} each alkyl or arylalkyl Wherein if X _aa13 is absent, the C-terminal carbonyl carbon of Cys ¹² is attached to the nitrogen to form carboxamide (NH ₂ ), alkyl carboxamide (NHR ₁ ) or dialkyl carboxamide (NR ₁ R ₂ Wherein R ₁ and R _{2 are} each an alkyl or arylalkyl group.

In another embodiment, the subject matter described herein pertains to a polypeptide comprising a sequence of Formula I(d):

Wherein: A is a chloroethyl group attached to the α-amine of the N-terminal X _aa1 residue, and the chloroethyl group can react with the _sulfhydryl group present on the residue X _{aa 14} to form a covalent thioether bond, thereby Obtaining a macrocyclic peptide; wherein the thioether bond may or may not be oxidized to the corresponding diastereomeric hydrazide; and wherein X _aa1 is selected from the group consisting of L-Phe, L-Trp, L-Tyr, L-Phe(4-OMe), L-Phe(4-F), L-Phe(4-Cl), L-Phe(4-Br), L-Phe(4-Me), L-Phe(4 -CF ₃ ), L-1-Nal, L-2-Nal, L-Bip, L-3-Pya, L-4-Pya, L-3-Tha; X _aa2 is selected from L-Ala, L- ^a group consisting of ^m Ala and ^m Gly; X _aa3 is selected from the group consisting of L-Ala and L-Asn; X _aa4 is selected from the group consisting of L-Pro, L-Ala, L-α-Me-Pro , L-Pro (4-OH), L-Pro (4-NH ₂ ), L-Pro (4S-Ph), L-Azt, L-Pip and L-Oic; X _aa5 is selected from L-Ala, a group consisting of L-His and L-Leu; X _aa6 is selected from the group consisting of L-Ala, L-Arg, L-Asp, L-His and L-Leu; X _aa7 is selected from the group consisting of : ^m Gly, Gly, L- ^m Ala, D- ^m Ala, L-Pro, L-Ser, L- ^m Ser, L-Dab, L-Arg and L-His; X _aa9 is selected from L-Ala, LA a group consisting of rg and L-Ser; X _aa10 is selected from the group consisting of L-Trp, L-Met and L-Bzt; X _aa11 is selected from the group consisting of L-Nle, L- ^m Nle, L- ^m Ala, L-Phe, L- ^m Phe and L- ^m Leu and L- ^m Ser; X _aa12 is selected from the group consisting of L- ^m Nle and L- ^m Ala; X _aa13 is selected from L-Ala, L a group consisting of -Arg and L-Leu; X _aa14 is selected from the group consisting of L-Cys and D-Cys; X _aa15 is Gly; or Gly, followed by a PEG spacer comprising twelve ethylene glycol units; Wherein X _{aa15 is} optionally present and wherein the C-terminal carbonyl carbon of the amino acid is attached to the nitrogen to form _{carboxyguanamine} (CONH ₂ ); wherein if X _{aa 15} is absent, the C-terminal carbonyl carbon of X _{aa 14} is attached to the nitrogen Carboxyguanamine (CONH ₂ ) is formed.

In another embodiment, the subject matter described herein pertains to a polypeptide comprising a sequence of Formula II (c):

Wherein: A is a chloroethyl group attached to the α-amine of the N-terminal X _aa1 residue, and the chloroethyl group can react with a sulfhydryl group present on the Cys ¹³ residue to form a covalent thioether bond, thereby Obtaining a macrocyclic peptide; wherein the thioether bond may or may not be oxidized to the corresponding diastereomeric hydrazide; and wherein X _aa2 is selected from the group consisting of L- ^m Ala and L- ^m Phe; X _aa3 is selected from a group consisting of L- ^m Ala and L- ^m Nle; X _aa4 is selected from the group consisting of Gly, ^m Gly and L- ^m Ala; X _aa5 is selected from the group consisting of L-Ala and L-Asp; X _aa6 Selecting a group consisting of L-Ala and L-Val; X _aa7 is selected from the group consisting of L-Phe and L- ^m Phe; X _aa8 is selected from the group consisting of L-Ala and L-Tyr; X _aa9 is selected a group consisting of free Gly, ^m Gly and L- ^m Ala; X _aa12 is selected from the group consisting of L-Leu and L-Ala; X _aa14 is Gly; or Gly, followed by twelve ethylene glycol units PEG spacer; wherein X _aa14 carbonyl carbon of the C-terminal, or C-terminal carbonyl carbon of X _aa14, followed by a PEG spacer attached to the nitrogen to form a sub-2carboxamide (CONH _2).

In another embodiment, the subject matter described herein pertains to a polypeptide comprising a sequence of Formula III (c):

Wherein: A is a chloroethyl group attached to the α-amine of the N-terminal L-Phe residue, and the chloroethyl group can react with the sulfhydryl group present on the L-Cys ¹² residue to form a covalent thioether. a bond to produce a macrocyclic peptide; and wherein X _aa2 is selected from the group consisting of L-Leu, L-Arg, and L-Phe; X _aa3 is selected from the group consisting of L-Ile and L-Phe; and X _aa4 is selected from L -Phe, L-Tyr and L-Val; X _aa5 is selected from the group consisting of L-Ile and L-Val; X _aa9 is selected from the group consisting of L-Leu, L-Phe, L-Tyr and L-Val; wherein Cly ¹³ The terminal carbonyl carbon is attached to the nitrogen to form carboxamide (CONH ₂ ).

The invention also relates to macrocyclic peptides comprising the sequences provided by Formula I.

The invention also relates to macrocyclic peptides comprising the sequence provided by formula I(a).

The invention also relates to macrocyclic peptides comprising the sequences provided by formula I(b).

The invention also relates to macrocyclic peptides comprising the sequences provided by formula I(c).

The invention also relates to macrocyclic peptides comprising the sequence provided by formula I(d).

The invention also relates to macrocyclic peptides comprising the sequences provided by Formula II.

The invention also relates to macrocyclic peptides comprising the sequence provided by formula II (a).

The invention also relates to macrocyclic peptides comprising the sequence provided by formula II (b).

The invention also relates to macrocyclic peptides comprising the sequence provided by formula II (c).

The invention also relates to macrocyclic peptides comprising the sequence provided by Formula III.

The invention also relates to macrocyclic peptides comprising the sequence provided by formula III(a).

The invention also relates to macrocyclic peptides comprising the sequence of formula III (b).

The invention also relates to macrocyclic peptides comprising the sequence of formula III (c).

The invention also relates to macrocyclic peptides comprising the sequences provided by Formula IV.

The invention also relates to macrocyclic peptides comprising the sequence provided by Formula V.

The invention also relates to macrocyclic peptides comprising the sequences provided by Formula VI.

The invention also relates to macrocyclic peptides comprising the sequence provided by formula VII.

The present invention also relates to a macrocyclic peptide comprising a sequence selected from the group consisting of Compound No. 1, No. 2, No. 3, No. 4, No. 71, and No. 99.

The invention also relates to macrocyclic peptides comprising a sequence selected from the peptides described herein.

The invention also relates to methods of using the macrocyclic peptides of the invention to ameliorate and/or treat proliferative disorders and/or viral disorders.

The invention also relates to a method of modulating an immune response in an individual comprising administering to the individual one or more macrocyclic peptides comprising a sequence selected from the peptides described herein.

The invention also relates to a method of enhancing, stimulating or enhancing an immune response in an individual comprising administering to the individual one or more macrocyclic peptides comprising a sequence selected from the peptides described herein.

The invention also relates to a method of promoting inhibition of tumor cell growth by an immune system in an individual comprising administering to the individual a therapeutically effective amount of one or more macrocyclic peptides comprising a sequence selected from the peptides described herein.

The invention also relates to a method of treating an infectious disease in an individual comprising administering to the individual a therapeutically effective amount of one or more macrocyclic peptides comprising a sequence selected from the peptides described herein.

The invention also relates to a combination comprising a sequence selected from the macrocyclic peptides described herein, and An agent, such as an antimicrobial therapy, an antiviral therapy, another immunomodulatory therapy, a vaccine, or a cancer chemotherapeutic agent.

According to the present invention, peptides which specifically bind to PD-L1 and are capable of inhibiting the interaction of PD-L1 with PD-1 and CD80 have been found. These macrocyclic peptides exhibit in vitro immunomodulatory effects, making them therapeutic candidates for the treatment of various diseases including cancer and infectious diseases.

The term "specifically binds" refers to the interaction between a protein and a binding molecule, such as a compound or ligand. The interaction is related to the presence of a specific structure (ie, an enzyme binding site, an antigenic determinant or an epitope) of the protein recognized by the binding molecule. For example, if a compound specifically binds to a protein binding site "A", the compound will be present in a reaction containing a protein comprising binding site A and a labeled peptide that specifically binds to protein binding site A. The amount of labeled peptide to protein is reduced. In contrast, non-specific binding of a compound to a protein does not result in the labeled peptide leaving the protein due to concentration-dependent substitution.

Other embodiments include polypeptides comprising the following structures: or or or

Another embodiment is a pharmaceutical composition comprising Formulas I(a), I(b), I(c), II(a), II(b), III(a), III(b), IV, V A polypeptide of VI, VII, or a peptide comprising at least one macrocyclic peptide described herein.

Another embodiment relates to a pharmaceutical combination comprising Formulas I(a), I(b), I(c), II(a), II(b), III(a), III(b), IV, V a polypeptide of VI, VII or VII or a macrocyclic peptide as described herein, and at least one therapeutic agent selected from the group consisting of an antimicrobial agent, an antiviral agent, an anticancer agent, an antidiabetic agent, an antiobesity agent, an antihypertensive agent Agent, anti-atherosclerosis agent and lipid-lowering agent.

Another embodiment relates to Formula I(a), I(b), I(c), II(a), II(b), III(a), III(b), IV, V, VI or VII A pharmaceutical combination of a polypeptide or a macrocyclic peptide described herein with other agents disclosed herein.

Another embodiment relates to a method of treating or delaying the progression or onset of a cancer and/or virological condition comprising administering to a mammalian species in need of treatment a therapeutically effective amount of Formula I(a), I(b), I ( c), a polypeptide of II (a), II (b), III (a), III (b), IV, V, VI or VII, or a macrocyclic peptide as described herein.

The invention is intended to include all isotopes of the atoms present in the compounds of the invention. Isotopes include those atoms having the same atomic number but different mass numbers. By way of example and not limitation, hydrogen isotopes include deuterium and tritium. Carbon isotopes include ¹³ C and ¹⁴ C. Isotopically labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art or by methods similar to those described herein, using isotopically labeled suitable reagents in place of other non-labeling reagents used. . These compounds can have a variety of potential uses, for example, as standards and reagents for determining biological activity. In the case of stable isotopes, such compounds may have the potential to advantageously modify biological, pharmacological or pharmacokinetic properties.

Another aspect of the subject matter described herein is the disclosed peptide as a radiolabeled ligand for the development of ligand binding assays or for monitoring in vivo adsorption, metabolism, distribution, receptor binding or occupancy, or compound disposal. use. For example, the macrocyclic peptides described herein can be prepared using the radioisotope ¹²⁵ I and the resulting radiolabeled peptides can be used to develop binding assays or for metabolic studies. Alternatively, and for the same purpose, the macrocyclic peptides described herein can be converted to the radiolabeled form by catalytic deuteration using methods known to those skilled in the art.

The macrocyclic peptides of the present invention can also be used as PET imaging agents by the addition of radiotracers by methods known to those skilled in the art.

Preferred peptides include at least one macrocyclic peptide provided herein and such peptides can be included in pharmaceutical compositions and combinations.

The definitions provided herein, without limitation, apply to the terms used throughout the specification, unless otherwise limited in the particular circumstances.

It is known to those skilled in the art of amino acids and peptides that amino acids include compounds represented by the following general structure:

Wherein R and R' are as discussed herein.

Unless otherwise stated, the term "amino acid" as used herein alone or as part of another group includes, without limitation, an amine group and a carboxyl group attached to the same carbon (referred to as "alpha" carbon), wherein R And/or R' may be a natural or non-natural side chain, including hydrogen. The absolute "S" configuration at the "α" carbon is often referred to as the "L" or "natural" configuration. In the case where both "R" and "R" (the small oxime in the upper right corner of the letter) are hydrogen, the amino acid is glycine and does not have a palm.

As used herein, the term "naturally occurring amino acid side chain" refers to any naturally occurring amino acid (ie, alanine, arginine, aspartic acid, aspartic acid, cysteine, Gluten, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, valine, serine, threonine, The side chain of tryptophan, tyrosine and valine acid, which is usually in the S configuration (ie, L-amino acid).

As used herein, the term "non-naturally occurring amino acid side chain" refers to the side chain of any naturally occurring amino acid (ie, D-amino acid), typically in the R configuration, or a group other than the naturally occurring amino acid side chain (ie, D- or L-amino acid, respectively) of the S configuration selected from: C ₂ -C ₇ alkenyl, C ₁ -C ₃ alkane Oxy C ₁ -C ₃ alkyl, C ₁ -C ₆ alkoxycarbonyl C ₁ -C ₃ alkyl, C ₁ -C ₇ alkyl, C ₁ -C ₃ alkylthio C ₁ -C ₃ alkyl , amidino C ₁ -C ₃ alkyl, amino C ₁ -C ₃ alkyl, azaindenyl C ₁ -C ₃ alkyl, benzothiazolyl C ₁ -C ₃ alkyl, benzothiophene C ₁ -C ₃ alkyl, benzyloxy C ₁ -C ₃ alkyl, carboxy C ₁ -C ₃ alkyl, C ₃ -C ₆ cycloalkyl C ₁ -C ₃ alkyl, diphenyl , furyl C ₁ -C ₃ alkyl, imidazolyl C ₁ -C ₃ alkyl, naphthyl C ₁ -C ₃ alkyl, pyridyl C ₁ -C ₃ alkyl, thiazolyl C ₁ -C ₃ alkane a thiophenyl-C ₁ -C ₃ alkyl group; a biphenyl C ₁ -C ₃ alkyl group, wherein the biphenyl is optionally substituted by a methyl group; a mercapto C ₁ -C ₃ alkyl group, wherein the thiol moiety is optionally One selected from C ₁ -C ₃ alkyl, carboxy C ₁ -C ₃ alkyl, halo, hydroxy And a phenyl group substitution wherein the phenyl group is further substituted with one, two or three groups independently selected from the group consisting of C ₁ -C ₃ alkoxy, C ₁ -C ₃ alkyl and halo; NR ^a R ^b (C ₁ -C ₇ alkyl), wherein R ^a and R ^{b are} independently selected from hydrogen, C ₂ -C ₄ alkenyloxycarbonyl, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkylcarbonyl, C ₃ -C ₆ cycloalkylcarbonyl, furylcarbonyl and phenylcarbonyl. When the alkyl linker contains more than one carbon, another NR ^a R ^b group may be present on the chain.

NR ^c R ^d carbonyl C ₁ -C ₃ alkyl, wherein R ^c and R ^{d are} independently selected from hydrogen, C ₁ -C ₃ alkyl and triphenylmethyl; phenyl C ₁ -C ₃ alkyl, wherein benzene The base moiety is optionally substituted with one, two, three, four or five groups independently selected from C ₁ -C ₄ alkoxy, C ₁ -C ₄ alkyl, C ₁ -C ₃ alkylsulfonium Amino, amidino, amine, amine C ₁ -C ₃ alkyl, aminosulfonyl, carboxyl, cyano, halo, halo C ₁ -C ₃ alkyl, hydroxy, -NC ( NH ₂ ) ₂ , nitro and -OP(O)(OH) ₂ ; and phenoxy C ₁ -C ₃ alkyl, wherein the phenyl group is optionally substituted by a C ₁ -C ₃ alkyl group.

As used herein, the term "C ₂ -C ₄ alkenyl group" means containing at least one carbon-carbon double bond, a linear or branched chain group of two to four carbon atoms.

As used herein, the term "C ₂ -C ₇ alkenyl group" means containing at least one carbon-carbon double bond, a linear or branched chain group of two to seven carbon atoms.

As used herein, the term "C ₂ -C ₄ alkenyl group" means connected to the parent molecular moiety of C ₂ -C ₄ alkenyl group via an oxygen atom.

As used herein, the term "C ₁ -C ₃ alkoxy" means C is connected to the parent molecular moiety ₁ -C ₃ alkyl via an oxygen atom.

As used herein, the term "C ₁ -C ₄ alkoxy" refers to the connection to the parent molecular moiety C ₁ -C ₄ alkyl group via an oxygen atom.

As used herein, the term "C ₁ -C ₆ alkoxy" means C is connected to the parent molecular moiety ₁ -C ₆ alkyl via an oxygen atom.

As used herein, the term "C ₁ -C ₃ alkoxy C ₁ -C ₃ alkyl" refers to a C ₁ -C ₃ alkoxy group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group.

As used herein, the term "C ₁ -C ₆ alkoxycarbonyl" refers to a C ₁ -C ₆ alkoxy group attached to the parent molecular moiety through a carbonyl group.

As used herein, the term "C ₁ -C ₆ alkoxycarbonyl C ₁ -C ₃ alkyl" refers to a C ₁ -C ₆ alkoxycarbonyl group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group.

As used herein, the term "C ₁ -C ₃ alkyl" refers to a radical derived from a straight or branched chain saturated hydrocarbon containing one to three carbon atoms.

As used herein, the term "C ₁ -C ₄ alkyl" refers to a radical derived from a straight or branched chain saturated hydrocarbon containing from one to four carbon atoms.

As used herein, the term "C ₁ -C ₆ alkyl" refers to a radical derived from a straight or branched chain saturated hydrocarbon containing from one to six carbon atoms.

As used herein, the term "C ₁ -C ₃ alkylcarbonyl" refers to a C ₁ -C ₃ alkyl group attached to the parent molecular moiety through a carbonyl group.

As used herein, the term "C ₁ -C ₃ alkylthio atom" refers to a C ₁ -C ₃ alkyl group attached to the parent molecular moiety through a sulfur atom.

As used herein, the term "C ₁ -C ₃ alkylthio C ₁ -C ₃ alkyl" refers to a C ₁ -C ₃ alkylthio group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group.

As used herein, the term "C ₁ -C ₃ alkylsulfonyl" refers to a C ₁ -C ₃ alkylthio group attached to the parent molecular moiety through a sulfonyl group.

As used herein, the term "C ₁ -C ₃ alkylsulfonylamino" refers to a C ₁ -C ₃ alkylsulfonyl group attached to the parent molecular moiety via an amine group.

As used herein, the term "acyl group" means -C (O) NH _2.

As used herein, the term "acyl C ₁ -C ₃ alkyl group" means acyl group connected to the parent molecular moiety via a C ₁ -C ₃ alkyl.

As used herein, the term "amino" means -NH _2.

As used herein, the term "amino C ₁ -C ₃ alkyl" refers to an amine group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group.

As used herein, the term "aminosulfonyl" refers to an amine group attached to the parent molecular moiety through a sulfonyl group.

As used herein, the term "aza-indolyl C ₁ -C ₃ alkyl" means azaindole connected to the parent molecular group via a C ₁ -C ₃ alkyl. Azaindolyl can be attached to the alkyl moiety via any substitutable atom in the group.

As used herein, the term "benzothiazolyl C ₁ -C ₃ alkyl" refers to a benzothiazolyl group attached to the parent molecule via a C ₁ -C ₃ alkyl group. The benzothiazolyl group can be attached to the alkyl moiety via any substitutable atom in the group.

As used herein, the term "benzothienyl C ₁ -C ₃ alkyl" refers to a benzothienyl group attached to the parent molecule via a C ₁ -C ₃ alkyl group. The benzothienyl group can be attached to the alkyl moiety via any substitutable atom in the group.

As used herein, the term "benzyloxy" refers to a benzyl group attached to the parent molecular moiety through an oxygen atom.

As used herein, the term "benzyloxy-C ₁ -C ₃ alkyl" means benzyloxy connected to the parent molecular moiety via a C ₁ -C ₃ alkyl.

As used herein, the term "biphenyl C ₁ -C ₃ alkyl" refers to a biphenyl group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group. The biphenyl group can be attached to the alkyl moiety via any substitutable atom in the group.

As used herein, the term "carbonyl" refers to -C(O)-.

As used herein, the term "carboxy" refers to -CO ₂ H.

As used herein, the term "carboxy C ₁ -C ₃ alkyl" refers to a carboxy group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group.

As used herein, the term "cyano" refers to -CN.

As used herein, the term "C ₃ -C ₆ cycloalkyl" means a saturated monocyclic hydrocarbon ring system having three to six carbon atoms and zero heteroatoms.

As used herein, the term "C ₃ -C ₆ cycloalkyl C ₁ -C ₃ alkyl" refers to a C ₃ -C ₆ cycloalkyl group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group.

As used herein, the term "C ₃ -C ₆ cycloalkylcarbonyl" refers to a C ₃ -C ₆ cycloalkyl group attached to the parent molecular moiety through a carbonyl group.

As used herein, the term "furyl C ₁ -C ₃ alkyl" refers to a furanyl group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group. A furyl group can be attached to the alkyl moiety via any substitutable atom in the group.

As used herein, the term "furanylcarbonyl" refers to a furanyl group attached to the parent molecular moiety through a carbonyl group.

As used herein, the terms "halo" and "halogen" mean F, Cl, Br or I.

As used herein, the term "halo C ₁ -C ₃ alkyl" refers to a C ₁ -C ₃ alkyl group substituted with one, two or three halogen atoms.

As used herein, the term "halomethyl" refers to a methyl group substituted with one, two or three halogen atoms.

As used herein, the term "hydroxy" refers to -OH.

As used herein, the term "imidazolyl C ₁ -C ₃ alkyl" refers to an imidazolyl group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group. The imidazolyl group can be attached to the alkyl moiety via any substitutable atom in the group.

As used herein, the term "mercapto C ₁ -C ₃ alkyl" refers to a fluorenyl group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group. A mercapto group can be attached to the alkyl moiety via any substitutable atom in the group.

As used herein, the term "naphthyl C ₁ -C ₃ alkyl" refers to a naphthyl group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group. A naphthyl group can be attached to the alkyl moiety via any substitutable atom in the group.

As used herein, the term "nitro" refers to -NO ₂ .

As used herein, the term "NR ^a R ^b " refers to two groups R ^a and R ^b attached to the parent molecular moiety through a nitrogen atom. R ^a and R ^{b are} independently selected from the group consisting of hydrogen, C ₂ -C ₄ alkenyloxycarbonyl, C ₁ -C ₃ alkylcarbonyl, C ₃ -C ₆ cycloalkylcarbonyl, furylcarbonyl and phenylcarbonyl.

As used herein, the term "NR ^a R ^b (C ₁ -C ₃ )alkyl" refers to a NR ^a R ^b group attached to the parent molecular moiety via a C ₁ -C ₃ alkyl group.

As used herein, the term "NR ^c R ^d " refers to two groups R ^c and R ^d attached to the parent molecular moiety through a nitrogen atom. R ^c and R ^{d are} independently selected from the group consisting of hydrogen, C ₁ -C ₃ alkyl and triphenylmethyl.

As used herein, the term "NR ^c R ^d carbonyl" refers to an NR ^c R ^d group attached to the parent molecular moiety through a carbonyl group.

As used herein, the term "NR ^c R ^d carbonyl C ₁ -C ₃ alkyl" refers to an NR ^c R ^d carbonyl group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group.

As used herein, the term "phenoxy" refers to a phenyl group attached to the parent molecular moiety through an oxygen atom.

As used herein, the term "phenoxy C ₁ -C ₃ alkyl" refers to a phenoxy group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group.

As used herein, the term "phenyl C ₁ -C ₃ alkyl" refers to a phenyl group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group.

As used herein, the term "phenylcarbonyl" refers to a phenyl group attached to the parent molecular moiety through a carbonyl group.

As used herein, the term "pyridyl C ₁ -C ₃ alkyl" refers to a pyridyl group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group. A pyridyl group can be attached to the alkyl moiety via any substitutable atom in the group.

As used herein, the term "thio" refers to -S-.

As used herein, the term "sulfonic acyl" means -SO ₂ -.

As used herein, the term "thiazolyl C ₁ -C ₃ alkyl" refers to a thiazolyl group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group. The thiazolyl group can be attached to the alkyl moiety via any substitutable atom in the group.

As used herein, the term "thienyl C ₁ -C ₃ alkyl" refers to a thienyl group attached to the parent molecular moiety through a C ₁ -C ₃ alkyl group. A thienyl group can be attached to the alkyl moiety via any substitutable atom in the group.

The term "treatment" means: (i) preventing a disease, disorder or condition in a patient who is susceptible to an infectious disease, disorder and/or condition, but has not yet been diagnosed; (ii) inhibiting the disease, disorder or condition, ie And (iii) mitigating the disease, condition or condition, that is, causing the disease, condition and/or condition and/or symptoms associated with the disease, condition and/or condition to resolve.

The term "alkyl" as used herein, alone or as part of another group, unless otherwise indicated, includes, without limitation, from 1 to 40 carbons, preferably from 1 to 20 carbons, more preferably from 1 to about 3 in the normal chain. Linear and branched hydrocarbons of 8 carbons, such as methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4, 4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, decyl, decyl, undecyl, dodecyl, various branched isomers thereof, and the like Things. Furthermore, an alkyl group, as defined herein, may be optionally substituted on any available carbon atom via one or more functional groups typically attached to the chain, such as, but not limited to, alkyl, aryl, alkenyl, alkynyl, Hydroxy, arylalkyl, cycloalkyl, cycloalkylalkyl, alkoxy, arylalkoxy, heteroaryloxy, heteroarylalkoxy, alkanoyl, halo, hydroxy, thio , nitro, cyano, carboxyl, carbonyl ( Carboxylamido group, amine group, alkylamino group, dialkylamino group, decylamino group, alkylamino group, aryl decylamino group, heteroaryl guanylamino group, azide group, fluorenyl group, Mercapto, phosphonate, phosphinate, sulfonate, sulfonylamino, haloaryl, CF ₃ , OCF ₂ , OCF ₃ , aryloxy, heteroaryl, cycloalkylalkoxyalkyl a cycloheteroalkyl group and the like to form an alkyl group such as trifluoromethyl, 3-hydroxyhexyl, 2-carboxypropyl, 2-fluoroethyl, carboxymethyl, cyanobutyl and the like .

Unless otherwise stated, the term "cycloalkyl" as used herein alone or as part of another group includes, without limitation, saturated or partially unsaturated (containing) one to three attached or fused rings (including 1 or 2 double bond) cyclic hydrocarbon groups, including monocyclic alkyl groups, bicycloalkyl groups and tricycloalkyl groups condensable with one aromatic ring as described for aryl groups, which contain a total of 3 to 20 Forming a ring of carbon, preferably 4 to 7 carbons to form each ring; including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, cyclododecyl Cyclohexenyl, Any of such groups may optionally be substituted on any available carbon atom with one or more groups selected from the group consisting of hydrogen, halo, haloalkyl, alkyl, haloalkyl, alkoxy, Haloalkoxy, alkenyl, trifluoromethyl, trifluoromethoxy, alkynyl, cycloalkylalkyl, decyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, heteroaryl, Arylalkyl, aryloxy, aryloxyalkyl, arylalkoxy, arylthio, arylazo, heteroarylalkyl, heteroarylalkenyl, heteroarylheteroaryl, Heteroaryloxy, hydroxy, nitro, pendant oxy, cyano, carboxy, carbonyl ( Carboxylammonium, amine, substituted amine (wherein the amine group includes 1 or 2 substituents which are alkyl, aryl or other aryl compounds mentioned in the definition) , amidino, azide, fluorenyl, methionyl, phosphonic acid, hypophosphorous, sulfonate, sulfonylamino, thiol, alkylthio, arylthio, heteroarylthio, Arylthioalkyl, alkoxyarylthio, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, arylaminocarbonyl, alkoxycarbonyl, aminocarbonyl, alkylcarbonyloxy, arylcarbonyl An oxy group, an alkylcarbonylamino group, an arylcarbonylamino group, an arylsulfinyl group, an arylsulfinylalkyl group, an arylsulfonylamino group or an arylsulfonylaminocarbonyl group, or Any of the alkyl substituents stated.

The term "aryl" as used herein, alone or as part of another group, refers to, without limitation, monocyclic and bicyclic aromatic groups (such as phenyl or naphthyl) having from 6 to 10 carbons in the ring portion. And optionally including one to three other rings fused to an "aryl" group (such as an aryl, cycloalkyl, heteroaryl or heterocycloalkyl ring) and optionally on any available carbon atom. Substituted with one or more groups selected from the group consisting of hydrogen, alkyl, halo, haloalkyl, alkoxy, haloalkoxy, alkenyl, trifluoromethyl, trifluoromethoxy, alkynyl, Cycloalkylalkyl, fluorenyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, heteroaryl, arylalkyl, aryloxy, aryloxyalkyl, arylalkoxy, aryl Thio, aryl azo, heteroarylalkyl, heteroarylalkenyl, heteroaryloxy, heteroarylalkoxy, heteroarylalkoxyalkyl, hydroxy, nitro, pendant oxy a cyano group, an amine group, a substituted amine group (wherein the amine group includes 1 or 2 substituents which are an alkyl group, a cycloalkyl group, a heterocycloalkyl group, a heteroaryl group or an aryl group or are mentioned in the definition Any of the other aryl compounds)), thiol, alkylthio, arylthio, heteroarylthio, arylthioalkyl, alkoxyarylthio, alkylcarbonyl, arylcarbonyl, alkane Aminocarbonyl, cycloalkylaminocarbonyl, arylaminocarbonyl, heteroarylaminocarbonyl, heteroarylalkylaminocarbonyloxycarbonyl, aminocarbonyl, alkylcarbonyloxy, arylcarbonyl Oxyl, alkylcarbonylamino, arylcarbonylamino, Alkylsulfinyl acyl, sulfo arylalkylene acyl group, an aryl group or an aryl group sulfo acyl group Sulfonic aminocarbonyl, or any one of the above-stated alkyl substituents.

The term "arylalkyl" as used herein, alone or as part of another group, refers to, without limitation, an alkyl group, as defined above, having an aryl substituent, such as benzyl, phenethyl or naphthalene. A propyl group, wherein the aryl group and/or alkyl group may be substituted as defined above.

"Alkoxy", "aryloxy", "heteroaryloxy", "arylalkoxy" or "heteroarylalkoxy" as used herein, alone or as part of another group, includes (not limited to) an alkyl or aryl group as defined above attached via an oxygen atom.

As used herein, the term "heterocycle" or "heterocyclyl" means, without limitation, an unsubstituted or substituted stable 4 member, 5 member, 6 member or 7 membered single ring system which may be saturated or unsaturated. And consisting of carbon atoms and one to four heteroatoms selected from the group consisting of nitrogen, sulfur, oxygen and/or SO or SO ₂ groups, wherein the nitrogen and sulfur heteroatoms may be oxidized as appropriate, and the nitrogen heteroatoms may optionally be It is quaternized. The heterocycle can be attached at any heteroatom or carbon atom which results in a stable structure. Examples of such heterocyclic groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, piperazinyl, pendant oxypyrrolidinyl, pendant oxypiperazinyl, side oxygen Piperidinyl and oxadiazolyl. The heterocyclic group may be optionally substituted with one or more functional groups, such as those described for "alkyl" or "aryl".

The term "heterocycloalkyl" as used herein, alone or as part of another group, refers to, without limitation, an alkyl group as defined above having a heterocycloalkyl substituent, wherein the "heterocyclyl" / or alkyl may be substituted as defined above.

As used herein, the term "heteroaryl" refers to, without limitation, a 5-membered, 6-membered or 7-membered aromatic heterocyclic ring containing one or more selected from the group consisting of nitrogen, sulfur, oxygen, and/or SO or SO ₂ groups. The hetero atom of the group. Such rings may be fused to another aryl or heteroaryl ring and include possible N-oxides; examples of such heteroaryl groups include, but are not limited to, furan, pyrrole, thiophene, pyridine, pyrimidine, pyrazine , pyridazine, isoxazole, oxazole, imidazole and the like. The heteroaryl group may be optionally substituted with one or more functional groups which are typically attached to the chain, such as those described for "alkyl" or "aryl".

The term "heteroarylalkyl" as used herein, alone or as part of another group, refers to, without limitation, an alkyl group as defined above having a heteroaryl substituent wherein the heteroaryl group and/or The alkyl group may be substituted as defined above.

The "inhibitory concentration" of the PD-1/PD-L1 inhibitor means the concentration at which the interaction of PD-1 and PD-L1 inhibits the measurable percentage of the compound screened in the assay of the present invention. Examples of the range of values of the "inhibitory concentration" of IC ₅₀ to IC _90, and preferably _{IC 50, IC 60, IC 70} , IC 80 or IC _90, which represent the PD-1 / PD-L1 binding activity is decreased by 50% 60%, 70%, 80% or 90%. More preferably, the "inhibitory concentration" is measured by the IC ₅₀ value. It should be understood that another name for IC ₅₀ is the half maximum inhibitory concentration.

The binding of the macrocyclic peptide to PD-L1 can be measured by, for example, homogenous time difference fluorescence (HTRF), surface plasma resonance (SPR), isothermal titration calorimetry (ITC), nuclear magnetic resonance spectroscopy. Method (NMR) and the like. Furthermore, the binding of macrocyclic peptides to PD-L1 expressed on the cell surface can be measured in cell binding assays as described herein.

Administration of a therapeutic agent described herein includes, without limitation, administration of a therapeutically effective amount of a therapeutic agent. The term "therapeutically effective amount" as used herein refers to, without limitation, the amount of a therapeutic agent that treats or prevents a condition, which can be administered by administering a PD-1/PD-L1 binding inhibitor as described herein. The composition is treated. This amount is an amount sufficient to exhibit a detectable therapeutic or prophylactic or ameliorating effect. Such effects can include, for example and without limitation, treating or preventing the conditions listed herein. The exact and effective amount of an individual will depend on the individual's body and health, the nature and extent of the condition being treated, the advice of the treating physician, and the therapeutic or therapeutic combination selected for administration. Therefore, pre-specifying the exact effective amount does not apply.

The macrocyclic peptide of the present invention showed strong binding activity to PD-L1 in both HTRF analysis and cell binding assay. In addition, macrocyclic peptides also show biological activity in CMV recall and HIVElispot analysis shows that it can be used to ameliorate and/or treat proliferative disorders such as cancer, and virological indications, including HIV.

In another aspect, the invention relates to a method of inhibiting tumor cell growth in an individual using the macrocyclic peptides of the invention. As shown herein, the macrocyclic peptides of the invention are capable of binding to PD-L1, disrupting the interaction between PD-L1 and PD-1, and anti-PD-1 singles known to block interaction with PD-1 The antibody competes for binding to PD-L1, enhances the secretion of IFNγ by CMV-specific T cells, and enhances the secretion of IFNg by HIV-specific T cells. Thus, the macrocyclic peptides of the invention are useful for modulating an immune response, treating a disease (such as a cancer or an infectious disease), stimulating a protective autoimmune response, or stimulating an antigen-specific immune response (eg, by blocking the PD-L1 peptide) Co-administered with the antigen of interest).

In order to more easily understand the present invention, certain terms are first defined. Other definitions are set forth throughout the implementation.

The terms "stylized death ligand 1", "stylized cell death ligand 1", "protein PD-L1", "PD-L1", "PDL1", "PDCDL1", "hPD-L1", " hPD-LI", "CD274" and "B7-H1" are used interchangeably and include human PD-L1 variants, isoforms, homologous species, and at least one common antigen with PD-L1. An analog of a base. The complete PD-L1 sequence can be found under GENBANK® accession number NP_054862.

The terms "Stylized Death 1", "Stylized Cell Death 1", "Protein PD-1", "PD-1", "PD1", "PDCD1", "hPD-1" and "hPD-I" are interchangeable. It is used and includes human PD-1 variants, isoforms, homologous species, and analogs having at least one common epitope with PD-1. The complete PD-1 sequence can be found under GENBANK® accession number U64863.

The terms "cytotoxic T lymphocyte-associated antigen-4", "CTLA-4", "CTLA4", "CTLA-4 antigen" and "CD152" (see for example Murata, Am. J. Pathol. , 155: 453-460) (1999)) is used interchangeably and includes human CTLA-4 variants, isoforms, homologous species, and analogs having at least one common epitope with CTLA-4 (see, for example, Balzano, Int. J. Cancer Suppl. , 7:28-32 (1992)). The complete CTLA-4 nucleic acid sequence can be found under GENBANK® Accession No. L15006.

The term "immune response" refers to, for example, lymphocytes, antigen presenting cells, phagocytic cells, granulocytes, and the action of soluble macromolecules (including macrocyclic peptides, cytokines, and complements) produced by such cells or livers. Sexual pathogens, cells or tissues infected with pathogens, cancer cells or (in the case of autoimmune or pathological inflammation) normal human cells or tissues produce selective damage, destruction or exclusion from the human body.

"Signal transduction pathway" refers to the biochemical relationship between a variety of signal transduction molecules that function in transferring a signal from one part of a cell to another part of the cell. As used herein, the idiom "cell surface receptor" includes, for example, molecular and molecular complexes that are capable of receiving signals and that will conduct this signal through the plasma membrane of the cell. An example of the "cell surface receptor" of the present invention is the PD-1 receptor.

The term "macropeptide derivative" refers to any modified form of the macrocyclic peptide disclosed herein, such as a mutation, an isoform, a linker-modified peptide, an antibody, and/or another A combination of agents and the like.

As used herein, a macrocyclic peptide of the invention that "specifically binds to human PD-L1" means less than about 200 nM, less than about 150 nM, less than about 100 nM, less than about 80 nM, less than about 60 nM, less than about 40 nM, less than about 20 nM. , less than about 15nM, less than about 10 nM, less than about of 5 nM, less than about 1nM or less IC ₅₀ binding to a macrocyclic peptide of human PD-L1. In this context, the term "about" should be interpreted to mean greater or less than the stated amount ± 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, Any value between 15, 16, 17, 18, 19 or 20 nM.

The term "treatment" means the administration of an active agent for the purpose of treating, curing, alleviating, alleviating, altering, remedying, ameliorating, ameliorating or affecting a condition (eg, a disease), a symptom of a condition, or preventing or delaying symptoms, complications, or complications. The onset of a biochemical marker of the disease, or the arrest or inhibition of the development of a disease, condition or condition in a statistically significant manner.

As used herein, "adverse events" (AE) are any unfavorable and generally unintentional, even undesirable signs (including abnormal experimental findings), symptoms, or diseases associated with the use of medical treatment. For example, adverse events may be associated with activation of the immune system in response to treatment or expansion of immune system cells (eg, T cells) in response to treatment. The medical treatment can have one or more related AEs and each AE can have the same or a different severity. A reference to a method that can "change adverse events" means that the treatment regimen reduces the incidence and/or severity of one or more AEs associated with the use of different treatment regimens.

As used herein, "proliferative disease" refers to a condition in which cell growth is enhanced to exceed normal levels. For example, proliferative diseases or conditions include malignant diseases (eg, esophageal cancer, colon cancer, cholangiocarcinoma) and non-malignant diseases (eg, atherosclerosis, benign hyperplasia, and benign prostatic hypertrophy).

As used herein, "about" or "substantially encompasses" means that a particular value is within an acceptable tolerance as determined by those skilled in the art, depending in part on how the value is measured or determined, ie Limitations of the measurement system. For example, "about" or "basically" "Include" may mean within one or more standard deviations in accordance with the practice in the art. Alternatively, "about" or "substantially" may mean up to 20%. Moreover, particularly with respect to biological systems or methods, the term can mean up to an order of magnitude or at most 5 times. When a particular value is provided in the application and the scope of the patent application, the meaning of "about" or "substantially encompasses" is assumed to be within the acceptable tolerance for that particular value, unless otherwise stated.

Unless stated otherwise, any range of concentrations, percentage ranges, ratio ranges or integer ranges are to be understood to include any integer value within the range and, where appropriate, its fraction (such as one tenth of an integer) One hundredth of a percent).

competition analysis

The invention is also directed to being capable of at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, and at least about 100% The reference anti-PD-L1 antibody (MDX-1105) competes for binding to the macrocyclic peptide. Such macrocyclic peptides may share structural homology with one or more macrocyclic peptides disclosed herein (including mutants, conservative substitutions, functional substitutions and deletion forms, with the restriction that they specifically bind to PD-L1). For example, if the macrocyclic peptide and the reference anti-PD-L1 antibody bind substantially to the same region of PD-L1, the epitope of PD-L1 to which the macrocyclic peptide binds should be at least with the anti-PD-L1 monoclonal antibody. The combined PD-L1 epitope overlaps. The overlap region can range from one amino acid residue to several hundred amino acid residues. The macrocyclic peptide should then compete with the anti-PD-L1 monoclonal antibody and/or block the binding of the anti-PD-L1 monoclonal antibody to PD-L1, and thereby reduce the binding of anti-PD-L1 monoclonal antibody to PD-L1, in competition Preferably, the combination is reduced by at least about 50% in the analysis.

Anti-PD-L1 antibodies that can be used as reference antibodies for competitive assay purposes are known in the art. For example, the following representative anti-PD-L1 antibodies can be used: MDX-1105 (BMS); L01X-C (Serono), L1X3 (Serono), MSB-0010718C (Serono), and PD-L1 precursor antibody (CytomX) And the PD-L1 antibody disclosed in co-owned WO 2007/005874.

Anti-PD-1 antibodies that can be used as reference antibodies for competitive assay purposes are known in the art. For example, the following representative anti-PD-1 antibodies can be used: nivolumab (BMS); 17D8, 2D3, 4H1, 4A11, 7D3, and 5F4, each of which is disclosed in co-owned U.S. Patent No. 8,008,449 ( The antibody disclosed in U.S. Patent No. 7,488,802;

Variant macrocyclic peptide

In still another embodiment, the macrocyclic peptide of the invention comprises an amino acid sequence homologous to the amino acid sequence of the macrocyclic peptide described herein, and wherein the macrocyclic peptide retains the desired function of the macrocyclic peptide of the invention and/ Or biological characteristics.

For example, the invention provides a macrocyclic peptide or antigen binding portion thereof comprising: an amino acid sequence at least 80% homologous to an amino acid sequence selected from the compounds described herein; and exhibiting one or more of the following properties macrocyclic peptides: (a) macrocyclic peptides 200nM 200nM or less than the IC ₅₀ binding to human PD-L1; (b) a macrocyclic peptide does not substantially bind to human CD28, CTLA-4 or ICOS; (c) macrocycle Peptides enhance the secretion of IFNγ by CMV-specific T cells; (d) macrocyclic peptides enhance the secretion of IFNγ by HIV-specific T cells; (e) macrocyclic peptides bind to human PD-1 and one or more of the following: cynomolgus PD-1 , woodchuck PD-1 and/or mouse PD-1; (f) macrocyclic peptide inhibits PD-L1 and/or PD-L2 binding to PD-1; (g) macrocyclic peptide is capable of interacting with PD-1 Monoclonal antibodies (including nivozumab (BMS-936558, MDX-1106)) compete for binding; (h) macrocyclic peptides inhibit tumor cell growth in cell analysis and/or in vivo assays; and/or (i) large Cyclic peptides inhibit HIV in cellular assays and/or in vivo assays.

In other embodiments, the macrocyclic peptide amino acid sequence can be about the sequence listed above 80%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96% , about 97%, about 98% or about 99% homologous. In this context, the term "about" should be interpreted to mean any value between 1%, 2%, 3%, 4% or 5% greater or less than the stated amount. A macrocyclic peptide of the invention having a sequence that is highly consistent with the sequences listed above (i.e., 80% or greater than 80%) can be obtained, for example, by mutating the sequence during chemical synthesis, followed by the functions described herein. The analysis of the functions retained by the altered macrocyclic peptide (i.e., the functions described in (a) to (i) above) was tested. The biological and/or functional activity of the variant macrocyclic peptide amino acid sequence may be at least about 1×, 2×, 3×, 4×, 5×, 6× of the reference macrocyclic peptide on which the variant is based. 7×, 8×, 9× or 10×. In this context, the term "about" should be interpreted to mean between 0.1 x, 0.2 x, 0.3 x, 0.4 x, 0.5 x, 0.6 x, 0.7 x, 0.8 x or 0.9 x greater or less than the stated amount. Any value.

As used herein, the percent homology between two amino acid sequences corresponds to the percent identity between the two sequences. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (ie, % homology), taking into account the number of gaps and the length of each gap that need to be introduced for optimal alignment of the two sequences. = number of consistent positions / total number of positions × 100). Sequence comparison and determination of the percent identity between two sequences can be accomplished using a mathematical algorithm, as described below in the non-limiting examples.

The percent identity between the two amino acid sequences can be determined using the algorithm of Meyers E. et al. ( Comput. Appl. Biosci. 4: 11-17 (1988)) (this algorithm has been incorporated into the ALIGN program (version 2.0)) Determined using the PAM120 weight residue table, the gap length penalty of 12, and the gap penalty of 4. In addition, the percent identity between the two amino acid sequences can be used by Needleman et al. ( J. Mol. Biol. 48) which has been incorporated into the GAP program of the GCG® kit software (available at www.gcg.com) . 444-453 (1970)) Algorithm, using Blossum 62 matrix or PAM250 matrix, and gap weights 16, 14, 12, 10, 8, 6, or 4 and length weights 1, 2, 3, 4, 5, or 6.

Conservatively modified macrocyclic peptide

In still another embodiment, the macrocyclic peptide of the invention comprises an amino acid sequence homologous to the amino acid sequence of the macrocyclic peptide described herein, and wherein the macrocyclic peptide retains the desired function of the macrocyclic peptide of the invention and/ Or biological characteristics.

For example, the invention provides a macrocyclic peptide or antigen binding portion thereof comprising: an amino acid sequence at least 80% homologous to an amino acid sequence selected from the macrocyclic peptides described herein, wherein one or more amine groups Conservative amino acid has been substituted with an acid; and macrocyclic peptides exhibit one or more of the following characteristics: (a) macrocyclic peptides 200nM 200nM or less than the IC ₅₀ binding to human PD-L1; (b) a macrocyclic peptide does not substantially bind To human CD28, CTLA-4 or ICOS; (c) macrocyclic peptide enhances secretion of IFNγ by CMV-specific T cells; (d) macrocyclic peptide enhances secretion of IFNγ by HIV-specific T cells; (e) macrocyclic peptide binds to human PD -L1 and one or more of the following: cynomolgus PD-L1, woodchuck PD-L1 and/or mouse PD-L1; (f) macrocyclic peptide inhibits PD-L1 and/or PD-L2 binding to PD -1; (g) macrocyclic peptides are capable of competitively binding to anti-PD-1 monoclonal antibodies (including nivozumab (BMS-936558, MDX-1106)); (h) macrocyclic peptides in cell analysis and/or live Inhibition of tumor cell growth in in vivo assays; and/or (i) macrocyclic peptides inhibit HIV in cellular assays and/or in vivo assays.

As used herein, the term "conservative sequence modification" means an amino acid modification that does not significantly affect or alter the binding characteristics of a macrocyclic peptide containing an amino acid sequence. Such conservative modifications include amino acid substitutions, additions and deletions. Modifications can be introduced into the antibodies of the invention by standard techniques known in the art, such as substitution of amidoxime during chemical synthesis, site-directed mutagenesis, and PCR-mediated mutation induction. Conservative amino acid substitutions are those in which the amino acid residue is replaced with an amino acid having a similar side chain amino acid residue. Amino acid residues with similar side chains The family is defined in this technology. Such families include amino acids having the following side chains: basic side chains (eg, amino acid, arginine, histidine), acidic side chains (eg, aspartic acid, glutamic acid), uncharged Polar side chains (eg glycine, aspartic acid, glutamic acid, serine, threonine, tyrosine, cysteine, tryptophan), non-polar side chains (eg propylamine) Acid, valine, leucine, isoleucine, valine, phenylalanine, methionine, beta branched side chains (eg, sulphate, valine, isoleucine) and aromatic Side chains (eg tyrosine, phenylalanine, tryptophan, histidine). Thus, one or more amino acid residues within the antigen binding region of a macrocyclic peptide of the invention can be replaced by other amino acid residues of the same side chain family and using the functional assays described herein for altered antibodies The reserved functions (i.e., the functions described in (a) to (i) above) are tested. Conservative amino acid substitutions may also be selected from one or more of the non-naturally occurring amino acids disclosed herein.

Pharmaceutical composition

The invention further relates to a polypeptide as described herein, wherein the C-terminus and/or N-terminus of the sequence comprises one or more amino acid deletions.

In a preferred embodiment, the present invention comprises a compound N-terminal deletion polypeptide of No. 99: Compound No. 99 X1-X13, X2-X13, X3-X13, X4-X13, X5-X13, X6-X13, X7-X13, X8-X13, X9-X13, X10-X13, X11-X13 and/or X12-X13, wherein each X represents an amino acid at a specified position for each peptide as outlined herein. The invention also encompasses loop forms of such deletion mutants formed using the ligation chemistry described elsewhere herein.

In a preferred embodiment, the invention comprises a C-terminal deletion polypeptide of the compound No. 99: X-X13, X1-X12, X1-X11, X1-X10, X1-X9, X1-X8 of the compound No. 99, X1-X7, X1-X6, X1-X5, X1-X4 and/or X1-X3, wherein each X represents an amino acid at a specified position for each peptide as outlined herein. The invention also encompasses loop forms of such deletion mutants formed using the ligation chemistry described elsewhere herein.

In a preferred embodiment, the N-terminal end of the inclusive compound of the present invention is more than 1 Peptide: Compound No. 1 X1-X15, X2-X15, X3-X15, X4-X15, X5-X15, X6-X15, X7-X15, X8-X15, X9-X15, X10-X15, X11-X15 And/or X12-X15, wherein each X represents an amino acid at a specified position for each peptide as outlined herein. The invention also encompasses loop forms of such deletion mutants formed using the ligation chemistry described elsewhere herein.

In a preferred embodiment, the invention comprises a C-terminal deletion polypeptide of No. 1 of the compound: X1-X15, X1-X14, X1-X13, X1-X12, X1-X11, X1-X10 of the compound No. 1. X1-X9, X1-X8, X1-X7, X1-X6, X1-X5, X1-X4 and/or X1-X3, wherein each X represents an amino acid at a specified position for each peptide as outlined herein. The invention also encompasses loop forms of such deletion mutants formed using the ligation chemistry described elsewhere herein.

In a preferred embodiment, the invention comprises a compound N-terminal deletion polypeptide of No. 71: Compound No. 71, X1-X14, X2-X14, X3-X14, X4-X14, X5-X14, X6-X14, X7-X14, X8-X14, X9-X14, X10-X14, X11-X14 and/or X12-X14, wherein each X represents an amino acid at a specified position for each peptide as outlined herein. The invention also encompasses loop forms of such deletion mutants formed using the ligation chemistry described elsewhere herein.

In a preferred embodiment, the invention comprises a C-terminal deletion polypeptide of the compound No. 71: X-X14, X1-X13, X1-X12, X1-X11, X1-X10, X1-X9 of the compound No. 71, X1-X8, X1-X7, X1-X6, X1-X5, X1-X4 and/or X1-X3, wherein each X represents an amino acid at a specified position for each peptide as outlined herein. The invention also encompasses loop forms of such deletion mutants formed using the ligation chemistry described elsewhere herein.

In another aspect, the invention provides a composition, such as a pharmaceutical composition, comprising one or a combination of a macrocyclic peptide (or antigen binding portion thereof) of the invention formulated together with a pharmaceutically acceptable carrier. Such compositions may include one or a combination of the macrocyclic peptides of the invention (eg two or more different macrocyclic peptides), or immunoconjugates or bispecific molecules. For example, a pharmaceutical composition of the invention may comprise a macrocyclic peptide combination (or immunoconjugate or bispecific) that binds to a different epitope on a target antigen or has a complementary activity.

The pharmaceutical compositions of the invention may also be administered in combination therapy (i.e., in combination with other agents). For example, combination therapies can include a combination of a macrocyclic peptide and at least one other anti-inflammatory or immunosuppressive agent. Examples of therapeutic agents that can be used in combination therapy are described in more detail in the use section of the macrocyclic peptides of the invention.

As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents that are physiologically compatible, and Its analogues. Preferably, the carrier is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., injection or infusion). Depending on the route of administration, the active compound (i.e., macrocyclic peptide, immunoconjugate or bispecific molecule) can coat the material to protect the acid from the action of the acid and other natural conditions that may render the compound inactive.

The pharmaceutical compounds of the invention may include one or more pharmaceutically acceptable salts. "Pharmaceutically acceptable salt" means a salt that retains the desired biological activity of the parent compound and does not produce any undesirable toxicological effects (see, for example, Berge, SM et al, J. Pharm. Sci. , 66:1). -19 (1977)). Examples of such salts include acid addition salts and base addition salts. Acid addition salts include those derived from non-toxic inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, phosphorous acid and the like; non-toxic organic acids such as aliphatic Monocarboxylic acids and dicarboxylic acids, phenyl substituted alkanoic acids, hydroxyalkanoic acids, aromatic acids, aliphatic and aromatic sulfonic acids, and the like. Base addition salts include those derived from alkaline earth metals such as sodium, potassium, magnesium, calcium and the like; and non-toxic organic amines such as N,N'-dibenzylethylenediamine, N - Methylglucosamine, chloroprocaine, choline, diethanolamine, ethylenediamine, procaine and the like.

The pharmaceutical compositions of the present invention may also include a pharmaceutically acceptable antioxidant. medical Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants such as ascorbic acid, cysteamine hydrochloride, sodium hydrogen sulfate, sodium metabisulfite, sodium sulfite, and the like; (2) oil Soluble antioxidants such as ascorbyl palmitate, butylated hydroxymethoxybenzene (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol and the like; (3) Metal chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

Examples of suitable aqueous and non-aqueous vehicles which can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like) and suitable mixtures thereof; vegetable oils, such as Olive oil; and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of a coating material such as lecithin, by the maintenance of the desired particle size in the case of dispersion and by the use of surfactants.

These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the presence of microorganisms can be ensured by the above sterilization procedure and by the inclusion of various antibacterial and antifungal agents (e.g., parabens, chlorobutanol, phenol, sorbic acid, and the like). It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like, in the compositions. In addition, injectable pharmaceutical forms may achieve extended absorption by the inclusion of delayed absorption agents such as aluminum monostearate and gelatin.

The pharmaceutically acceptable carrier includes sterile aqueous solutions or dispersions and sterile powders for the preparation of sterile injectable solutions or dispersions. The use of such media and agents for pharmaceutically active substances is known in the art. Unless any conventional media or agent is incompatible with the active compound, its use in the pharmaceutical compositions of the present invention is contemplated. Supplementary active compounds can also be incorporated into the compositions.

Therapeutic compositions typically must be sterile and stable under the conditions of manufacture and storage. The compositions may be formulated as solutions, microemulsions, liposomes or other ordered structures suitable for high drug concentrations. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. Proper fluidity can be maintained, for example, by the use of a coating material such as lecithin, by the maintenance of the desired particle size in the case of dispersion and by the use of surfactants. In many cases, it is preferred to include an isotonic agent, such as a sugar, a polyol (such as mannitol, sorbitol) or sodium chloride. Injectable compositions can achieve prolonged absorption by including a delayed absorbent such as monostearate and gelatin in the compositions.

Sterile injectable solutions can be prepared by incorporating, as needed, the active compound of the active ingredient in a suitable solvent in one or a combination of the ingredients listed above, followed by microfiltration sterilization. In general, the dispersion is prepared by incorporating the active compound into a sterile vehicle which comprises a basic dispersion medium and other essential ingredients selected from the ingredients listed above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred preparation methods are vacuum drying and lyophilization (lyophilization) to produce the active ingredient plus any other desired ingredient powder from the prior sterile filtration solution.

The amount of active ingredient which may be combined with carrier materials to produce a single dosage form will vary depending upon the subject being treated and the particular mode of administration. The amount of active ingredient which may be combined with carrier materials to produce a single dosage form is usually the amount of the composition which produces a therapeutic effect. Generally, in an amount of from 100%, the amount ranges from about 0.01% to about 99% active ingredient, preferably from about 0.1% to about 70%, optimally from about 1% to about 30%, and A pharmaceutically acceptable carrier combination.

The dosage regimen is adjusted to provide the optimal desired response (e.g., a therapeutic response). For example, a single bolus can be administered, several divided doses can be administered over time, or the dose can be lowered or increased proportionally, depending on the urgency indication of the therapeutic situation. It is especially advantageous to formulate parenteral compositions in unit dosage form for ease of administration and uniform administration. Dosage unit form, as used herein, refers to physically discrete units in a single dosage form suitable for the subject to be treated, each unit containing a predetermined amount of active compound calculated to produce the desired therapeutic effect, and the desired pharmaceutical carrier. The specification of the unit dosage form of the present invention is determined by the following and is directly dependent on (a) the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and (b) compounding the active compound Limitations of individual treatment sensitivities inherent in the technology of matter.

When a macrocyclic peptide is administered, the dosage range is from about 0.0001 mg to 100 mg per kg of host body weight, and more typically from 0.01 mg to 5 mg per kg of host body weight. For example, the dose may be 0.3 mg per kg body weight, 1 mg per kg body weight, 3 mg per kg body weight, 5 mg per kg body weight or 10 mg per kg body weight, or in the range of 1 mg to 10 mg per kg body weight. An exemplary treatment regimen requires administration once a day, twice a week, three times a week, once a week, once every two weeks, once every three weeks, once every four weeks, once a month, once every three months, or every three months Once every 6 months. A preferred dosing regimen of the macrocyclic peptide of the present invention comprises intravenous administration, 1 mg per kilogram of body weight or 3 mg per kilogram of body weight, wherein the antibody is administered using one of the following administration schedules: (i) six times per four weeks , then every three months; (ii) every three weeks; (iii) once every 3 kg of body weight, followed by 1 mg per kilogram of body weight every three weeks.

In some methods, two or more macrocyclic peptides having different binding specificities are administered simultaneously, in which case the administration dose of each compound falls within the specified range. Compounds are usually administered in a variety of settings. The interval between single administrations can be, for example, weekly, monthly, every three months or annually. The interval may also be irregular according to the measurement of the blood content of the macrocyclic peptide for the target antigen in the patient. In some methods, the dosage is adjusted to achieve a plasma antibody concentration of about 1-1000.mu.g/ml and in some methods about 25-300.mu.g/ml.

Alternatively, the macrocyclic peptide can be administered as a sustained release formulation, in which case the frequency of administration is less. The dose and frequency vary depending on the half-life of the macrocyclic peptide in the patient. The dose and frequency of administration vary depending on whether the treatment is preventive or therapeutic. In prophylactic applications, relatively low doses are administered at relatively infrequent intervals over a longer period of time. Some patients continue to receive treatment for the rest of their lives. In therapeutic applications, it is sometimes desirable to administer relatively high doses at relatively short intervals until disease progression is reduced or terminated, preferably until the patient shows partial or complete improvement in disease symptoms. Thereafter, a prophylactic regimen can be administered to the patient.

The actual dosage level of the active ingredient in the pharmaceutical compositions of the present invention can be varied to achieve an amount of active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without poisoning the patient. The selected dosage level will depend on a variety of pharmacokinetic factors, including the activity of the particular composition of the invention or its ester, salt or guanamine; the route of administration; the time of administration; the rate of excretion of the particular compound employed; the duration of treatment; Other drugs, compounds and/or substances used in combination with the composition; age, sex, weight, condition, general health and prior medical history of the patient to be treated; and similar factors well known in the medical arts.

The "therapeutically effective dose" of the macrocyclic peptide of the present invention preferably lowers the severity of the symptoms of the disease, increases the frequency and duration of the period of the disease-free condition, or prevents dysfunction or disability due to the disease. For example, in the treatment of a tumor, the "therapeutically effective dose" preferably inhibits cell growth or tumor growth by at least about 20%, more preferably at least about 40%, and even more preferably at least about 60, relative to the untreated individual. %, and still better inhibit at least about 80%. The ability of a compound to inhibit tumor growth and/or HIV can be assessed using an animal model system that predicts efficacy or viral efficacy against human tumors. Alternatively, this property of the composition can be assessed by examining the compound's ability to inhibit, which can be examined by in vitro analysis known to the skilled practitioner. A therapeutically effective amount of a therapeutic compound can reduce tumor size, reduce viral load, or otherwise ameliorate symptoms in an individual. Those of ordinary skill in the art will be able to determine such amounts based on factors such as the size of the individual, the severity of the individual's symptoms, and the particular composition or route of administration selected.

In another aspect, the invention provides a dispensing portion of a medical kit comprising a macrocyclic peptide and an anti-CTLA-4 antibody, as described herein. The kit may further comprise instructions for treating a proliferative disease, such as a cancer described herein, and/or an antiviral disease.

The compositions of the present invention can be administered via one or more routes of administration, using one or more of a variety of methods known in the art. As will be appreciated by those skilled in the art, the route and/or mode of administration will vary depending on the desired result. Preferred route for the macrocyclic peptide of the present invention Pathways include intravenous, intramuscular, intradermal, intraperitoneal, subcutaneous, spinal or other parenteral routes of administration such as injection or infusion. As used herein, the phrase "parenteral administration" means a mode of administration other than enteral and topical administration, generally by injection, and includes (not limited to) intravenous, intramuscular, intraarterial, intrathecal, sac Internal, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions.

Alternatively, the macrocyclic peptides of the invention may be administered via a parenteral route, such as a topical, epidermal or mucosal route of administration, such as intranasal, oral, vaginal, rectal, sublingual or topical.

The active compounds can be prepared with carriers that protect the compound against rapid release, such as controlled release formulations, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Many methods for preparing such formulations are patented or generally known to those skilled in the art. See, for example, Robinson, JR, eds., Sustained and Controlled Release Drug Delivery Systems , Marcel Dekker, Inc., New York (1978).

Therapeutic compositions can be administered using medical devices known in the art. For example, in a preferred embodiment, the therapeutic compositions of the present invention can be administered using a needleless hypodermic injection device, such as U.S. Patent Nos. 5,399,163, 5,383,851, 5,312,335, 5,064,413, 4,941,880. The device disclosed in 4,790,824 or 4,596,556. Examples of well-known implants and modules that are suitable for use in the present invention include: U.S. Patent No. 4,487,603, the disclosure of which is incorporated herein by reference in its entirety in its entirety the entire entire entire entire entire entire entire entire disclosure A therapeutic device for a drug; US Patent No. 4, 447, 233, which discloses a drug infusion pump that delivers a drug at an accurate infusion rate; U.S. Patent No. 4,447,224, which is incorporated herein by reference. It discloses a osmotic drug delivery system having a multi-chamber metabolic zone; and U.S. Patent No. 4,475,196, which discloses a permeable drug delivery system. These patents are incorporated herein by reference. Many other such implants, delivery systems and modules are known to those skilled in the art.

In certain embodiments, the macrocyclic peptides of the invention can be formulated to ensure proper in vivo distribution. For example, the blood brain barrier (BBB) rejects many highly hydrophilic compounds. To ensure that the therapeutic compounds of the invention pass through the BBB (if desired), they can be formulated, for example, as liposomes. For a method of producing a liposome, see, for example, U.S. Patent Nos. 4,522,811, 5,374,548, and 5,399,331. Liposomes can comprise one or more moieties that are selectively delivered into a particular cell or organ to enhance targeted drug delivery (see, eg, Ranade, VV, J. Clin. Pharmacol. , 29:685 (1989)). Illustrative targeting moieties include folate or biotin (see, e.g., U.S. Patent No. 5,416,016 issued to Low et al.); Mannose (Umezawa et al. , Biochem. Biophys. Res. Commun. , 153: 1038 (1988) Macrocyclic peptides (Bloeman, PG et al, FEBS Lett. , 357: 140 (1995); Owais, M. et al., Antimicrob. Agents Chemother. , 39: 180 (1995)); surfactant A (Briscoe et al, Am. J. Physiol. , 1233: 134 (1995)); p120 (Schreier et al, J. Biol. Chem. , 269: 9090 (1994)); see also Keinanen, K. et al. , FEBS Lett. , 346: 123 (1994); Killion, JJ et al, Immunomethods 4: 273 (1994).

Use and method of the present invention

The macrocyclic peptides, compositions and methods of the invention have a number of in vitro and in vivo utilities, including, for example, detecting PD-L1 or enhancing the immune response by blocking PD-L1. For example, such molecules can be administered to cells in culture in vitro or ex vivo, or to human subjects, such as in vivo, to human subjects to enhance immunity in a variety of situations. Thus, in one aspect, the invention provides a method of modulating an immune response in an individual comprising administering to the individual an antibody or antigen binding portion thereof of the invention to modulate the immune response of the individual. It is preferred to enhance, stimulate or upregulate the response. In other aspects, the macrocyclic peptide can have anti-cynomolgus, anti-mouse and/or anti-mouse control and therapeutic activity.

As used herein, the term "individual" is intended to include both human and non-human animals. Non-human animals include all vertebrates, such as mammals and non-mammals, such as non-human primates, sheep, dogs, cats, cows, horses, chickens, groundhogs, amphibians and reptiles, but mammals. Preferably, such as non-human primates, sheep, dogs, cats, cows and horses. Preferred individuals include human patients who need to enhance the immune response. These methods are particularly suitable for treating human patients suffering from conditions which can be treated by enhancing T cell mediated immune responses. In particular embodiments, the methods are particularly suitable for treating cancer cells in vivo. To achieve antigen-specific enhancement of immunity, macrocyclic peptides can be administered with the antigen of interest. When a macrocyclic peptide directed against PD-L1 is administered with another agent, the two may be administered sequentially or simultaneously.

The invention further provides for detecting the presence of human, marmot, cynomolgus and/or mouse PD-L1 antigen in a sample or measuring human, woodchuck, cynomolgus and/or mouse PD-L1 antigen A method comprising the step of binding a sample and a control sample to a reference monoclonal antibody or antigen-binding portion thereof that specifically binds to humans, woodchucks, cynomolgus monkeys and/or PD-L1, in allowing the antibody or a portion thereof to be human, Contact between the groundhog, cynomolgus monkey and/or mouse PD-L1 under conditions of formation of a complex. The formation of the complex is then detected, wherein a difference in complex formation between the samples compared to the control sample indicates the presence of human, woodchuck, cynomolgus and/or mouse PD-L1 antigen in the sample.

Compared with CD28, ICOS and CTLA-4, if the macrocyclic peptide of the present invention specifically binds to PD-L1, the macrocyclic peptide of the present invention can be used for specifically detecting PD-L1 expression on the cell surface, and in addition, Purification by immunoaffinity for purification of PD-L1.

cancer

Macrocyclic peptides block PD-1 to enhance the patient's immune response against cancer cells. The ligand PD-L1 of PD-1 is not expressed in normal human cells, but is abundant in various human cancers (Dong et al. , Nat. Med. , 8: 787-789 (2002)). The interaction between PD-1 and PD-L1 reduces tumor infiltrating lymphocytes, reduces T cell receptor-mediated proliferation, and immune evasion of cancer cells (Dong et al, J. Mol. Med. , 81:281- 287 (2003); Blank et al, Cancer Immunol. Immunother . , 54: 307-314 (2005); Konishi et al, Clin. Cancer Res. , 10: 5094-5100 (2004)). Immune suppression can be reversed by inhibiting the local interaction between PD-1 and PD-L1, and this reversal is additive when the interaction between PD-1 and PD-L2 is also blocked (Iwai et al. , Proc. Natl. Acad. Sci. , 99: 12293-12297 (2002); Brown et al, J. Immunol. , 170: 1257-1266 (2003)). Although previous studies have shown that T cell proliferation can be repaired by inhibiting the interaction of PD-1 with PD-L1, the direct effect of blocking PD-1/PD-L1 interaction on cancer tumor growth has not been reported. In one aspect, the invention relates to the in vivo treatment of a subject using a macrocyclic peptide that inhibits the growth of a cancerous tumor. Macrocyclic peptides can be used alone to inhibit cancer tumor growth. Alternatively, macrocyclic peptides can be used in combination with other immunogenic agents, standard cancer therapies, or other macrocyclic peptides, as described below.

Accordingly, in one embodiment, the invention provides a method of inhibiting tumor cell growth in an individual comprising administering to the individual a therapeutically effective amount of a macrocyclic peptide or antigen binding portion thereof.

Preferred cancers that can inhibit growth using the macrocyclic peptides of the invention include those that are typically responsive to immunotherapy. Non-limiting examples of treatable cancers include melanoma (metastatic melanoma), renal cell carcinoma (eg, clear cell carcinoma), prostate cancer (eg, hormone-refractory prostate cancer and castration-refractory prostate cancer), breast cancer, Colorectal cancer and lung cancer (eg, squamous and non-squamous non-small cell lung cancer). Additionally, the invention encompasses refractory or relapsing malignant diseases that inhibit growth using the macrocyclic peptides of the invention.

Other cancers that may be treated using the methods of the invention include bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular malignant melanoma, uterine cancer, ovarian cancer, colon cancer, rectal cancer, anal cancer, Gastric cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's Disease, non-Hodgkin's lymphoma, esophageal cancer, small intestine cancer, Endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, chronic or acute leukemia (including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic Leukemia), childhood solid tumors, lymphocytic lymphoma, bladder cancer, renal or ureteral cancer, renal pelvic cancer, central nervous system (CNS) neoplasm, primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, Brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell carcinoma, T-cell lymphoma, environmental inducement Cancer, including asbestos-induced cancer, and combinations of such cancers. The invention is also applicable to the treatment of metastatic cancer, especially metastatic cancers that exhibit PD-L1 (Iwai et al, Int. Immunol. , 17: 133-144 (2005)).

Macrocyclic peptides directed against PD-L1 may optionally be combined with immunogenic agents, such as cancer cells, purified tumor antigens (including recombinant proteins, peptides and carbohydrate molecules), cells, and gene transgenes encoding immunostimulatory cytokines Dyed cells (He et al. , J. Immunol. , 173:4919-4928 (2004)). Non-limiting examples of tumor vaccines that can be used include peptides of melanoma antigens, such as gp100, MAGE antigen, Trp-2, MART1 and/or tyrosinase peptides, or tumors transfected to express the cytokine GM-CSF Cells (discussed further below).

Some tumors in humans have been shown to be immunogenic, such as melanoma. It is foreseeable that by increasing the T cell activation threshold by a PD-L1 blocker, it is expected that the tumor response in the host will be activated.

PD-L1 blockers may be most effective when combined with a vaccination regimen. Many experimental vaccination strategies for tumors have been designed (see Rosenberg, S., Development of Cancer Vaccines, ASCO Educational Book Spring: 60-62 (2000); Logothetis, C., ASCO Educational Book Spring: 300-302 ( 2000); Khayat, D., ASCO Educational Book Spring: 414-428 (2000); Foon, K., ASCO Educational Book Spring: 730-738 (2000); see also Restifo, N. et al., Cancer Vaccines, Chapter 61, pp. 3023-3043, edited by DeVita, V. et al., Cancer: Principles and Practice of Oncology , 5th Ed. (1997)). In one of these strategies, vaccines are prepared using autologous or allogeneic tumor cells. These cell vaccines have been shown to be most effective when tumor cells are transduced to express GM-CSF. In tumor vaccination, GM-CSF has been shown to be a strong activator of antigen presentation (Dranoff et al, Proc. Natl. Acad. Sci. USA , 90: 3539-3543 (1993)).

Studies of gene expression and large-scale gene expression patterns in various tumors have produced definitions of so-called tumor-specific antigens (Rosenberg, SA, Immunity , 10:281-287 (1999)). In many cases, such tumor-specific antigens are differentiation antigens expressed in cells from which tumors and tumors are derived, such as melanocyte antigen gp100, MAGE antigen, and Trp-2. More importantly, many of these antigens are shown to be targets of tumor-specific T cells found in the host. The PD-L1 blocker can be used in conjunction with a number of recombinant proteins and/or peptides expressed in tumors to produce an immune response against such proteins. These proteins are often recognized by the immune system as autoantigens and are therefore tolerant to them. Tumor antigens may also include protein telomerase, which is required for the synthesis of telomeres and is expressed in more than 85% of human cancers and in only limited somatic tissues (Kim, N. et al., Science , 266). :2011-2013 (1994)). (The somatic tissues can be protected in various ways to prevent immune attack). Tumor antigens may also be due to changes in protein sequences or somatic mutations that result in fusion proteins between the two unrelated sequences (ie, bcr-abl in the Philadelphia chromosome), or individual genotypes from B cell tumors. It is a "new antigen" expressed in cancer cells.

Other tumor vaccines may include proteins involved in human cancer viruses, such as human papillomavirus (HPV), hepatitis virus (HBV and HCV), and Kaposi's herpes sarcoma virus (KHSV). Another form of tumor-specific antigen that can be used in conjunction with a PD-L1 blocker is purified heat shock protein (HSP) isolated from tumor tissue itself. Such heat shock proteins contain fragments of proteins from tumor cells and such HSPs are efficiently delivered to antigen presenting cells for eliciting tumor immunity (Suot, R. et al, Science , 269: 1585-1588 (1995); Tamura, Y Et al., Science , 278: 117-120 (1997)).

Dendritic cells (DCs) are strong antigen presenting cells that can be used to sensitize antigen-specific responses. DCs can be produced ex vivo and loaded with various protein and peptide antigens as well as tumor cell extracts (Nestle, F. et al, Nat. Med. , 4: 328-332 (1998)). DCs can also be transduced by genetic means to also express such tumor antigens. DC has also been directly fused to tumor cells for immunization purposes (Kugler, A. et al, Nat. Med. , 6: 332-336 (2000)). In vaccination methods, DC immunization can be effectively combined with a PD-L1 blocker to activate a stronger anti-tumor response.

PD-L1 blockers can also be combined with standard cancer therapies. The PD-L1 blocker can be effectively combined with a chemotherapeutic regimen. In such cases, it is possible to reduce the dosage of the chemotherapeutic agent (Mokyr, M. et al, Cancer Res. , 58: 5301-5304 (1998)). An example of this combination is the combination of a macrocyclic peptide and dacarbazine for the treatment of melanoma. Another example of this combination is the combination of a macrocyclic peptide and interleukin-2 (IL-2) for the treatment of melanoma. The scientific rationale behind the use of PD-L1 blockers in combination with chemotherapy is that cell death, which is a consequence of the cytotoxic effects of most chemotherapeutic compounds, should result in increased levels of tumor antigens in the antigen presentation pathway. Other combination therapies that can synergize with PD-L1 blockers via cell death are radiation, surgery, and hormone removal. Each of these protocols produces a source of tumor antigen in the host. Angiogenesis inhibitors can also be combined with PD-L1 blockers. Inhibition of angiogenesis causes tumor cell death so that tumor antigens can be fed into the host antigen presentation pathway.

The PD-L1 blocking macrocyclic peptide can also be used in combination with a bispecific macrocyclic peptide that targets an effector cell expressing an Fcα or Fcγ receptor to a tumor cell (see, for example, U.S. Patent Nos. 5,922,845 and 5,837,243). Bispecific macrocyclic peptides can be used to target two separate antigens. For example, anti-Fc receptor/anti-tumor antigen (eg, Her-2/neu) bispecific macrocyclic peptides have been used to target macrophages to tumor sites. This targeting can activate tumor-specific responses more efficiently. The use of PD-L1 blockers enhances the T cell arms of these responses. Alternatively, by using a bispecific macrocyclic peptide that binds to tumor antigens and dendritic cell-specific cell surface markers The antigen is delivered directly to the DC.

Tumors evade host immune surveillance through a variety of mechanisms. Inactivation of proteins that are manifested by tumors and immunosuppressive can overcome many of these mechanisms. These mechanisms include, inter alia, TGF-β (Kehrl, J. et al. , J. Exp. Med. , 163: 1037-1050 (1986)), IL-10 (Howard, M. et al., Immunology Toddy , 13:198). -200 (1992)) and Fas ligand (Hahne, M. et al., Science , 274: 1363-1365 (1996)). Macrocyclic peptides for each of these entities can be used in combination with anti-PD-L1 to counteract the effects of immunosuppressive agents and contribute to the host's tumor immune response.

Other macrocyclic peptides that can be used to activate host immunoreactivity can be used in combination with anti-PD-L1. Such other macrocyclic peptides include molecules that activate DC function and antigen presentation on the surface of dendritic cells. Anti-CD40 macrocyclic peptides are effective in replacing T cell-assisted activity (Ridge, J. et al., Nature , 393: 474-478 (1998)) and can be used in conjunction with PD-1 antibodies (Ito, N. et al., Immunobiology , 201(5): 527-540 (2000)). Activation of macrocyclic peptides against T cell costimulatory molecules such as the following may also increase the degree of T cell activation: CTLA-4 (e.g., U.S. Patent No. 5,811,097), OX-40 (Weinberg, A. et al., Immunol. , 164). : 2160-2169 (2000)), 4-1BB (Melero, I. et al. , Nat. Med. , 3: 682-685 (1997) and ICOS (Hutloff, A. et al., Nature , 397: 262-266) (1999)).

Bone marrow transplantation is currently used to treat a variety of hematopoietic tumors. Although graft versus host disease is the result of this treatment, therapeutic benefit can be obtained from the graft anti-tumor response. PD-L1 blockers can be used to increase the effectiveness of donor-implanted tumor-specific T cells.

There are also several experimental treatment options, including ex vivo activation and expansion of antigen-specific T cells and their subsequent inheritance transfer to recipients in order to target antigen-specific T cells to tumors (Greenberg, R. et al. , Science , 285: 546-551 (1999)). These methods can also be used to activate T cell responses to infectious agents such as CMV. Ex vivo activation in the presence of macrocyclic peptides is expected to increase the frequency and activity of inherited metastatic T cells.

Infectious disease

Other methods of the invention are used to treat patients who have been exposed to a particular toxin or pathogen. Accordingly, another aspect of the invention provides a method of treating an infectious disease in an individual comprising administering to the individual a macrocyclic peptide of the invention or an antigen binding portion thereof, in order to treat an infectious disease in the subject. Preferably, the antibody is a human anti-human PD-L1 macrocyclic peptide (such as any of the macrocyclic peptides described herein). Additionally or alternatively, the antibody can be a chimeric or humanized antibody.

As discussed above, similar to the use of tumors, antibody-mediated PD-L1 blockers can be used alone or as an adjuvant in combination with a vaccine to stimulate an immune response against pathogens, toxins, and autoantigens. Examples of pathogens to which this treatment is particularly applicable include pathogens that currently have no effective vaccine, or pathogens that are not fully effective vaccines. Such pathogens include, but are not limited to, HIV, hepatitis (A, B, and C), influenza, herpes, Giardia, malaria (Butler, NS et al, Nature Immunology, 13 : 188-195 ( 2012); Hafalla, JCR et al., PLOS Pathogens (February 2, 2012)), Leishmania, Staphylococcus aureus, Pseudomonas Aeruginosa. PD-L1 blockers are particularly useful for infections established by vehicles such as HIV, which present altered antigens during infection. These novel epitopes are recognized as foreign when administered against human PD-L1, thereby eliciting a strong T cell response that is not attenuated by the negative signal of PD-L1.

Some examples of infectious pathogenic viruses that can be treated by the methods of the invention include HIV, hepatitis (A, B or C), herpes viruses (eg, VZV, HSV-1, HAV-6, HSV-II, and CMV, Ai-Ba II) Epstein Barr virus, adenovirus, influenza virus, flavivirus, echovirus, rhinovirus, coxsackie virus, coronavirus, respiratory syncytial virus, mumps virus, Rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, soft prion, poliovirus, rabies virus, JC virus and arbovirus viral encephalitis virus.

Some examples of infectious pathogenic bacteria that can be treated by the methods of the invention include Chlamydia, rickettsial bacteria, Mycobacterium, Staphylococcus, Streptococcus, Pneumonococci, Meningitis Meningococci and conococci, klebsiella, proteus, serratia, pseudomonas, legionella, diphtheria, salmonella, bacilli, cholera, Tetanus, botulism, anthrax, plague, leptospirosis and Lyme disease.

Infectious pathogenic fungi which can be treated by the method of the present invention include Candida (albicans, krusei, glabrata, tropicalis) Etc., Cryptococcus neoformans, Aspergillus (Fumigatus, Niger, etc.), Mucorales (Muscularus, Mucor, Abalone) Absidia), rhizophus, Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis and pods Histoplasma capsulatum.

Some examples of infectious pathogenic parasites that can be treated by the methods of the invention include Entamoeba histolytica, Balantidium coli, Naegleria fowleri, Acanthamoeba Actanthamoeba sp., Giardia lambia, Cryptosporidium sp., Pneumocystis carinii, Plasmodium vivax, microbar Babesia microti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani, Toxoplasma gondi, and Nippostrongylus Brasiliensis).

In all of the above methods, the PD-L1 blocker can be combined with other forms of immunotherapy, such as cytokine therapy (eg, interferon, targeting VEGF activity or the VEGF receptor GM-CSF, G-CSF, IL-2). a pharmaceutical agent, or a bispecific antibody therapy that enhances the presentation of tumor antigens (see, eg, Holliger, Proc. Natl. Acad. Sci. USA , 90:6444-6448 (1993); Poljak, Structure , 2: 1121-1123 ( 1994)).

Autoimmune response

Macrocyclic peptides can stimulate and amplify autoimmune responses. In fact, the use of tumor cells and peptide vaccines to induce anti-tumor responses revealed that many anti-tumor responses include anti-autoreactivity (de-pigmentation was observed in B16 melanoma with improved anti-CTLA-4+GM-CSF (van Elsas et al., Ibid)) Depigmentation in mice vaccinated with Trp-2 (Overwijk, W. et al, Proc. Natl. Acad. Sci. USA, 96: 2982-2987 (1999)); TRAMP tumor cell vaccine caused by Body immunoprostatitis (Hurwitz, A., supra (2000)); melanoma peptide antigen vaccination and leukoplakia observed in human clinical trials (Rosenberg, SA et al, J. Immunotherm. Emphasis Tumor Immunol. , 19 (1): 81-84 (1996)).

Therefore, the use of anti-PD-L1 blockers in combination with various self proteins can be considered in order to design a vaccination regimen that efficiently produces an immune response against such self proteins in order to treat the disease. For example, Alzheimer's disease involves the improper accumulation of A.beta peptides in amyloid deposits in the brain; antibody responses to amyloid can clear these amyloid deposits ( Schenk et al., Nature , 400: 173-177 (1999)).

Other self proteins can also be used as targets for the treatment of allergies and asthma, such as IgE, and targets for the treatment of rheumatoid arthritis, such as TNFα. Finally, antibody responses to various hormones can be induced using the macrocycles disclosed herein. Neutralizing antibody responses to reproductive hormones can be used for contraception. Neutralizing antibody responses to hormones and other soluble factors can also be considered as possible vaccination targets that are necessary for specific tumor growth.

Similar methods as described above using anti-PD-L1 macrocycles can be used to induce therapeutic autoimmune responses to treat other autoantigens (such as amyloid deposits, including Azhai A patient with impaired accumulation of Aβ, cytokines (such as TNFα) and IgE in Melody's disease.

vaccine

Macrocyclic peptides can be used to stimulate antigen-specific immune responses by co-administering an anti-PD-1 macrocycle with an antigen of interest (eg, a vaccine). Thus, in another aspect, the invention provides a method of enhancing an individual's immune response against an antigen comprising administering to the individual: (i) an antigen; and (ii) an anti-PD-1 macrocycle, or antigen binding thereof Partly to enhance the immune response against the antigen in the individual. The antigen can be, for example, a tumor antigen, a viral antigen, a bacterial antigen, or a pathogen antigen. Non-limiting examples of such antigens include those discussed in the above sections, such as the tumor antigens (or tumor vaccines) discussed above, or antigens from the above viruses, bacteria or other pathogens.

Suitable routes for administering the compositions of the invention (e.g., macrocyclic peptides, multispecific and bispecific molecules, and immunoconjugates) in vivo and ex vivo are well known in the art and can be selected by those of ordinary skill in the art. . For example, the composition can be administered by injection (eg, intravenously or subcutaneously). Suitable dosages of the molecules employed will depend on the age and weight of the individual and the concentration and/or formulation of the composition.

As described previously, the macrocyclic peptides of the invention can be co-administered with one or more other therapeutic agents, such as cytotoxic agents, radiotoxic agents or immunosuppressive agents. The peptide can be attached to the agent (in the form of an immune complex) or can be administered separately from the agent. In the latter case (administered separately), the peptide may be administered before, after or in parallel with the agent, or may be co-administered with other known therapies (e.g., anti-cancer therapies, such as radiation). Such therapeutic agents include, inter alia, anti-neoplastic agents such as doxorubicin (adriamycin), cisplatin/bleomycin sulfate, carmustine ), chlorambucil, decarbazine, and cyclophosphamide/hydroxyurea are themselves effective only at levels that are toxic or subtoxic to the patient. Cisplatin was administered 100 mg per dose intravenously once every four weeks, and doxorubicin was administered intravenously at a dose of 60-75 mg/mL once every 21 days. The macrocyclic peptide of the present invention or antigen-binding fragment thereof is co-administered with a chemotherapeutic agent to make two anticancer drugs The agent acts via different mechanisms to produce a cytotoxic effect on human tumor cells. This co-administration can solve the problem because the development of drug resistance or the change in the antigenicity of the tumor cells makes it non-responsive to the peptide.

Kits comprising compositions of the invention (e.g., macrocyclic peptides, bispecific or multispecific molecules, or immunoconjugates) and instructions for use are also within the scope of the invention. The kit may further comprise at least one other agent, or one or more other macrocyclic peptides of the invention (eg, a human antibody having complementary activity that binds to an epitope in the PD-L1 antigen that is different from the macrocycle). The set typically includes indicia indicating the intended use of the contents of the set. The terminology mark includes any written or recorded material that is supplied on a kit or in a joint kit, or otherwise attached to the kit.

Combination therapy

The macrocyclic peptide of the present invention is useful in combination with another PD-L1 antagonist and/or CTLA-4 antagonist to enhance the immune response against proliferative diseases by blocking PD-L1 and CTLA-4. For example, such molecules can be administered to cells in culture in vitro or ex vivo, or to human subjects, such as in vivo, to human subjects to enhance immunity in a variety of situations. Thus, in one aspect, the invention provides a method of modulating an immune response in an individual comprising administering to the individual a combination of an antibody of the invention or an antigen binding portion thereof to modulate an immune response in the individual. It is preferred to enhance, stimulate or upregulate the response. In another embodiment, the invention provides a method of using an immunostimulatory therapeutic to alter an adverse event associated with treating a proliferative disorder, comprising administering to the subject a macrocyclic peptide of the invention and a subtherapeutic dose of an anti-CTLA-4 antibody .

Macrocyclic peptides block PD-L1 and CTLA-4 to enhance the patient's immune response against cancer cells. Cancers that can inhibit growth using the macrocyclic peptides of the invention include cancers that are typically responsive to immunotherapy. Representative examples of cancers that can be treated by the combination therapies of the invention include melanoma (e.g., metastatic malignant melanoma), kidney cancer, prostate cancer, breast cancer, colon cancer, and lung cancer. Other cancer examples that can be treated using the methods of the invention include bone cancer, pancreatic cancer, and skin Skin cancer, head or neck cancer, skin or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, stomach cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vagina Cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin's lymphoma, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, Chronic or acute leukemia (including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia), childhood solid tumor, lymphocytic lymphoma, bladder cancer, renal or ureteral cancer, renal pelvic cancer , central nervous system (CNS) neoplasm, primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell carcinoma, T cell lymphoma, environmentally induced cancer, including asbestos-induced cancer, and combinations of such cancers. The invention is also applicable to the treatment of metastatic cancer.

In certain embodiments, a therapeutic combination comprising at least one macrocyclic peptide described herein can be administered in a single composition in a pharmaceutically acceptable carrier, or in which each agent can be administered sequentially The individual compositions are administered in parallel. For example, an anti-CTLA-4 antibody and a macrocyclic peptide of the invention can be administered sequentially, such as first administration of anti-CTLA-4 and second administration of a macrocyclic peptide, or first administration of a macrocyclic peptide and second administration of anti-CTLA -4. Furthermore, if more than one dose of combination therapy is administered sequentially, the order of sequential administration may be reversed or maintained in the same order at each administration time point, sequential administration may be combined with concurrent administration, or any combination thereof. For example, the first administration of an anti-CTLA-4 antibody in combination with a macrocyclic peptide can be in parallel, and the second administration can be sequential, wherein anti-CTLA-4 is administered first and then macrocyclic peptide is administered, and The third administration can be sequential, wherein the macrocyclic peptide is administered first and the anti-CTLA-4 or the like is administered second. Another representative dosing regimen can include first administration of the macrocyclic peptide and second administration of the first sequential administration of anti-CTLA-4, and subsequent administration can be in parallel.

Combinations of macrocyclic peptides and anti-CTLA-4 agents can be further combined with immunogenic agents, such as cancer cells, purified tumor antigens (including recombinant proteins, peptides and carbohydrate molecules), cells, and encoded immunostimulatory cells, as appropriate. Hormone gene transfected cells (He et al. , J. Immunol. , 173:4919-4928 (2004)). Non-limiting examples of tumor vaccines that can be used include peptides of melanoma antigens, such as gp100, MAGE antigen, Trp-2, MART1 and/or tyrosinase peptides, or tumors transfected to express the cytokine GM-CSF Cells (discussed further below).

The combined PD-L1 macrocyclic peptide and CTLA-4 blocker can be further combined with a vaccination regimen. Many experimental vaccination strategies for tumors have been designed (see Rosenberg, S., Development of Cancer Vaccines, ASCO Educational Book Spring: 60-62 (2000); Logothetis, C., ASCO Educational Book Spring: 300-302 ( 2000); Khayat, D., ASCO Educational Book Spring: 414-428 (2000); Foon, K., ASCO Educational Book Spring: 730-738 (2000); see also Restifo et al, Cancer Vaccines, Chapter 61, Pp. 3023-3043, edited by DeVita et al., Cancer: Principles and Practice of Oncology , 5th Ed. (1997)). In one of these strategies, vaccines are prepared using autologous or allogeneic tumor cells. These cell vaccines have been shown to be most effective when tumor cells are transduced to express GM-CSF. In tumor vaccination, GM-CSF has been shown to be a strong activator of antigen presentation (Dranoff et al, Proc. Natl. Acad. Sci. USA , 90: 3539-3543 (1993)).

Study of gene expression in a variety of tumors and large-scale gene expression patterns have resulted in the definition of so-called tumor-specific antigens (Rosenberg, Immunity, 10: 281-287 (1999)). In many cases, such tumor-specific antigens are differentiation antigens expressed in cells from which tumors and tumors are derived, such as melanocyte antigen gp100, MAGE antigen, and Trp-2. More importantly, many of these antigens are shown to be targets of tumor-specific T cells found in the host. In certain embodiments, the combined PD-L1 macrocyclic peptide and CTLA-4 blocker using the antibody compositions described herein can be used in conjunction with a number of recombinant proteins and/or peptides expressed in tumors to produce An immune response against these proteins. These proteins are often recognized by the immune system as autoantigens and are therefore tolerant to them. Tumor antigens may also include protein telomerase, which is required for the synthesis of telomeres and is expressed in more than 85% of human cancers and in only limited somatic tissues (Kim et al., Science , 266:2011- 2013 (1994)). (The somatic tissues can be protected in various ways to prevent immune attack). Tumor antigens may also be due to changes in protein sequences or somatic mutations that result in fusion proteins between the two unrelated sequences (ie, bcr-abl in the Philadelphia chromosome), or individual genotypes from B cell tumors. It is a "new antigen" expressed in cancer cells.

Other tumor vaccines may include proteins involved in human cancer viruses, such as human papillomavirus (HPV), hepatitis virus (HBV and HCV), and Kaposi's herpes sarcoma virus (KHSV). Another form of tumor-specific antigen that can be used in conjunction with a PD-L1 macrocyclic peptide blocker is purified heat shock protein (HSP) isolated from tumor tissue itself. Such heat shock proteins contain fragments of proteins from tumor cells and such HSPs are efficiently delivered to antigen presenting cells for eliciting tumor immunity (Suot et al, Science , 269: 1585-1588 (1995); Tamura et al, Science , 278:117-120 (1997)).

Dendritic cells (DCs) are strong antigen presenting cells that can be used to sensitize antigen-specific responses. DCs can be produced ex vivo and loaded with various protein and peptide antigens as well as tumor cell extracts (Nestle et al, Nat. Med. , 4:328-332 (1998)). DCs can also be transduced by genetic means to also express such tumor antigens. DC has also been directly fused to tumor cells for immunization purposes (Kug1er et al, Nat. Med. , 6: 332-336 (2000)). In the vaccination method, DC immunization can be further effectively combined with the combined anti-PD-L1 macrocyclic peptide and CTLA-4 blocker to activate a stronger anti-tumor response.

The combined anti-PD-L1 macrocyclic peptide and CTLA-4 blocker can also be further combined with standard cancer therapies. For example, the combined macrocyclic peptides and CTLA-4 blockers can be effectively combined with a chemotherapeutic regimen. In such cases, as observed in the combination of the macrocyclic peptide and the anti-CTLA-4 agent, the dosage of other chemotherapeutic agents administered in combination with the present invention can be reduced (Mokyr et al, Cancer Res. , 58). :5301-5304 (1998)). An example of such a combination is the combination of a macrocyclic peptide with an anti-CTLA-4 agent and further with a combination of dacarbazine for the treatment of melanoma. Another example is the combination of a macrocyclic peptide with an anti-CTLA-4 agent in combination with interleukin-2 (IL-2) for the treatment of melanoma. The scientific rationale behind the use of PD-L1 macrocyclic peptides and CTLA-4 blockers in combination with chemotherapy is that cell death as a consequence of the cytotoxic effects of most chemotherapeutic compounds should result in the content of tumor antigens in the antigen presentation pathway. improve. Other combination therapies that may be synergistic with cell-deathing by the combined anti-PD-L1 macrocyclic peptide and CTLA-4 blocker include radiation, surgery or hormone removal. Each of these protocols produces a source of tumor antigen in the host. Angiogenesis inhibitors can also be combined with the combined PD-L1 and CTLA-4 blockers. Inhibition of angiogenesis causes tumor cell death, and tumor cell death can also be a source of tumor antigen that is fed into the host antigen presentation pathway.

Combinations of PD-L1 and CTLA-4 blockers can also be used in combination with bispecific macrocyclic peptides that target effector cells expressing Fc[alpha] or Fc[gamma] receptors to tumor cells (see, eg, USA) Patent Nos. 5,922,845 and 5,837,243). Bispecific macrocyclic peptides can be used to target two separate antigens. For example, anti-Fc receptor/anti-tumor antigen (eg, Her-2/neu) bispecific macrocyclic peptides have been used to target macrophages to tumor sites. This targeting can activate tumor-specific responses more efficiently. The combination of PD-1 and CTLA-4 blockers enhances the T cell arm of these responses. Alternatively, the antigen can be delivered directly to DC by using a bispecific macrocyclic peptide that binds to tumor antigen and dendritic cell-specific cell surface markers.

In another example, a combination of a macrocyclic peptide and an anti-CTLA-4 agent can be used in combination with an anti-neoplastic macrocyclic agent, such as RITUXAN® (rituximab), HERCEPTIN® (trastuzumab ( Trastuzumab)), BEXXAR® (tositumomab), ZEVALIN® (ibritumomab), CAMPATH® (alemtuzumab), lymphoblastic agent (Ipa) Bep monoclonal antibody (eprtuzumab)), AVASTIN® (bevacizumab) and TARCEVA® (erlotinib), and analogues thereof. For example, and without wishing to be bound by theory, treatment with an anti-cancer antibody or combination of an anti-cancer antibody and a toxin can cause death of cancer cells (eg, tumor cells), thereby enhancing CTLA-4 or PD-L1-mediated immune responses. . In an exemplary embodiment, the treatment of a proliferative disease (eg, a cancerous tumor) can include an anti-cancer antibody in combination with a macrocyclic peptide and an anti-CTLA-4 agent (either in parallel or sequentially or any combination thereof) that enhances the host Anti-tumor immune response.

Tumors evade host immune surveillance through a variety of mechanisms. Inactivation of proteins that are manifested by tumors and immunosuppressive can overcome many of these mechanisms. These mechanisms include, inter alia, TGF-β (Kehrl, J. et al. , J. Exp. Med. , 163: 1037-1050 (1986)), IL-10 (Howard, M. et al., Immunology Today , 13:198). -200 (1992)) and Fas ligand (Hahne, M. et al., Science , 274: 1363-1365 (1996)). In another example, an antibody against each of these entities can be further combined with the combined macrocyclic peptide and anti-CTLA-4 to counteract the effects of the immunosuppressive agent and contribute to the host's anti-tumor immune response.

Other agents useful for activating host immunoreactivity can be further used in combination with the macrocyclic peptides of the invention. Such agents include molecules that activate DC function and antigen presentation on the surface of dendritic cells. Anti-CD40 macrocyclic peptides are effective in replacing T cell helper activity (Ridge, J. et al, Nature, 393: 474-478 (1998)) and can be used in combination with the macrocyclic peptides of the invention, alone or in combination with anti-CTLA-4. (Ito, N. et al., Immunobiology , 201(5): 527-540 (2000)). Activation is directed to T cell costimulatory molecules (such as OX-40) (Weinberg, A. et al, Immunol. , 164: 2160-2169 (2000)), 4-1BB (Melero, I. et al. , Nat. Med. , The macrocyclic peptides of 3: 682-685 (1997) and ICOS (Hutloff, A. et al., Nature , 397: 262-266 (1999)) also increase the degree of T cell activation.

Bone marrow transplantation is currently used to treat a variety of hematopoietic tumors. Although graft versus host disease is the result of this treatment, therapeutic benefit can be obtained from the graft anti-tumor response. The macrocyclic peptides of the invention, alone or in combination with a CTLA-4 blocker, can be used to enhance tumor implantation in donor donors The effectiveness of heterosexual T cells.

There are also several experimental treatment options, including ex vivo activation and expansion of antigen-specific T cells and their subsequent inheritance transfer to recipients in order to target antigen-specific T cells to tumors (Greenberg, R. et al. , Science , 285: 546-551 (1999)). These methods can also be used to activate T cell responses to infectious agents such as CMV. In the presence of the macrocyclic peptides of the invention alone or in combination with an anti-CTLA-4 antagonist, ex vivo activation is expected to increase the frequency and activity of inherited metastatic T cells.

In certain embodiments, the invention provides methods of using an immunostimulatory agent to alter an adverse event associated with treating a proliferative disorder, comprising administering to the individual a combination of a macrocyclic peptide of the invention and a sub-therapeutic dose of an anti-CTLA-4 antibody . For example, the methods of the invention provide a method of reducing immunostimulatory treatment of antibody-induced colitis or diarrhea by administering a non-absorbable steroid to a patient. Since any patient receiving an immunostimulatory therapeutic antibody is at risk of developing colitis or diarrhea induced by such treatment, this entire patient population is suitable for the therapy according to the methods of the invention. Although steroids have been administered to treat inflammatory bowel disease (IBD) and to prevent worsening of IBD, they have not been used to prevent (reduce morbidity) IBD in patients who have not been diagnosed with IBD. Significant side effects associated with steroids (even non-absorbable steroids) have hampered prophylactic use.

In other embodiments, the macrocyclic peptides of the invention, alone or in combination with a CTLA-4 blocker, can be further used in combination with any non-absorbable steroid. As used herein, a "non-absorbable steroid" is a glucocorticoid that exhibits extensive first-pass metabolism, such that after metabolism in the liver, the bioavailability of the steroid is lower, i.e., less than about 20%. In one embodiment of the invention, the non-absorbable steroid is budesonide. Budesonide is a locally acting glucocorticosteroid that undergoes extensive metabolism (mainly metabolized by the liver) after oral administration. ENTOCORT® EC (Astra-Zeneca) is a pH and time dependent oral budesonide formulation that has been developed to optimize drug delivery to the ileum and the entire colon. ENTOCORT® EC has been approved in the United States for the treatment of mild to moderate Crohn's disease involving the ileum and/or ascending colon. The general oral dose of ENTOCORT® EC for the treatment of Crohn's disease is 6 mg to 9 mg daily. ENTOCORT® EC is released into the intestine prior to absorption and remains in the intestinal mucosa. Once ENTOCORT® EC passes through the intestinal mucosa target tissue, it undergoes extensive metabolism by the cytochrome P450 system in the liver to produce negligible metabolites of glucocorticoid activity. Therefore, bioavailability is low (about 10%). The lower bioavailability of budesonide results in an improved therapeutic ratio compared to other glucocorticoids that are less widely metabolized by the first pass. Budesonide has fewer adverse effects than systemic corticosteroids, including hypothalamic pituitary suppression. However, long-term administration of ENTOCORT® EC produces systemic glucocorticoid effects such as adrenal cortical hyperactivity and adrenal suppression. See Physicians' Desk Reference Supplement , 58th Edition, 608-610 (2004).

In other embodiments, the combination of PD-L1 and CTLA-4 blocker (ie, immunostimulatory therapeutic macrocyclic peptide anti-PD-L1 and anti-CTLA-4) in combination with a non-absorbable steroid may further be combined with a salicylate combination. Salicylates include 5-ASA agents such as: sulfasalazine (AZULFIDINE®, Pharmacia &Upjohn); olsalazine (DIPENTUM®, Pharmacia &UpJohn); balsalazide ( COLAZAL®, Salix Pharmaceuticals, Inc.); and mesalamine (ASACOL®, Procter & Gamble Pharmaceuticals; PENTASA®, Shire US; CANASA®, Axcan Scandipharm, Inc.; ROWASA®, Solvay).

Dosage and formulation

Suitable peptides of formula I, or more particularly, macrocyclic peptides described herein, can be administered to a patient as a separate compound and/or in admixture with an acceptable carrier in the form of a pharmaceutical formulation for the treatment of diabetes and other related disorders. Those skilled in the art of treating diabetes are readily able to determine the dosage and route of administration of a compound to a mammal, including a human, in need of such treatment. Routes of administration may include, but are not limited to, oral, intraoral, rectal, transdermal, buccal, intranasal, pulmonary, subcutaneous, intramuscular, intradermal, sublingual, intracolonic, intraocular, intravenous or intestinal. versus. The compounds are formulated according to the route of administration, based on acceptable pharmaceutical practices (Fingl et al, in The Pharmacological Basis of Therapeutics , Chapter 1, page 1 (1975); Remington's Pharmaceutical Sciences , 18th Edition, Mack Publishing Co., Easton, PA (1990)).

The pharmaceutically acceptable peptide compositions described herein can be administered in a variety of dosage forms, such as lozenges, capsules (each comprising a sustained release or time release formulation), pills, powders, granules, elixirs, in situ coagulation Glues, microspheres, crystal complexes, liposomes, microemulsions, elixirs, suspensions, syrups, aerosol sprays and emulsions. The compositions described herein can also be administered orally, intravenously (intra- or infusion), intraperitoneally, subcutaneously, transdermally or intramuscularly, in all forms using dosage forms well known to those of ordinary skill in the art. The compositions may be administered separately, but are usually administered with a pharmaceutical carrier selected according to the chosen route of administration and standard pharmaceutical practice.

The dosage regimen of the compositions described herein will of course vary depending on known factors, such as the pharmacodynamic profile of the particular agent and its mode of administration and route; the species, age, sex, health, medical condition and weight of the recipient; symptoms The nature and extent of; the type of concurrent therapy; the frequency of treatment; the route of administration; the kidney and liver function of the patient; and the desired effect. The physician or veterinarian can determine and specify the effective amount of the drug necessary to prevent, combat or arrest the progression of the disease.

According to the general rule, the daily oral dose of the active ingredient when used according to the specified effect is in the range of about 0.001 mg to 1000 mg per kg body weight, preferably between about 0.01 mg and 100 mg per kg body weight per day, and preferably at about daily. Between 0.6 and 20 mg/kg. During a constant rate infusion, the active ingredient will have an intravenous daily dose of from 0.001 ng/min to 100.0 ng/min per kilogram of body weight depending on the intended effect. This constant rate intravenous infusion is preferably administered at a rate of from 0.01 ng/min to 50 ng/min per kg of body weight and is optimally administered at a rate of from 0.01 ng/min to 10.0 mg/min per kg of body weight. The compositions described herein can be administered in a single daily dose, or the total daily dose can be administered in divided doses of two, three or four times daily. this The compositions described herein can also be administered by a tank-type formulation that allows for sustained release of the drug over a period of days/weeks/months as needed.

The compositions described herein can be administered in intranasal form, via topical use as a suitable intranasal vehicle, or via a transdermal route using a transdermal patch. When administered in the form of a transdermal delivery system, the dosage administration is of course continuous throughout the dosage regimen, rather than intermittent.

The compositions are typically administered as a mixture with a suitable pharmaceutical diluent, excipient or carrier (herein collectively referred to as a pharmaceutical carrier), which are administered according to the specified mode of administration (i.e., oral lozenges, capsules, elixirs). The aerosol sprays, and syrups produced with or without the propellant are suitably selected and conform to conventional pharmaceutical practices.

For example, when administered orally in the form of a lozenge or capsule, the active pharmaceutical ingredient can be combined with a pharmaceutically acceptable oral non-toxic inert carrier such as, but not limited to, lactose, starch, sucrose, glucose, Cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol and sorbitol; when administered orally in liquid form, the oral drug component can be combined with any pharmaceutically acceptable oral non-toxic inert carrier Combinations of agents such as, but not limited to, ethanol, glycerol, and water. In addition, suitable binders, lubricants, disintegrants, and coloring agents may also be incorporated into the mixture when needed or necessary. Suitable binders include, but are not limited to, starch, gelatin, natural sugars such as, but not limited to, glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate. ), carboxymethyl cellulose, polyethylene glycol and wax. Lubricants used in such dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride. Disintegrators include, but are not limited to, starch, methylcellulose, agar, bentonite, and merlot.

The compositions described herein can also be administered in the form of a mixed microcell or liposome delivery system, such as a single layer of small vesicles, a single layer of large vesicles, and multiple layers of vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, octadecylamine or phospholipid choline. A penetration enhancer can be added to enhance drug absorption.

Since prodrugs are known to enhance a variety of desirable qualities of the drug (i.e., solubility, bioavailability, manufacturability, etc.), the compounds described herein can be delivered in prodrug form. Accordingly, the subject matter described herein is intended to encompass the prodrugs of the compounds claimed herein, methods of delivering the same, and compositions containing the same.

The compositions described herein can also be coupled to a soluble polymer as a targeted pharmaceutical carrier. Such polymers may include polyvinylpyrrolidone, piper copolymer, polyhydroxypropyl-methacrylamide-phenol, polyhydroxyethylaspartamide phenol, or polyoxyethylene-polylysine ( Substituted by palmitoyl residues). In addition, the compositions described herein may be combined with a class of biodegradable polymers suitable for achieving controlled release of a drug, such as polylactic acid, polyglycolic acid, copolymers of polylactic acid and polyglycolic acid, poly-ε-caprolactone, poly Crosslinked or amphoteric block copolymers of hydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates, and hydrogels.

Dosage forms (pharmaceutical compositions) suitable for administration may contain from about 0.01 mg to about 500 mg of active ingredient per dosage unit. In such pharmaceutical compositions, the active ingredient will generally be present in an amount of from about 0.5% to about 95% by weight based on the total weight of the composition.

Gelatin capsules may contain the active ingredient and powdery carriers such as lactose, starch, cellulose derivatives, magnesium stearate and stearic acid. Similar diluents can be used to prepare compressed tablets. Both lozenges and capsules can be prepared as sustained release products to provide continuous release of the drug over a period of hours. The compressed lozenges may be coated with a sugar coating or coated with a film coating to mask any unpleasant taste and protect the tablet from contact with air, or may be coated with a casing to selectively disintegrate in the gastrointestinal tract.

Oral liquid dosage forms can contain coloring and flavoring to enhance patient acceptance.

In general, water, suitable oils, physiological saline, dextrose (glucose) aqueous solutions and related sugar solutions and glycols such as propylene glycol or polyethylene glycol are carriers suitable for parenteral solutions. The solution for parenteral administration preferably contains a water-soluble salt of the active ingredient, a suitable stabilizer, and, if necessary, a buffer substance. Antioxidants, such as sodium bisulfite, sodium sulfite or ascorbic acid, alone or in combination, are suitable stabilizers. Citric acid and its salts are also used. Sodium EDTA. Alternatively, the parenteral solution may contain a preservative such as benzalkonium chloride, methyl or propylparaben, and chlorobutanol.

Suitable pharmaceutical carriers are described in Remington: The Science and Practice of Pharmacy , 19th edition, Mack Publishing Company (1995) (standard reference textbook in this field).

Representative pharmaceutical dosage forms suitable for administration of the compounds described herein can be illustrated as follows:

capsule

A large number of unit capsules can be prepared by filling 100 mg of the powdered active ingredient, 150 mg of lactose, 50 mg of cellulose and 6 mg of magnesium stearate into standard two-piece hard gelatin capsules.

Soft gelatin capsule

A mixture of the active ingredient in a digestible oil such as soybean oil, cottonseed oil or olive oil can be prepared and injected into the gelatin by means of a positive displacement pump to form a soft gelatin capsule containing 100 mg of the active ingredient. The capsule should be washed and dried.

Lozenge

Tablets can be prepared by conventional procedures such that the unit dosage is, for example, 100 mg of active ingredient, 0.2 mg of colloidal cerium oxide, 5 mg of magnesium stearate, 275 mg of microcrystalline cellulose, 11 mg of starch, and 98.8 mg of lactose. Appropriate coatings may be applied to enhance palatability or delay absorption.

injection

Injectable formulations of the peptide compositions described herein may or may not require the use of excipients, such as those already approved by regulatory agencies. Such excipients include, but are not limited to, solvents and cosolvents, solubilizers, emulsifying or thickening agents, chelating agents, antioxidants and reducing agents, antimicrobial preservatives, buffers and pH adjusters, accumulating agents, Protective agent and tension regulator and special additives. Injectable formulations must be sterile, pyrogen free, and in the case of solutions, free of particulate matter.

Parenteral compositions suitable for injection administration can be prepared by stirring, for example, 1.5% by weight of the active ingredient in a pharmaceutically acceptable buffer which may or may not contain a cosolvent or other excipient. The solution should be isotonic and sterilized using sodium chloride.

suspension

Aqueous suspensions may be prepared according to oral and/or parenteral administration such that, for example, 100 mg of finely divided active ingredient per 5 mL, 20 mg of sodium carboxymethylcellulose, 5 mg of sodium benzoate, 1.0 g of sorbitol solution (USP) , and 0.025mL vanilla extract or other palatable flavoring.

Biodegradable microparticles

A sustained release parenteral composition suitable for injection administration can be prepared, for example, by dissolving a suitable biodegradable polymer in a solvent, adding the active agent to be incorporated to the polymer solution, and from the matrix. The solvent is removed to form a polymer matrix in which the active agent is dispersed throughout the matrix.

Peptide synthesis

The macrocyclic peptides of the invention can be produced by methods known in the art, such as chemical synthesis, recombinant synthesis in cell-free systems, recombinant synthesis in cells, or isolation from biological sources. The chemical synthesis of the macrocyclic peptides of the invention can be carried out using a variety of methods recognized in the art, including stepwise solid phase synthesis, semi-synthesis via conformational re-ligation of peptide fragments, and the selection or synthesis of peptide segments. Enzymatic linkage, and chemical linkage. A preferred method of synthesizing the macrocyclic peptides and analogs thereof described herein is chemical synthesis using various solid phase techniques, such as those described in the literature: Chan, WC et al., Fmoc Solid Phase Synthesis , Oxford University Press , Oxford (2000); Barany, G. et al, The Peptides: Analysis, Synthesis, Biology , Volume 2: "Special Methods in Peptide Synthesis, Part A", Vol. 3-284, Gross, E., et al. , Academic Press, New York (1980); and Stewart, JM et al, Solid-Phase Peptide Synthesis , 2nd ed., Pierce Chemical Co., Rockford, IL (1984). A preferred strategy is based on the combination of a Fmoc(9-fluorenylmethylmethoxycarbonyl) group that temporarily protects the alpha-amino group from a third butyl group that temporarily protects the amino acid side chain. (See, for example, Atherton, E., et al., "The Fluorenylmethoxycarbonyl Amino Protecting Group", in The Peptides: Analysis, Synthesis, Biology , Vol. 9: "Special Methods in Peptide Synthesis, Part C", pp. 1-38, Undenfriend , S. et al., eds., Academic Press, San Diego (1987).

Starting from the C-terminus of the peptide, the peptide can be synthesized in a stepwise manner on an insoluble polymeric carrier (also known as "resin"). The C-terminal amino acid starting to synthesize is a peptide attached to the resin via the formation of a guanamine bond or an ester bond. This results in the final release of the peptide in the form of a C-terminal guanamine or a carboxylic acid, respectively.

The C-terminal amino acids used in the synthesis and the alpha-amine groups and side chain functional groups (if present) of all other amino acids must be differentially protected to selectively remove alpha-amine protection during synthesis. base. The coupling of the amino acid is carried out by reacting its carboxyl group with the active ester form and reacting it with the unblocked a-amine group attached to the N-terminal amino acid of the resin. The α-amino protecting group is removed and coupled in sequence until the entire peptide sequence is assembled. Next, in the case of concomitant removal of the protecting group of the side chain functional group, the peptide is usually released from the resin in the presence of a suitable scavenger to limit side reactions. The resulting peptide was finally purified by reverse phase HPLC.

The precursor peptidyl resin required for the synthesis of the final peptide was a commercially available crosslinked polystyrene polymer resin (Novabiochem, San Diego, CA; Applied Biosystems, Foster City, CA). For the C-terminal carboxamide, the preferred solid carrier is: 4-(2',4'-dimethoxyphenyl-Fmoc-aminomethyl)-phenoxyethyl-p-methyl Benzylamine resin (Rink guanamine MBHA resin); 9-Fmoc-amino group- -3-yloxy-Merrifield resin (Sieber decylamine resin); 4-(9-Fmoc)aminomethyl-3,5-dimethoxyphenoxy)pentanyl-aminomethyl-Merrifield Resin (PAL resin). Coupling of the first amino acid and the subsequent amino acid can be performed using HOBt, 6 produced by DIC/HOBt, HBTU/HOBt, BOP, PyBOP or by DIC/6-Cl-HOBt, HCTU, DIC/HOAt or HATU, respectively. -Cl-HOBt or HOAt active ester is completed. For protected peptide fragments, preferred solid carriers are: 2-chlorotrityl chloride resin and 9-Fmoc-amino group- -3-yloxy-Merrifield resin (Sieber amide resin). Loading of the first amino acid onto the 2-chlorotrityl chloride resin is preferably accomplished by reacting the Fmoc protected amino acid with the resin in dichloromethane and DIEA. If necessary, a small amount of DMF may be added to promote dissolution of the amino acid.

Synthesis of the peptide analogs described herein can be performed using a single channel or multi-channel peptide synthesizer, such as a CEM Liberty microwave synthesizer, or a Protein Technologies, Inc. Prelude (6 channel) or Symphony (12 channel) synthesizer.

Suitable Fmoc amino acid derivatives are shown below.

Examples of orthogonally protected amino acids for solid phase synthesis

Peptide-based resin of the respective peptide and the precursor may be split using any standard deprotection procedure (see, e.g. King, DS et al., Int.J.Peptide Protein Res, 36:. 255-266 (1990)). The desired method is to use TFA as a scavenger in the presence of water and TIS. Typically, the peptidyl resin is stirred in TFA/water/TIS (94:3:3, v:v:v; 1 mL/100 mg peptidyl resin) at room temperature for 2 to 6 hours. The waste resin is then filtered off and the TFA solution is concentrated or dried under reduced pressure. The crude peptide obtained was precipitated and washed with Et ₂ O or directly redissolved in DMSO or 50% aqueous acetic acid for purification by preparative HPLC.

Peptides having the desired purity can be obtained by purification using preparative HPLC (for example, on a Waters Model 4000 or Shimadzu LC-8A liquid chromatograph). The crude peptide solution was injected into a YMC S5 ODS (20X 100 mm) column and eluted with a linear gradient of MeCN/water (all buffered with 0.1% TFA) using a flow rate of 14 to 20 mL/min, wherein the effluent was UV based on 220 nm. Absorbance monitoring. The structure of the purified peptide can be confirmed by electrospray MS analysis.

A list of non-naturally occurring amino acids referred to herein is provided below.

The following abbreviations are used in the examples and elsewhere:

Ph = phenyl

Bn = benzyl

i-Bu=isobutyl

i-Pr=isopropyl

Me=methyl

Et=ethyl

Pr = n-propyl

Bu=n-butyl

t-Bu=Third butyl

Trt = trityl

TMS = trimethyl decyl

TIS=triisopropyldecane

Et ₂ O=ether

HOAc or AcOH=acetic acid

MeCN or AcCN=acetonitrile

DMF=N,N-dimethylformamide

EtOAc = ethyl acetate

THF = tetrahydrofuran

TFA = trifluoroacetic acid

TFE=α,α,α-trifluoroethanol

Et ₂ NH=diethylamine

NMM=N-methylmorpholine

NMP=N-methylpyrrolidone

DCM = dichloromethane

TEA = triethylamine

Min.=minute

h or hr = hour

L = liter

mL or ml=ml

μL=microliter

g=g

Mg=mg

Mol=mole

Mmmol=mole

Meq=milli equivalent

Rt or RT = room temperature

Sat or sat'd=saturated

Aq.=aqueous

Mp=melting point

BOP reagent = benzotriazol-1-yloxy-paraxyl-dimethylamino ruthenium (Castro's reagent)

PyBOP reagent = benzotriazol-1-yloxy-tripyrrolidinium hexafluorophosphate

HBTU=2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyl hexafluorophosphate

HATU=O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl hexafluorophosphate

HCTU=2-(6-chloro-1H-benzotriazol-1-yl)-1,1,3,3-tetramethyl hexafluorophosphate

T3P=2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphonium-2,4,6-trioxide

DMAP = 4-(dimethylamino)pyridine

DIEA=diisopropylethylamine

Fmoc or FMOC=mercaptomethoxycarbonyl

Boc or BOC = third butoxycarbonyl

HOBT or HOBT ̇H ₂ O=hydrated 1-hydroxybenzotriazole

Cl-HOBt=6-chloro-benzotriazole

HOAT=1-hydroxy-7-azabenzotriazole

HPLC=High Performance Liquid Chromatography

LC/MS=High Performance Liquid Chromatography/Mass Spectrometry

MS or Mass Spec = Mass Spectrometry

NMR=NMR

Sc or SC = subcutaneous

IP or ip = intraperitoneal

Instance Preparation of Example 1240

Example 1240 was prepared on Rink resin following the general synthetic sequence described for the preparation of Example 0001. The general synthetic sequence consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ""Symphony Method B: Secondary Amine Coupling Procedure ", "Manual Coupling Procedure A", "Symphony Method B: Final Ending Procedure ", " Total Removal Protection Method F " and " Circularization Method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 10-65% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.45 min; ESI-MS (+) m/z 955.0 (M+2H).

Analytical LCMS condition E: retention time = 1.39min; ESI-MS (+ ) m / z 954.7 (M + 2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 1241

Example 1241 was prepared on Rink resin following the general synthetic sequence described for the preparation of Example 0001. The general synthetic sequence consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ""Symphony Method B: Secondary Amine Coupling Procedure ", "Manual Coupling Procedure A", "Symphony Method B: Final Ending Procedure ", " Total Removal Protection Method F " and " Circularization Method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 10-65% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.3 mg and the purity was estimated to be 92% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.37 min; ESI-MS (+) m/z 947.8 (M+2H).

Analytical LCMS Condition E: Retention time = 1.30 min; ESI-MS (+) m/z 948.0 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 1244

Example 1244 was prepared on Rink resin following the general synthetic sequence described for the preparation of Example 0001. The general synthetic sequence consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ""Symphony Method B: Secondary Amine Coupling Procedure ", "Manual Coupling Procedure A", "Symphony Method B: Final Ending Procedure ", " Total Removal Protection Method F " and " Circularization Method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 10-65% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.9 mg and the purity was estimated to be 97% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.49 min; ESI-MS (+) m/z 942.0 (M+2H).

Analytical LCMS Condition E: Retention time = 1.56 min; ESI-MS (+) m/z 942.0 (M+2H).

ESI-HRMS(+) m/z:; Calculated: 941.4771 (M+2H) ; calc .: 941.4757 (M+2H)

Preparation of Example 1245

Example 1245 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Prelude Method C: Resin Swelling Procedure ", "Prelude Method C: Single Coupling Procedure ", "Prelude Method" C: secondary amine coupling procedure , "Prelude method C: final washing procedure ", chloroacetic acid coupling procedure B , " total removal of protecting group method C " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 10-65% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 1.4 mg and the purity was estimated to be 90% according to LCMS analysis.

Analytical LCMS conditions D: residence time = 1.48 min; ESI-MS (+) m/z </ RTI>

Analytical LCMS Condition E: Retention time = 1.42 min; ESI-MS (+) m/z: 907.3 (M+2H).

Preparation of Example 1246

Example 1246 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Prelude Method C: Resin Swelling Procedure ", "Prelude Method C: Single Coupling Procedure ", "Prelude Method" C: secondary amine coupling procedure , "Prelude method C: final washing procedure ", chloroacetic acid coupling procedure B , " total removal of protecting group method C " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 15-65% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.5 mg and the purity was estimated to be 94% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.57 min; ESI-MS (+) m/z 941.9 (M+2H).

Analytical LCMS Condition E: Retention time = 1.51 min; ESI-MS (+) m/z 942.2 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 941.4771 (M + 2H); Found: 941.4755 (M + 2H).

Preparation of Example 1247

Example 1247 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Prelude Method C: Resin Swelling Procedure ", "Prelude Method C: Single Coupling Procedure ", "Prelude Method" C: secondary amine coupling procedure , "Prelude method C: final washing procedure ", chloroacetic acid coupling procedure B , " total removal of protecting group method C " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 10-65% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 1.1 mg and the purity was estimated to be 94% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.49 min; ESI-MS (+) m/z 921.1 (M+2H).

Analytical LCMS Condition E: Retention time = 1.42 min; ESI-MS (+) m/z 921.4 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 920.9456 (M + 2H); Found: 920.9436 (M + 2H).

Preparation of Example 1248

Example 1248 was prepared following the general synthetic sequence for the preparation of examples of the 0001, the general synthetic sequence consisting of the following general procedure: "Prelude Method:" Prelude single coupling procedure Method C ":" Prelude Method C resin was swollen program ", C: secondary amine coupling procedure , "Prelude method C: final washing procedure ", chloroacetic acid coupling procedure B , " total removal of protecting group method C " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 10-65% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.7 mg and the purity was estimated to be 94% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.45 min; ESI-MS (+) m/z 914.3 (M+2H).

Analytical LCMS Condition E: Retention time = 1.40 min; ESI-MS (+) m/z 914.1 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 913.9378 (M + 2H); Found: 913.9372 (M + 2H).

Preparation of Example 1250

Example 1250 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified by preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm; 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.4 mg and the purity was estimated to be 96% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.46 min.; ESI-MS (+) m/z 962.7 (M+2H).

Analytical LCMS conditions I: residence time = 2.10 min.; ESI-MS (+) m/z 962.5 (M+2H).

ESI-HRMS(+) m/z:; Calculated: 962.4460 (M+2H) ; Found : 962.4459 (M+2H)

Preparation of Example 1251

Example 1251 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method D " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: Column: XBridge C18, 19×200 mm; 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-85% B for 30 minutes, It was then held at 100% B for 5 minutes; flow rate: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.68 min.; ESI-MS (+) m/z 926.4 (M+2H).

Analytical LCMS conditions I: residence time = 3.10 min.; ESI-MS (+) m/z 926.4 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 925.9560 (M + 2H); Found: 925.9548 (M + 2H).

Preparation of Example 1252

Example 1252 was prepared on Rink resin following the general synthetic sequence described for the preparation of Example 0002. The general synthetic sequence consisted of the following general procedure: "CEM Method A: Resin Swelling Procedure", "CEM Method A: Single Coupling Procedure" , "CEM Method A: Double Coupling Procedure", "Chloroethyl Chloride Chloride Coupling Procedure A", "Total Removal of Protective Group Method F" and "Circulation Method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19×mm; 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; Mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: residence time = 1.48 min.; ESI-MS (+) m/z 963.1 (M+2H).

Analytical LCMS condition I: retention time = 2.08min; ESI-MS (+ ) m / z 963.5 (M + 2H)..

ESI-HRMS (+) m / z :; Calcd: 962.9380 (M + 2H); Found: 962.9370 (M + 2H).

Preparation of Example 1255

Example 1255 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method D " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B duration 30 Minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.58 min.; ESI-MS (+) m/z 933.6 (M+2H).

Analytical LCMS conditions I: residence time = 2.25 min.; ESI-MS (+) m/z 933.5 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 933.4419 (M + 2H); Found: 933.4432 (M + 2H).

Preparation of Example 1256

Example 1256 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method D " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.2 mg, and the root According to LCMS analysis, the purity is estimated to be 100%.

Analytical LCMS conditions H: retention time = 1.37 min.; ESI-MS (+) m/z 963.6 (M+2H).

Analytical LCMS Condition I: Retention time = 1.98 min.; ESI-MS (+) m/z 963.6 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 963.4300 (M + 2H); Found: 963.4295 (M + 2H).

Preparation of Example 1257

Example 1257 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method D " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.60 min.; ESI-MS (+) m/z s.

Analytical LCMS conditions I: residence time = 3.12 min.; ESI-MS (+) m/z s.

ESI-HRMS (+) m / z :; Calcd: 908.4298 (M + 2H); Found: 908.4283 (M + 2H).

Preparation of Example 1258

Was prepared following the general synthetic sequence for the preparation of Example 0001 according to Example of 1258, the general synthetic sequence composed by the following general procedure: "Symphony Method B: resin swollen program", "Symphony Method B: Standard coupling procedures", "method Symphony B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method D " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm; 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. Combine the fractions containing the desired product and evaporate by centrifugation Dry it. The yield of the product was 4.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.61 min.; ESI-MS (+) m/z 909.0 (M+2H).

Analytical LCMS conditions I: retention time = 2.66 min.; ESI-MS (+) m/z 909.6 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 908.9218 (M + 2H); Found: 908.9206 (M + 2H).

Preparation of Example 1259

Example 1259 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method D " and " cyclization method D ".

The crude material was purified by preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm; 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. Substance through the use of the following conditions Prepared LC/MS for further purification: column: XBridge C18, 19 × 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95:5 acetonitrile: water + 0.1 % trifluoroacetic acid; gradient: 10-50% B for 30 minutes followed by 100 minutes at 100% B; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.6 mg and its purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.49 min.; ESI-MS (+) m/z 976.7 (M+2H).

Analytical LCMS conditions I: retention time = 2.15 min.; ESI-MS (+) m/z 976.7 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 976.4798 (M + 2H); Found: 976.4781 (M + 2H).

Preparation of Example 1260

Example 1260 was prepared on Rink resin following the general synthetic sequence described for the preparation of Example 0002. The general synthetic sequence consisted of the following general procedure: "CEM Method A: Resin Swelling Procedure", "CEM Method A: Single Coupling Procedure" , "CEM Method A: Double Coupling Procedure", "Chloroethyl Chloride Chloride Coupling Procedure A", "Total Removal of Protective Group Method F" and "Circulation Method C".

The crude material was purified via preparative LC/MS using the following conditions: Column: XBridge C18, 19×mm; 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, It was then held at 100% B for 5 minutes; flow rate: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 35.2 mg and the purity was estimated to be 98% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.51 min.; ESI-MS (+) m/z 963.2 (M+2H).

Analytical LCMS conditions I: retention time = 2.05 min.; ESI-MS (+) m/z 962.9 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 962.9380 (M + 2H); Found: 962.9367 (M + 2H).

Preparation of Example 1261

Example 1261 was prepared on Rink resin following the general synthetic sequence described for the preparation of Example 0002, which consisted of the following general procedure: "CEM Method A: Resin Swelling Procedure", "CEM Method A: Single Coupling Procedure" , "CEM Method A: Double Coupling Procedure", "Chloroethyl Chloride Chloride Coupling Procedure A", "Total Removal of Protective Group Method F" and "Circulation Method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19×mm; 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; Mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.2 mg and the purity was estimated to be 96% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.55 min .; ESI-MS (+) m/z 934.1 (M+2H).

Analytical LCMS conditions I: residence time = 3.02 min.; ESI-MS (+) m/z 934.1 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 933.9352 (M + 2H); Found: 933.9344 (M + 2H).

Preparation of Example 1262

Example 1262 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method D " and " cyclization method D ".

The crude material was purified by preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm; 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. Consolidation contains what you want The product was dissolved and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 25-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.65 min.; ESI-MS (+) m/z 944.8 (M+2H).

Analytical LCMS conditions I: retention time = 2.59 min.; ESI-MS (+) m/z 945.0 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 944.4404 (M + 2H); Found: 944.4388 (M + 2H).

Preparation of Example 1272

Example 1272 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Prelude Method C: Resin Swelling Procedure ", "Prelude Method C: Single Coupling Procedure ", "Prelude Method" C: secondary amine coupling procedure , "Prelude method C: final washing procedure ", chloroacetic acid coupling procedure B , " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified by preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm; 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.6 mg and the purity was estimated to be 98% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.65 min.; ESI-MS (+) m/z 930.4 (M+2H).

Analytical LCMS conditions I: retention time = 3.08 min.; ESI-MS (+) m/z 930.4 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 929.9453 (M + 2H); Found: 929.9429 (M + 2H).

Preparation of Example 1273

Example 1273 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Prelude Method C: Resin Swelling Procedure ", "Prelude Method C: Single Coupling Procedure ", "Prelude Method" C: secondary amine coupling procedure , "Prelude method C: final washing procedure ", chloroacetic acid coupling procedure B , " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.66 min.; ESI-MS (+) m/z 954.8 (M+2H).

Analytical LCMS conditions I: residence time = 3.14 min.; ESI-MS (+) m/z 954.8 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 954.4667 (M + 2H); Found: 954.4644 (M + 2H).

Preparation of Example 1275

Example 1275 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 0.4 mg and the purity was estimated to be 97% according to LCMS analysis.

Analytical LCMS conditions H: residence time = 1.48 min.; ESI-MS (+) m/z 934.4 (M+2H).

Analytical LCMS conditions I: retention time = 2.97 min .; ESI-MS (+) m/z 934.4 (M+2H).

Preparation of Example 1276

Example 1276 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.6 mg and the purity was estimated to be 97% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.54 min.; ESI-MS (+) m/z 933.7 (M+2H).

Analytical LCMS conditions I: retention time = 3.02 min.; ESI-MS (+) m/z 933.4 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 932.9512 (M + 2H); Found: 932.9524 (M + 2H).

Preparation of Example 1277

Example 1277 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B lasts 30 minutes, then remains at 100% B 5 Minute; flow rate: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.61 min.; ESI-MS (+) m/z 933.9 (M+2H).

Analytical LCMS conditions I: residence time = 3.07 min.; ESI-MS (+) m/z 933.9 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 933.4432 (M + 2H); Found: 933.4416 (M + 2H).

Preparation of Example 1278

Example 1278 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.3 mg and the purity was estimated to be 99% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.52 min.; ESI-MS (+) m/z 920.3 (M+2H).

Analytical LCMS conditions I: retention time = 2.97 min.; ESI-MS (+) m/z 920.4 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 919.9434 (M + 2H); Found: 919.9422 (M + 2H).

Preparation of Example 1279

Example 1279 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.1 mg and the purity was estimated to be 99% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.55 min .; ESI-MS (+) m/z 921.0 (M+2H).

Analytical LCMS conditions I: retention time = 2.99 min.; ESI-MS (+) m/z 920.9 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 920.4354 (M + 2H); Found: 920.4340 (M + 2H).

Preparation of Example 1280

Example 1280 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.0 mg and the purity was estimated to be 97% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.48 min.; ESI-MS (+) m/z 929.0 (M+2H).

Analytical LCMS conditions I: residence time = 2.93 min.; ESI-MS (+) m/z 928.9 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 928.4329 (M + 2H); Found: 928.4324 (M + 2H).

Preparation of Example 1281

Example 1281 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Custom Amino Acid Coupling Procedure " , "Symphony Method B: Standard Coupling Program ", "Symphony Method B: Secondary Amine Coupling Program ", "Symphony Method B: Final Ending Procedure ", " Total Removal of Protective Base Method F ", and " Circularization Method D " .

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.9 mg and its purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: residence time = 1.81 min.; ES "-MS (+) m/z 923.7 (M+2H).

Analytical LCMS conditions I: retention time = 2.63 min.; ESI-MS (+) m/z 924.1 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 922.9431 (M + 2H); Found: 922.9422 (M + 2H).

Preparation of Example 1282

Example 1282 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified by preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm; 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.6 mg and the purity was estimated to be 95% according to LCMS analysis.

Analytical LCMS conditions H: residence time = 1.51 min.; ESI-MS (+) m/z 964.3 (M+2H).

Analytical LCMS conditions I: retention time = 2.27 min.; ESI-MS (+) m/z 964.7 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 963.4356 (M + 2H); Found: 963.4356 (M + 2H).

Preparation of Example 1283

Example 1283 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified by preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm; 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 8 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.32 min.; ESI-MS (+) m/z 963.7 (M+2H).

Analytical LCMS Condition I: Retention time = 2.67 min.; ESI-MS (+) m/z 963.8 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 963.9276 (M + 2H); Found: 963.9270 (M + 2H).

Preparation of Example 1285

Example 1285 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , " manual coupling procedure A ", "Symphony method B: final termination procedure ", " total removal protection method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.4 mg and the purity was estimated to be 95% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.86 min.; ESI-MS (+) m/z 928.5 (M+2H).

ESI-HRMS(+) m/z:; calcd.: 927.4432 (M+2H) ;

Preparation of Example 1289

Example 1289 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: custom amino acid coupling procedure ", "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D " .

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.7 mg and the purity was estimated to be 99% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.74 min.; ESI-MS (+) m/z 926.4 (M+2H).

Analytical LCMS conditions I: residence time = 3.32 min.; ESI-MS (+) m/z 926.2 (M+2H).

ESI-HRMS (+) m / z :; Calcd: 925.9378 (M + 2H); Found: 925.9350 (M + 2H).

Preparation of Example 1290

Example 1290 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: water + 0.1% trifluoroacetic acid; mobile phase B: 95:5 acetonitrile: water +0.1% trifluoroacetic acid; gradient: 5-45% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.7 mg and the purity was estimated to be 94% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.24 min.; ESI-MS (+) m/z 967.3 (M+2H).

Analytical LCMS conditions I: retention time = 2.66 min.; ESI-MS (+) m/z 967.4 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 966.9273 (M + 2H); Found: 966.9273 (M + 2H).

Preparation of Example 1291

Example 1291 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-100% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.1 mg and the purity was estimated to be 93% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.39 min.; ESI-MS (+) m/z 944.1 (M+2H).

Analytical LCMS conditions I: residence time = 2.99 min.; ESI-MS (+) m/z 943.8 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 943.4745 (M + 2H); Found: 943.4714 (M + 2H).

Preparation of Example 1292

Example 1292 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 1.6 mg and its purity was estimated according to LCMS analysis. It is 100%.

Analytical LCMS conditions H: residence time = 1.20 min.; ESI-MS (+) m/z 963.3 (M+2H).

Analytical LCMS conditions I: residence time = 2.52 min.; ESI-MS (+) m/z 963.5 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 962.9380 (M + 2H); Found: 962.9357 (M + 2H).

Preparation of Example 1293

Example 1293 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.1 mg and the purity was estimated to be 98% according to LCMS analysis.

Analytical LCMS conditions H: residence time = 1.48 min.; ESI-MS (+) m/z 983.9 (M+2H).

Analytical LCMS conditions I: retention time = 2.12 min.; ESI-MS (+) m/z 984.1 (M+2H).

Preparation of Example 1294

Example 1294 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.1 mg and its purity was estimated according to LCMS analysis. It is 94%.

Analytical LCMS conditions H: retention time = 1.22 min.; ESI-MS (+) m/z 978.8 (M+2H).

Analytical LCMS conditions I: retention time = 2.56 min.; ESI-MS (+) m/z 979.0 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 977.9433 (M + 2H); Found: 977.9417 (M + 2H).

Preparation of Example 1295

Example 1295 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile :water +10 mM ammonium acetate; gradient: 5-45% B for 20 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.38 min.; ESI-MS (+) m/z 951.6 (M+2H).

Analytical LCMS conditions I: retention time = 2.96 min.; ESI-MS (+) m/z 952.0 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 951.4538 (M + 2H); Found: 951.4508 (M + 2H).

Preparation of Example 1296

Example 1296 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B duration 30 Minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 1.4 mg and the purity was estimated to be 97% according to LCMS analysis.

Analytical LCMS conditions H: residence time = 1.35 min.; ESI-MS (+) m/z 1005.2 (M+2H).

Analytical LCMS conditions I: residence time = 2.75 min.; ESI-MS (+) m/z 1005.3 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 1004.5111 (M + 2H); Found: 1004.5078 (M + 2H).

Preparation of Example 1297

Example 1297 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: custom amino acid coupling procedure ", "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D " .

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.58 min.; ESI-MS (+) m/z 957.5 (M+2H).

Analytical LCMS conditions I: residence time = 3.18 min.; ESI-MS (+) m/z 957.5 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 956.9826 (M + 2H); Found: 956.9896 (M + 2H).

Preparation of Example 1298

Example 1298 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: custom amino acid coupling procedure ", "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D " .

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 20 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.33 min.; ESI-MS (+) m/z 977.3 (M+2H).

Analytical LCMS conditions I: retention time = 2.71 min.; ESI-MS (+) m/z 977.3 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 976.4672 (M + 2H); Found: 976.4644 (M + 2H).

Preparation of Example 1299

An example 1299 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: custom amino acid coupling procedure ", "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D " .

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-85% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.3 mg and the purity was estimated to be 92% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.40 min.; ESI-MS (+) m/z 965.7 (M+2H).

Analytical LCMS conditions I: residence time = 2.79 min.; ESI-MS (+) m/z 965.3 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 964.9718 (M + 2H); Found: 964.9680 (M + 2H).

Preparation of Example 1300

Example 1300 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: custom amino acid coupling procedure ", "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D " .

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.3 mg and the purity was estimated to be 97% according to LCMS analysis.

Analytical LCMS conditions I: residence time = 2.65 min.; ESI-MS (+) m/z 944.8 (M+2H).

Preparation of Example 1301

Example 1301 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: custom amino acid coupling procedure ", "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D " .

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.1 mg and the purity was estimated to be 97% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.33 min.; ESI-MS (+) m/z 977.3 (M+2H).

Analytical LCMS conditions I: retention time = 2.71 min.; ESI-MS (+) m/z 977.3 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 976.4672 (M + 2H); Found: 976.4645 (M + 2H).

Preparation of Example 1302

Example 1302 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: custom amino acid coupling procedure ", "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D " .

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. Combine the fractions containing the desired product and evaporate by centrifugation Dry it. The yield of the product was 1.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: residence time = 1.20 min.; ESI-MS (+) m/z 1006.3 (M+2H).

Analytical LCMS conditions I: retention time = 2.47 min.; ESI-MS (+) m/z 1006.3 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 1005.4826 (M + 2H); Found: 1005.4786 (M + 2H).

Preparation of Example 1303

Example 1303 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: custom amino acid coupling procedure ", "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D " .

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. Consolidation contains what you want The product was dissolved and dried by centrifugal evaporation. The yield of the product was 8.0 mg and the purity was estimated to be 93% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.25 min.; ESI-MS (+) m/z 1012.9 (M+2H).

Analytical LCMS Condition I: Retention time = 2.52 min.; ESI-MS (+) m/z 1013.3 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 1011.9984 (M + 2H); Found: 1011.9941 (M + 2H).

Preparation of Example 1304

Example 1304 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: custom amino acid coupling procedure ", "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D " .

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. Consolidation contains what you want The product was dissolved and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 1.1 mg and the purity was estimated to be 95% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.34 min.; ESI-MS (+) m/z 959.0 (M+2H).

Analytical LCMS conditions I: retention time = 2.98 min.; ESI-MS (+) m/z 959.0 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 958.4560 (M + 2H); Found: 958.4542 (M + 2H).

Preparation of Example 1305

Example 1305 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: custom amino acid coupling procedure ", "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D " .

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 0.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.25 min.; ESI-MS (+) m/z 966.4 (M+2H).

Analytical LCMS conditions I: residence time = 2.91 min.; ESI-MS (+) m/z 965.7 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 965.4638 (M + 2H); Found: 965.4619 (M + 2H).

Preparation of Example 1306

Example 1306 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony" Method B: Second Amine Coupling Procedure , "Symphony Method B: Custom Amino Acid Coupling Procedure ", "Symphony Method B: Final Ending Procedure ", " Total Removal of Protective Group Method F ", and " Circulation Method D" "."

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.8 mg and the purity was estimated to be 92% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.57 min.; ESI-MS (+) m/z 951.9 (M+2H).

Analytical LCMS Condition I: Retention time = 3.07 min.; ESI-MS (+) m/z 951.9 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 951.4664 (M + 2H); Found: 951.4633 (M + 2H).

Preparation of Example 1309

Example 1309 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: custom amino acid coupling procedure ", "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D " .

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 0-50% B over 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B : 95:5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 15-55% B for 25 minutes, followed by 55% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation.

The yield of the product was 3.6 mg and the purity was estimated to be 98% according to LCMS analysis.

Analytical LCMS conditions D: residence time = 1.1 min; ESI-MS (+) m/z 925.3 (M+2H).

Analytical LCMS conditions E: residence time = 1.25 min; ESI-MS (+) m/z 925.4 (M+2H).

ESI-HRMS (+) m / z:; Calcd: 924.8985 (M + 2H); Found: 950.8961 (M + 2H).

Preparation of Example 1500

Example 1500 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 0.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: residence time = 1.60 min; ESI-MS (+) m/z 957.00 (M+2H). Analytical LCMS conditions I: residence time = 3.06 min; ESI-MS (+) m/z 957.00 (M+2H).

Preparation of Example 1501

Example 1501 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.4 mg and the purity was estimated to be 97% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.76 min; ESI-MS (+) m/z 928.15 (M+2H).

Analytical LCMS conditions I: residence time = 3.20 min; ESI-MS (+) m/z 928.20 (M+2H).

Preparation of Example 1502

Example 1502 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: residence time = 1.796 min; ESI-MS (+) m/z 941.20 (M+2H).

Analytical LCMS conditions I: residence time = 2.389 min; ESI-MS (+) m/z 940.95 (M+2H).

Preparation of Example 1503

Example 1503 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: residence time = 1.870 min; ESI-MS (+) m/z 948.75 (M+2H).

Analytical LCMS conditions I: residence time = 3.358 min; ESI-MS (+) m/z 948.60 (M+2H).

Preparation of Example 1504

Example 1504 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.4 mg and the purity was estimated to be 98% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.447 min; ESI-MS (+) m/z 970.80 (M+2H).

Analytical LCMS conditions I: residence time = 2.838 min; ESI-MS (+) m/z 970.20 (M+2H).

Preparation of Example 1505

Example 1505 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.9 mg and the purity was estimated to be 98% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.624 min; ESI-MS (+) m/z 970.95 (M+2H).

Analytical LCMS conditions I: residence time = 3.075 min; ESI-MS (+) m/z 970.65 (M+2H).

Preparation of Example 1506

Example 1506 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.7 mg and the purity was estimated to be 98% according to LCMS analysis.

Analytical LCMS conditions H: residence time = 1.566 min; ESI-MS (+) m/z 947.20 (M+2H).

Analytical LCMS conditions I: retention time = 3.069 min; ESI-MS (+) m/z 946.90 (M+2H).

Preparation of Example 1507

Example 1507 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: residence time = 1.761 min; ESI-MS (+) m/z 917.60 (M+2H).

Analytical LCMS conditions I: residence time = 3.321 min; ESI-MS (+) m/z 917.40 (M+2H).

Preparation of Example 1508

Example 1508 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.702 min; ESI-MS (+) m/z 942.90 (M+2H).

Analytical LCMS conditions I: residence time = 3.245 min; ESI-MS (+) m/z 942.95 (M+2H).

Preparation of Example 1509

Example 1509 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.9 mg and the purity was estimated to be 98% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.702 min; ESI-MS (+) m/z 942.90 (M+2H).

Analytical LCMS conditions I: residence time = 3.245 min; ESI-MS (+) m/z 942.95 (M+2H).

Preparation of Example 1510

Example 1510 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.0 mg and the purity was estimated to be 99% according to LCMS analysis.

Analytical LCMS conditions H: residence time = 1.706 min; ESI-MS (+) m/z 917.55 (M+2H).

Analytical LCMS conditions I: retention time = 3.271 min; ESI-MS (+) m/z 917.90 (M+2H).

Preparation of Example 1511

Example 1511 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.819 min; ESI-MS (+) m/z 963.45 (M+2H).

Analytical LCMS conditions I: retention time = 3.377 min; ESI-MS (+) m/z 963.45 (M+2H).

Preparation of Example 1512

Example 1512 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure, "" Symphony method B: final terminated program "," general deprotection method F "and" D cyclization process. "

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.479 min; ESI-MS (+) m/z 972.25 (M+2H).

Analytical LCMS conditions I: residence time = 3.000 min; ESI-MS (+) m/z 972.40 (M+2H).

Preparation of Example 1513

Example 1513 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.8 mg and the purity was estimated to be 98% according to LCMS analysis.

Analytical LCMS conditions H: residence time = 1.539 min; ESI-MS (+) m/z 971.90 (M+2H).

Analytical LCMS conditions I: retention time = 3.071 min; ESI-MS (+) m/z 971.85 (M+2H).

Preparation of Example 1514

Example 1514 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.66 min; ESI-MS (+) m/z 935.5 (M+2H).

Analytical LCMS conditions I: retention time = 2.53 min; ESI-MS (+) m/z 935.7 (M+2H).

Preparation of Example 1515

Example 1515 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.455 min; ESI-MS (+) m/z 964.65 (M+2H).

Analytical LCMS conditions I: residence time = 2.809 min; ESI-MS (+) m/z 964.75 (M+2H).

Preparation of Example 1519

Example 1519 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.551 min; ESI-MS (+) m/z 922.85 (M+2H).

Analytical LCMS conditions I: residence time = 3.012 min; ESI-MS (+) m/z 922.90 (M+2H).

Preparation of Example 1520

Example 1520 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-65% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.30 min; ESI-MS (+) m/z 939.6 (M+2H).

Analytical LCMS Condition E: Retention time = 1.40 min; ESI-MS (+) m/z 939.4 (M+2H).

Preparation of Example 1521

Example 1521 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.9 mg and the purity was estimated to be 97% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.525 min; ESI-MS (+) m/z 938.15 (M+2H).

Analytical LCMS conditions I: retention time = 2.936 min; ESI-MS (+) m/z 938.05 (M+2H).

Preparation of Example 1522

Example 1522 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.637 min; ESI-MS (+) m/z 901.95 (M+2H).

Analytical LCMS conditions I: residence time = 3.038 min; ESI-MS (+) m/z 901.75 (M+2H).

Preparation of Example 1523

Example 1523 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.6 mg and the purity was estimated to be 92% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.683 min; ESI-MS (+) m/z 900.80 (M+2H).

Analytical LCMS Condition I: Retention time = 3.098 min; ESI-MS (+) m/z 900.60 (M+2H).

Preparation of Example 1525

Example 1525 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.643 min; ESI-MS (+) m/z 927.20 (M+2H).

Analytical LCMS conditions I: residence time = 2.250 min; ESI-MS (+) m/z 926.85 (M+2H).

Preparation of Example 1526

Example 1526 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 10-65% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.0 mg and the purity was estimated to be 94% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.39 min; ESI-MS (+) m/z 944.3 (M+2H).

Analytical LCMS Condition E: Retention time = 1.44 min; ESI-MS (+) m/z 944.2 (M+2H).

Preparation of Example 1528

Example 1528 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 10-60% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 15-60% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.1 mg and the purity was estimated to be 95% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.43 min; ESI-MS (+) m/z 937.1 (M+2H).

Analytical LCMS conditions E: residence time = 1.41 min; ESI-MS (+) m/z 937.5 (M+2H).

Preparation of Example 1529

Example 1529 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-60% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.1 mg and the purity was estimated to be 91% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.44 min; ESI-MS (+) m/z 927.0 (M+2H).

Analytical LCMS Condition E: Retention time = 1.35 min; ESI-MS (+) m/z 926.8 (M+2H).

Preparation of Example 1530

Example 1530 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: waters xbridgee-18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 0.1% trifluoroacetic acid; mobile phase B: 95:5 methanol: water + 0.1% trifluoroacetic acid; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.9 mg and its purity according to LCMS analysis. The estimate is 100%.

Analytical LCMS conditions H: retention time = 1.57 min; ESI-MS (+) m/z 931.9 (M+2H).

Analytical LCMS conditions I: residence time = 2.70 min; ESI-MS (+) m/z 931.2 (M+2H).

Preparation of Example 1531

Example 1531 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 10-60% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 1.2 mg and the purity was estimated to be 98% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.34 min; ESI-MS (+) m/z 963.0 (M+2H).

Analytical LCMS Condition E: Retention time = 1.35 min; ESI-MS (+) m/z 962.6 (M+2H).

Preparation of Example 1532

Example 1532 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 0.1% trifluoroacetic acid; mobile phase B : 95:5 methanol: water + 0.1% trifluoroacetic acid; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.2 mg and the purity was estimated to be 94% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.48 min; ESI-MS (+) m/z 949.4 (M+2H).

Analytical LCMS conditions I: retention time = 2.57 min; ESI-MS (+) m/z 949.0 (M+2H).

Preparation of Example 1533

Example 1533 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 0.1% trifluoroacetic acid; mobile phase B : 95:5 methanol: water + 0.1% trifluoroacetic acid; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.63 min; ESI-MS (+) m/z 971.6 (M+2H).

Analytical LCMS Condition I: Retention time = 2.87 min; ESI-MS (+) m/z 971.9 (M+2H).

Preparation of Example 1534

Example 1534 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 0-50% B over 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 0-45% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 1.1 mg and its purity according to LCMS analysis. The estimate is 95%.

Analytical LCMS conditions D: retention time = 0.94 min; ESI-MS (+) m/z 922.1 (M+2H).

Analytical LCMS conditions E: residence time = 0.97 min; ESI-MS (+) m/z 921.5 (M+2H).

Preparation of Example 1535

Example 1535 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 10-60% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 B Nitrile: water + 0.1% trifluoroacetic acid; gradient: 5-50% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.32 min; ESI-MS (+) m/z 948.5 (M+2H).

Analytical LCMS Condition E: Retention time = 1.21 min; ESI-MS (+) m/z 948.7 (M+2H).

Preparation of Example 1536

Example 1536 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 10-60% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. Consolidation The fractions of the desired product were dried and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 15-60% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 1.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.32 min; ESI-MS (+) m/z 956.7 (M+2H).

Analytical LCMS Condition E: Retention time = 1.43 min; ESI-MS (+) m/z 956.7 (M+2H).

Preparation of Example 1537

Example 1537 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: Column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 10-60% B for 25 minutes , then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-55% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 1.9 mg and the purity was estimated to be 99% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.42 min; ESI-MS (+) m/z 969.7 (M+2H).

Analytical LCMS Condition E: Retention time = 1.40 min; ESI-MS (+) m/z 969.7 (M+2H).

Preparation of Example 1538

Example 1538 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure, "" Symphony method B: final terminated program "," general protecting group removal method except F "and" D cyclization process. "

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 10-60% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.38 min; ESI-MS (+) m/z 977.6 (M+2H).

Analytical LCMS Condition E: Retention time = 1.31 min; ESI-MS (+) m/z 976.7 (M+2H).

Preparation of Example 1541

Example 1541 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroethyl Acid; mobile phase B: 95:5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.4 mg and the purity was estimated to be 96% according to LCMS analysis.

Analytical LCMS conditions I: residence time = 2.87 min; ESI-MS (+) m/z 960.7 (M+2H).

Preparation of Example 1542

Example 1542 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method B: Resin Swelling Procedure ", "Symphony Method B: Standard Coupling Procedure ", "Symphony Method" B: secondary amine coupling procedure , "Symphony method B: final termination procedure ", " total removal of protecting group method F " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.7 mg and the purity was estimated to be 97% according to LCMS analysis.

Analytical LCMS conditions H: retention time = 1.769 min; ESI-MS (+) m/z 929.30 (M+2H).

Analytical LCMS conditions I: residence time = 3.313 min; ESI-MS (+) m/z 929.30 (M+2H).

Analysis data:

Mass spectrometry: "ESI-MS (+)" indicates electrospray ionization mass spectrometry in positive ion mode; "ESI-MS (-)" indicates electrospray ionization mass spectrometry in negative ion mode; "ESI-HRMS (+)" indicates High-resolution electrospray ionization mass spectrometry in positive ion mode; "ESI-HRMS(-)" represents high-resolution electrospray ionization mass spectrometry in negative ion mode. The detected quality is reported after the " m/z " unit name. Compounds with an accurate mass greater than 1000 are often detected as ions with double or triple charges.

Analytical LCMS Condition A:

Column: Waters BEH C18, 2.1 × 50 mm, 1.7 μm particles; mobile phase A: water + 0.05% TFA; mobile phase B: acetonitrile + 0.05% TFA; temperature: 50 ° C; gradient: 2% B to 98% B duration 2 minutes, followed by 98% B for 0.5 minutes; flow rate: 0.8 mL/min; detection: 220 nm UV.

Analytical LCMS Condition C:

Column: Waters BEH C18, 2.1 x 50 mm, 1.7 μm particles; mobile phase A: water + 0.2% formic acid and 0.01% TFA; mobile phase B: acetonitrile + 0.2% formic acid and 0.01% TFA; temperature: 50 ° C; 2% B to 80% B lasts 2 minutes, 80% B to 98% B lasts 0.1 minutes, then remains at 98% B for 0.5 minutes; flow rate: 0.8 mL/min; detection: 220 nm UV.

Analytical LCMS Condition D:

Column: Waters BEH C18, 2.1×50 mm, 1.7 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; temperature: 50 ° C; Gradient: 0-100% B for 3 minutes, then at 100% B for 0.75 minutes; flow rate: 1.0 mL/min; detection: 220 nm UV.

Analytical LCMS Condition E:

Column: Waters BEH C18, 2.1 × 50 mm, 1.7 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95:5 acetonitrile: water + 0.1% trifluoroacetic acid; : 50 ° C; Gradient: 0-100% B for 3 minutes, followed by 100% B for 0.75 minutes; flow rate: 1.11 mL/min; detection: 220 nm UV.

Analytical LCMS Condition F:

Column: Waters XBridge C18, 2.1×50 mm; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; temperature: 35 ° C; gradient: 0- 100% B lasted 4 minutes, then held at 100% B for 1 minute; flow rate: 4 mL/min; detection: 220 nm UV.

Analytical LCMS Condition G:

Column: Waters BEH C18, 2.0×50 mm, 1.7 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; temperature: 50 ° C; Gradient: 0-100% B for 3 minutes, then at 100% B for 0.5 minutes; flow rate: 0.5 mL/min; detection: 220 nm UV.

Analytical HPLC Condition B:

Column: YMC Pack ODS-AQ 3um 150x4.6mm; mobile phase A: water +0.1% TFA; mobile phase B: acetonitrile + 0.1% TFA; temperature: 40 ° C; gradient: 10% B to 100% duration 10 to 40 Minutes; flow rate: 1 mL/min; detection: 220 nm UV.

General procedure: Prelude Method A:

All operations were performed on a Prelude Peptide Synthesizer (Protein Technologies) according to automated operation. Unless otherwise noted, all procedures were performed in 10 mL or 45 mL polypropylene tubes with bottom frit. The tube is connected to the Prelude Peptide Synthesizer via the bottom and top of the tube. DMF and DCM can be added via the top of the tube, which is equally flushed along the side of the tube. The remaining reagent is added via the bottom of the tube and passed up through the frit to contact the resin. All solutions were removed via the bottom of the tube. "Stirred periodically" described in brief pulses N ₂ gas through the bottom frit; pulse duration from about 5 seconds and occur once every 30 seconds. The amino acid solution is usually used up to three weeks after preparation. The HATU solution was used within 5 days of preparation. DMF = dimethylformamide; HCTU = 2-(6-chloro-1H-benzotriazol-1-yl)-1,1,3,3-tetramethyl ;HATU=1-hexafluorophosphate 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide; NMM=N -methylmorpholine; Sieber=Fmoc-amino group- -3-yloxy, wherein "3-yloxy" describes the position and type of attachment to the polystyrene resin. The resin used was Merrifield polymer (polystyrene) with a Sieber linker (Fmoc protected at nitrogen); 100-200 mesh, 1% DVB, 0.71 mmol/g loading. The common amino acids used are listed below, wherein the side chain protecting groups are indicated in parentheses.

Fmoc-Ala-OH; Fmoc-Arg(Pbf)-OH; Fmoc-Asn(Trt)-OH; Fmoc-Asp(OtBu)-OH; Fmoc-Bzt-OH; Fmoc-Cys(Trt)-OH; Fmoc- Dab(Boc)-OH; Fmoc-Dap(Boc)-OH; Fmoc-Gln(Trt)-OH; Fmoc-Gly-OH; Fmoc-His(Trt)-OH; Fmoc-Ile-OH; Fmoc-Leu- OH; Fmoc-Lys(Boc)-OH; Fmoc-Nle-OH; Fmoc-[N-Me]Ala-OH; Fmoc-[N-Me]Nle-OH; Fmoc-Phe-OH; Fmoc-Pro-OH Fmoc-(D)-cis-Pro(4-OtBu)-OH; Fmoc-(D)-trans-Pro(4-OtBu)-OH; Fmoc-Sar-OH; Fmoc-Ser(tBu)-OH; Fmoc-Thr(tBu)-OH; Fmoc-Trp(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Val-OH.

The procedure of "Prelude Method A" describes an experiment conducted on a scale of 0.100 mmol, where the scale is determined by the amount of Sieber linker bound to the resin. This scale corresponds to about 140 mg of the above Sieber-Merrifield resin. All procedures can be scaled up by more than 0.100 mmol by adjusting the volume by a multiple of this size. Prior to amino acid coupling, all peptide synthesis sequences begin with a resin swelling procedure, described below as "resin swelling procedure." The N-terminal coupling of an amino acid with a primary amine uses the following "single coupling procedure". The N-terminal coupling of the amino acid with the secondary amine uses the following "secondary amine coupling procedure". Chloroacetyl group and peptide N-terminal The coupling is described by the "chloroethyl chloride coupling procedure" or "chloroacetic acid coupling procedure" detailed below.

Resin swelling procedure:

Merrifield Sieber resin (140 mg, 0.100 mmol) was added to a 40 mL polypropylene solid phase reaction vessel. The resin was washed (swelled) three times as follows: DMF (5.0 mL) and DCM (5.0 mL) were added to the reaction vessel, followed by bubbling the mixture from the bottom of the reaction vessel through N ₂ to periodically stir the mixture for 10 minutes, then passing the solvent through the frit. discharge.

Single coupling procedure:

Piperidine: DMF (20: 80 v/v, 5.0 mL) was added to a reaction vessel containing the resin from the previous step. The mixture was stirred periodically for 3 or 5 minutes and then the solution was discharged through the frit. Piperidine: DMF (20: 80 v/v, 5.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 or 5 minutes and then the solution was discharged through the frit. The resin was washed five times in succession as follows: DMF (4.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 60 seconds, after which the solution was discharged through the frit. Amino acid (0.2 M in DMF, 5.0 mL, 10 eq) was added to the reaction vessel, followed by addition of HATU or HCTU (0.2 M in DMF, 5.0 mL, 10 eq), and finally NMM (0.8M in DMF, 2.5 mL, 20 eq). The mixture was stirred periodically for 60 minutes, and then the reaction solution was discharged through a glass frit. The resin was washed four times continuously as follows: DFI (4.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 30 seconds, after which the solution was discharged through the frit. Acetic anhydride was added to the reaction vessel: DIEA:DMF solution (10:1:89 v/v/v, 5.0 mL). The mixture was stirred periodically for 10 minutes and then the solution was discharged through a frit. The resin was washed four times continuously as follows: DFI (4.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 90 seconds, after which the solution was discharged through the frit. The resulting resin was used directly in the next step.

Secondary amine coupling procedure:

Add piperidine:DMF to the reaction vessel containing the resin from the previous step (20:80 v/v, 5.0 mL). The mixture was stirred periodically for 3 or 5 minutes and then the solution was discharged through the frit. Piperidine: DMF (20: 80 v/v, 5.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 or 5 minutes and then the solution was discharged through the frit. The resin was washed five times continuously as follows: DFI (4.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 30 seconds, after which the solution was discharged through the frit. Amino acid (0.2 M in DMF, 2.5 mL, 5 eq) was added to the reaction vessel, followed by HATU (0.2M in DMF, 2.5 mL, 5 eq), and finally NMM (0.8 M in DMF, 1.5 mL) , 12eq). The mixture was stirred periodically for 300 minutes, and then the reaction solution was discharged through a glass frit. The resin was washed twice as follows: at each wash, DMF (4.0 mL) was added via the top of the vessel and the resulting mixture was stirred periodically for 30 seconds, then the solution was drained through the frit. Acetic anhydride was added to the reaction vessel: DIEA:DMF solution (10:1:89 v/v/v, 5.0 mL). The mixture was stirred periodically for 10 minutes and then the solution was discharged through a frit. The resin was washed four times continuously as follows: DFI (4.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 90 seconds, after which the solution was discharged through the frit. The resulting resin was used directly in the next step.

Custom amino acid coupling procedure:

Piperidine: DMF (20: 80 v/v, 5.0 mL) was added to a reaction vessel containing the resin from the previous step. The mixture was stirred periodically for 3 or 5 minutes and then the solution was discharged through the frit. Piperidine: DMF (20: 80 v/v, 5.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 or 5 minutes and then the solution was discharged through the frit. The resin was washed five times continuously as follows: DFI (4.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 30 seconds, after which the solution was discharged through the frit. Amino acid (0.2 M in DMF, 0.5 to 2.5 mL, 1 to 5 eq) was added to the reaction vessel, followed by addition of HATU (0.2 M in DMF, 0.5 to 2.5 mL, 1 to 5 eq), and finally DIPEA ( 0.8 M in DMF, 0.5 to 1.5 mL, 4 to 12 eq). The mixture is periodically stirred for 60 minutes to 600 minutes, and then the reaction solution is discharged through the glass frit. The resin was washed twice as follows: each wash At the time, DMF (2.0 mL) was added via the top of the vessel and the mixture was stirred periodically for 30 seconds, then the solution was drained through the glass frit. Acetic anhydride was added to the reaction vessel: DIEA:DMF solution (10:1:89 v/v/v, 5.0 mL). The mixture was stirred periodically for 10 minutes and then the solution was discharged through a frit. The resin was washed four times continuously as follows: DFI (4.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 90 seconds, after which the solution was discharged through the frit. The resulting resin was used directly in the next step.

Chloroacetyl chloride coupling procedure A:

Piperidine: DMF (20: 80 v/v, 5.0 mL) was added to a reaction vessel containing the resin from the previous step. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. Piperidine: DMF (20: 80 v/v, 5.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. The resin was washed five times continuously as follows: DFI (4.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 30 seconds, after which the solution was discharged through the frit. A solution of DIPEA (4.0 mmol, 0.699 mL, 40 eq) and chloroacetic acid chloride (2.0 mmol, 0.160 mL, 20 eq) in DMF was added. The mixture is periodically stirred for 12 to 18 hours, and then the solution is discharged through the glass frit. The resin was washed three times in succession as follows: DFI (4.0 mL) was added to the vessel at the top of each wash and the resulting mixture was periodically stirred for 90 seconds, after which the solution was discharged through the frit. The resin was washed four times in the following manner: On each wash, CH ₂ Cl ₂ (2.0 mL) was added to the vessel at the top and the resulting mixture was stirred periodically for 90 seconds, after which the solution was discharged through the glass frit.

Chloroacetic acid coupling procedure A:

Piperidine: DMF (20: 80 v/v, 5.0 mL) was added to a reaction vessel containing the resin from the previous step. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. Piperidine: DMF (20: 80 v/v, 5.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. The resin was washed five times continuously as follows: DFI (4.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 30 seconds, after which the solution was discharged through the frit. DMF (2.0 mL), chloroacetic acid (1.2 mmol, 113 mg, 12 eq), and N,N'-diisopropylcarbodiimide (1.2 mmol, 0.187 mL, 12 eq) were added to the reaction vessel. The mixture is periodically stirred for 12 to 18 hours, and then the solution is discharged through the glass frit. The resin was washed three times in succession as follows: DFI (4.0 mL) was added to the vessel at the top of each wash and the resulting mixture was periodically stirred for 90 seconds, after which the solution was discharged through the frit. The resin was washed four times in the following manner: On each wash, CH ₂ Cl ₂ (2.0 mL) was added to the vessel at the top and the resulting mixture was stirred periodically for 90 seconds, after which the solution was discharged through the glass frit.

CEM Method A:

All operations were performed on a CEM Liberty Microwave Peptide Synthesizer (CEM Corporation) according to automated operation. Unless otherwise noted, all procedures were performed in a 30 mL or 125 mL polypropylene tube with bottom frit in a CEM Discovery microwave unit. The tube is connected to the CEM Liberty synthesizer via the bottom and top of the tube. DMF and DCM can be added via the top and bottom of the tube, which are equally flushed along the sides of the tube. All solutions were removed through the bottom of the tube, but the resin was transferred from the top. "Periodic bubbling" describes a short bubbling of N ₂ gas through the bottom frit. The amino acid solution is usually used up to three weeks after preparation. The HATU solution was used within 5 days of preparation. DMF = dimethylformamide; HCTU = 2-(6-chloro-1H-benzotriazol-1-yl)-1,1,3,3-tetramethyl ;HATU=1-hexafluorophosphate 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide; DIPEA=II Isopropylethylamine; Sieber=Fmoc-Amino- -3-yloxy, wherein "3-yloxy" describes the position and type of attachment to the polystyrene resin. The resin used was Merrifield polymer (polystyrene) with a Sieber linker (Fmoc protected at nitrogen); 100-200 mesh, 1% DVB, 0.71 mmol/g loading. The common amino acids used are listed below, wherein the side chain protecting groups are indicated in parentheses.

Fmoc-Ala-OH; Fmoc-Arg(Pbf)-OH; Fmoc-Asn(Trt)-OH; Fmoc-Asp(OtBu)-OH; Fmoc-Bzt-OH; Fmoc-Cys(Trt)-OH; Fmoc- Dab(Boc)- OH; Fmoc-Dap(Boc)-OH; Fmoc-Gln(Trt)-OH; Fmoc-Gly-OH; Fmoc-His(Trt)-OH; Fmoc-Hyp(tBu)-OH; Fmoc-Ile-OH; Fmoc-Leu-OH; Fmoc-Lys(Boc)-OH; Fmoc-Nle-OH; Fmoc-Met-OH; Fmoc-[N-Me]Ala-OH; Fmoc-[N-Me]Nle-OH; Fmoc -Phe-OH; Fmoc-Sar-OH; Fmoc-Ser(tBu)-OH; Fmoc-Thr(tBu)-OH; Fmoc-Trp(Boc)-OH; Fmoc-Tyr(tBu) -OH; Fmoc-Val-OH.

The procedure for "CEM Method A" describes an experiment conducted on a scale of 0.100 mmol, where the scale is determined by the amount of Sieber linker bound to the resin. This scale corresponds to about 140 mg of the above Sieber-Merrifield resin. All procedures can be scaled up by more than 0.100 mmol by adjusting the volume by a multiple of this size. Prior to amino acid coupling, all peptide synthesis sequences begin with a resin swelling procedure, described below as "resin swelling procedure." The N-terminal coupling of an amino acid with a primary amine uses the following "single coupling procedure". The N-terminal coupling of the amino acid with the secondary amine uses the following "secondary amine coupling procedure". The coupling of the chloroethyl group with the N-terminus of the peptide is described by the "chloroethonium chloride coupling procedure" or the "chloroacetic acid coupling procedure" detailed above.

Resin swelling procedure:

Merrifield Sieber resin (140 mg, 0.100 mmol) was added to a 50 mL polypropylene conical tube. DMF (7 mL) was then added to the tube followed by DCM (7 mL). The resin is then transferred from the top of the vessel to the reaction vessel. This procedure is repeated twice more. DMF (7 mL) was added followed by DCM (7 mL). The resin was swollen by blowing N ₂ from the bottom of the reaction vessel for 15 minutes, and then the solvent was discharged through the glass frit.

Standard coupling procedure:

A piperidine:DMF solution (20:80 v/v, 5.0 mL) was added to the reaction vessel containing the resin from the previous step. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. A piperidine:DMF solution (20:80 v/v, 5.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. The resin was washed three times as follows: DMF (7 mL) was washed from the top, then washed from bottom with DMF (7 mL) and finally washed from top with DMF (7 mL). Amino acid (0.2 M in DMF, 2.5 mL, 5 eq), HATU (0.5 M in DMF, 1.0 mL, 5 eq) and DIPEA (2M in NMP, 0.5 mL, 10 eq) was added. A mixture of all amino acids was bubbled through N ₂ at 75 ° C for 5 minutes (but where Fmoc-Cys(Trt)-OH and Fmoc-His(Trt)-OH were coupled at 50 ° C), and the reaction solution was passed through the frit. discharge. The resin was washed three times as follows: DMF (7 mL) was washed from the top, then washed from bottom with DMF (7 mL) and finally washed from top with DMF (7 mL). Acetic anhydride was added to the reaction vessel: DIEA:DMF solution (10:1:89 v/v/v, 5.0 mL). The mixture was periodically bubbled at 65 ° C for 2 minutes, and then the solution was discharged through a frit. The resin was washed three times as follows: DMF (7 mL) was washed from the top, then washed from bottom with DMF (7 mL) and finally washed from top with DMF (7 mL). The resulting resin was used directly in the next step.

Double pair coupling procedure

A piperidine:DMF solution (20:80 v/v, 5.0 mL) was added to the reaction vessel containing the resin from the previous step. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. A piperidine:DMF solution (20:80 v/v, 5.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. The resin was washed three times as follows: DMF (7 mL) was washed from the top, then washed from bottom with DMF (7 mL) and finally washed from top with DMF (7 mL). Amino acid (0.2 M in DMF, 2.5 mL, 5 eq), HATU (0.5 M in DMF, 1.0 mL, 5 eq) and DIPEA (2M in NMP, 0.5 mL, 10 eq) was added. A mixture of all amino acids was bubbled through N ₂ at 75 ° C for 5 minutes (but where Fmoc-Cys(Trt)-OH and Fmoc-His(Trt)-OH were coupled at 50 ° C), and the reaction solution was passed through the frit. discharge. The resin was washed three times as follows: DMF (7 mL) was washed from the top, then washed from bottom with DMF (7 mL) and finally washed from top with DMF (7 mL). Amino acid (0.2 M in DMF, 2.5 mL, 5 eq), HATU (0.5 M in DMF, 1.0 mL, 5 eq) and DIPEA (2M in NMP, 0.5 mL, 10 eq) was added. A mixture of all amino acids was bubbled through N ₂ at 75 ° C for 5 minutes (but where Fmoc-Cys(Trt)-OH and Fmoc-His(Trt)-OH were coupled at 50 ° C), and the reaction solution was passed through the frit. discharge. The resin was washed three times as follows: DMF (7 mL) was washed from the top, then washed from bottom with DMF (7 mL) and finally washed from top with DMF (7 mL). Acetic anhydride was added to the reaction vessel: DIEA:DMF solution (10:1:89 v/v/v, 5.0 mL). The mixture was periodically bubbled at 65 ° C for 2 minutes, and then the solution was discharged through a frit. The resin was washed three times as follows: DMF (7 mL) was washed from the top, then washed from bottom with DMF (7 mL) and finally washed from top with DMF (7 mL). The resulting resin was used directly in the next step.

Custom amino acid coupling procedure:

A piperidine:DMF solution (20:80 v/v, 5.0 mL) was added to the reaction vessel containing the resin from the previous step. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. A piperidine:DMF solution (20:80 v/v, 5.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. The resin was washed three times as follows: DMF (7 mL) was washed from the top, then washed from bottom with DMF (7 mL) and finally washed from top with DMF (7 mL). A solution of HATU (2.5 eq to 10 eq) in amino acid (1.25 mL to 5 mL, 2.5 eq to 10 eq) was added to the reaction vessel, and finally DIPEA (2M in NMP, 0.5 mL to 1 mL, 20 eq) was added. The mixture was mixed by bubbling N ₂ at 25 ° C to 75 ° C for 5 minutes to 2 hours, and then the reaction solution was discharged through a glass frit. The resin was washed three times as follows: DMF (7 mL) was washed from the top, then washed from bottom with DMF (7 mL) and finally washed from top with DMF (7 mL). Acetic anhydride was added to the reaction vessel: DIEA:DMF solution (10:1:89 v/v/v, 5.0 mL). The mixture was periodically bubbled at 65 ° C for 2 minutes, and then the solution was discharged through a frit. The resin was washed three times as follows: DMF (7 mL) was washed from the top, then washed from bottom with DMF (7 mL) and finally washed from top with DMF (7 mL). The resulting resin was used directly in the next step.

Symphony Method A:

All operations were performed on a Symphony peptide synthesizer (Protein Technologies) according to automated operation. Unless otherwise noted, all procedures were performed in a Symphony polypropylene tube with a bottom frit. The tube is connected to the Symphony peptide synthesizer via the bottom and top of the tube. All solvents DMF, DCM, amino acids and reagents were added via the bottom of the tube and passed up through the frit to contact the resin. All solutions were removed via the bottom of the tube. "Stirred periodically" described in brief pulses N ₂ gas through the bottom frit; pulse duration from about 5 seconds and occur once every 15 seconds. The amino acid solution is usually used up to three weeks after preparation. The HATU solution was used within 5 days of preparation. DMF = dimethylformamide; HCTU = 2-(6-chloro-1H-benzotriazol-1-yl)-1,1,3,3-tetramethyl ;HATU=1-hexafluorophosphate 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide; NMM=N -methylmorpholine; DIPEA = diisopropylethylamine; Sieber = Fmoc-amino group - -3-yloxy, wherein "3-yloxy" describes the position and type of attachment to the polystyrene resin. The resin used was Merrifield polymer (polystyrene) with a Sieber linker (Fmoc protected at nitrogen); 100-200 mesh, 1% DVB, 0.71 mmol/g loading. Other common acid-sensitive resins such as Rink or functionalized chlorotrityl resins can also be used in the synthesis. The common amino acids used are listed below, wherein the side chain protecting groups are indicated in parentheses.

Fmoc-Ala-OH; Fmoc-Arg(Pbf)-OH; Fmoc-Asn(Trt)-OH; Fmoc-Asp(OtBu)-OH; Fmoc-Bzt-OH; Fmoc-Cys(Trt)-OH; Fmoc- Dab(Boc)-OH; Fmoc-Dap(Boc)-OH; Fmoc-Gln(Trt)-OH; Fmoc-Gly-OH; Fmoc-His(Trt)-OH; Fmoc-Hyp(tBu)-OH; Fmoc -Ile-OH; Fmoc-Leu-OH; Fmoc-Lys(Boc)-OH; Fmoc-Nle-OH; Fmoc-Met-OH; Fmoc-[N-Me]Ala-OH; Fmoc-[N-Me] Nle-OH; Fmoc-Phe-OH; Fmoc-Pro-OH; Fmoc-Sar-OH; Fmoc-Ser(tBu)-OH; Fmoc-Thr(tBu)-OH; Fmoc-Trp(Boc)-OH; Fmoc -Tyr(tBu)-OH; Fmoc-Val-OH.

The procedure for "Symphony Method A" describes an experiment conducted on a scale of 0.050-0.100 mmol, where the scale is determined by the amount of Sieber linker bound to the resin. This regulation The mold corresponds to about 70-140 mg of the above Sieber-Merrifield resin. All procedures can be scaled up by more than 0.050-0.100 mmol by adjusting the volume by a multiple of this size. Prior to amino acid coupling, all peptide synthesis sequences begin with a resin swelling procedure, which is described below as a "swelling procedure." The N-terminal coupling of the amino acid with the primary amine uses the following "standard coupling procedure". The N-terminal coupling of the amino acid with the secondary amine uses "double coupling", and the conventional amino acid is coupled by manual addition of an amino acid ("blank coupling" below).

Swelling procedure:

Merrifield Sieber resin (70 mg, 0.050 mmol or 140 mg, 0.100 mmol) was added to a Symphony polypropylene solid phase reaction vessel. The resin was washed (swelled) three times as follows: DMF (2.5 mL) was added to the reaction vessel, followed by bubbling the mixture from the bottom of the reaction vessel through N ₂ to periodically stir the mixture for 10 minutes, and then the solvent was discharged through the glass frit.

Standard coupling procedure:

The resin was washed three times as follows: DMF (2.5 mL) was added to the reaction vessel, followed by bubbling the mixture from the bottom of the reaction vessel via N ₂ to periodically stir the mixture for 30 seconds, and then the solvent was discharged through the glass frit. Piperidine: DMF (20: 80 v/v, 2.5 mL) was added to the reaction vessel. The mixture was stirred periodically for 5 minutes and then the solution was discharged through the frit. This procedure is repeated again. The resin was washed 6 times as follows: DMF (2.5 mL) was added via the bottom of the vessel with each wash and the resulting mixture was periodically stirred for 30 seconds, after which the solution was discharged through the frit. Amino acid (0.2 M in DMF, 1.25 mL, 5 eq) was added to the reaction vessel, followed by HATU (0.2M in DMF, 1.25 mL, 5 eq), and finally NMM (0.8M in DMF, 1.25 mL) , 10eq). The mixture was stirred periodically for 10 minutes, and then the reaction solution was discharged through a glass frit. The resin was washed with DMF (6.25 mL) added via the bottom of the vessel and the mixture was stirred periodically for 30 seconds, then the solution was drained through the frit. Amino acid (0.2 M in DMF, 1.25 mL, 5 eq) was added to the reaction vessel, followed by HATU (0.2M in DMF, 1.25 mL, 5 eq), and finally NMM (0.8M in DMF, 1.25 mL) , 10eq). The mixture was stirred periodically for 10 minutes, and then the reaction solution was discharged through a glass frit. The resin was washed three times in succession: DMF (2.5 mL) was added via the bottom of the vessel with each wash and the resulting mixture was periodically stirred for 30 seconds, after which the solution was discharged through the frit. The resulting resin was used directly in the next step.

Secondary amine coupling procedure:

The resin was washed three times as follows: DMF (2.5 mL) was added to the reaction vessel, followed by bubbling the mixture from the bottom of the reaction vessel via N ₂ to periodically stir the mixture for 30 seconds, and then the solvent was discharged through the glass frit. Piperidine: DMF (20: 80 v/v, 2.5 mL) was added to the reaction vessel. The mixture was stirred periodically for 5 minutes and then the solution was discharged through the frit. This procedure is repeated again. The resin was washed 6 times as follows: DMF (2.5 mL) was added via the bottom of the vessel with each wash and the resulting mixture was periodically stirred for 30 seconds, after which the solution was discharged through the frit. Amino acid (0.2 M in DMF, 1.25 mL, 5 eq) was added to the reaction vessel, followed by HATU (0.2M in DMF, 1.25 mL, 5 eq), and finally NMM (0.8M in DMF, 1.25 mL) , 10eq). The mixture was stirred periodically for 300 minutes, and then the reaction solution was discharged through a glass frit. The resin was washed with DMF (6.25 mL) added via the bottom of the vessel and the mixture was stirred periodically for 30 seconds, then the solution was drained through the frit. Amino acid (0.2 M in DMF, 1.25 mL, 5 eq) was added to the reaction vessel, followed by HATU (0.2M in DMF, 1.25 mL, 5 eq), and finally NMM (0.8M in DMF, 1.25 mL) , 10eq). The mixture was stirred periodically for 300 minutes, and then the reaction solution was discharged through a glass frit. The resin was washed three times in succession: DMF (2.5 mL) was added via the bottom of the vessel with each wash and the resulting mixture was periodically stirred for 30 seconds, after which the solution was discharged through the frit. The resulting resin was used directly in the next step.

Symphony Method B Final capping procedure:

The resin was washed three times as follows: DMF (2.5 mL) was added to the reaction vessel, followed by bubbling the mixture from the bottom of the reaction vessel via N ₂ to periodically stir the mixture for 30 seconds, and then the solvent was discharged through the glass frit. Piperidine: DMF (20: 80 v/v, 2.5 mL) was added to the reaction vessel. The mixture was stirred periodically for 2.5 minutes and then the solution was discharged through a frit. The resin was washed 6 times as follows: DMF (2.5 mL) was added via the bottom of the vessel with each wash and the resulting mixture was periodically stirred for 30 seconds, after which the solution was discharged through the frit. NMM (0.8 M in DMF, 1.25 mL, 10 eq) was then added to the reaction mixture, then chloroacetic acid (0.4M in DMF, 1.25 mL, 10 eq). The mixture was stirred periodically for 15 minutes, and then the reaction solution was discharged through a frit. The resin was washed with DMF (6.25 mL) added via the bottom of the vessel and the mixture was stirred periodically for 30 seconds, then the solution was drained through the frit. NMM (0.8 M in DMF, 1.25 mL, 10 eq) was then added to the reaction mixture, then chloroacetic acid (0.4M in DMF, 1.25 mL, 10 eq). The mixture was stirred periodically for 15 minutes, and then the reaction solution was discharged through a frit. The resin was washed 6 times as follows: DMF (2.5 mL) was added via the bottom of the vessel and the mixture was stirred periodically for 30 seconds, then the solution was drained through the frit. Ac ₂ O/DIPEA/DMF (v/v/v 1:1:3, 2.5 mL) was added to the reaction vessel, and the mixture was stirred periodically for 10 minutes, and then the reaction solution was discharged through a glass frit. The resin was washed six times in succession as follows: DMF (2.5 mL) was added via the bottom of the vessel at each wash and the resulting mixture was periodically stirred for 30 seconds, after which the solution was discharged through the frit. The resin was washed four times as follows: DCM (2.5 mL) was added via the bottom of the vessel with each wash and the resulting mixture was periodically stirred for 30 seconds, after which the solution was discharged through the frit. The resulting resin was then dried with a stream of nitrogen for 10 minutes.

N-methylation on-resin (Turner, RA; Hauksson, NE; Gipe, JH; Lokey, RS Org. Lett. 2013, 15 (19), 5012-5015): Unless otherwise noted, all operations were performed manually. The procedure for "N-methylation on resin" describes an experiment conducted on a scale of 0.100 mmol, wherein the scale is determined by the amount of Sieber linker bound to the resin used to produce the peptide. This scale is not directly determined by the amount of peptide used in the program. The procedure can be expanded by more than 0.100 mmol by adjusting the volume in multiples of this size.

The resin was transferred to a 25 mL syringe equipped with frit. Adding pipe to the resin Pyridine: DMF (20:80 v/v, 5.0 mL). The mixture was shaken for 3 minutes and then the solution was discharged through a frit. The resin was washed 3 times with DMF (4.0 mL). Piperidine: DMF (20: 80 v/v, 4.0 mL) was added to the reaction vessel. The mixture was shaken for 3 minutes and then the solution was discharged through a frit. The resin was washed six times in the following manner: DMF (4.0 mL) was added three times and the resulting mixture was shaken for 3 seconds, then the solution was drained through the glass frit, then DCM (4.0 mL) was added three times and the mixture was shaken for 3 s. The glass frit is discharged.

The resin was suspended in DMF (2.0 mL) and ethyl trifluoroacetate (0.119 ml, 1.00 mmol), and 1,8-diazabicyclo[5.4.0]undec-7-ene (0.181 ml, 1.20 mmol) . The mixture was placed on the shaker for 60 minutes. The solution is allowed to drain through the frit. The resin was washed six times in the following manner: DMF (4.0 mL) was added three times and the resulting mixture was shaken for 3 seconds, then the solution was drained through the glass frit, then DCM (4.0 mL) was added three times and the mixture was shaken for 3 s. The glass frit is discharged.

The resin was washed 3 times with anhydrous THF (2.0 mL) to remove any residual water. To a dried 4.0 mL vial was added THF (1.0 mL), triphenylphosphine (131 mg, 0.500 mmol) / dried 4 Å molecular sieve (20 mg). The turbid solution was transferred to the resin and diisopropyl azodicarboxylate (0.097 mL, 0.5 mmol) was slowly added. The resin was shaken for 15 minutes. The solution was drained through a frit and the resin was washed 3 times with dry THF (2.0 mL) to remove any residual water. To a dried 4.0 mL vial was added THF (1.0 mL), triphenylphosphine (131 mg, 0.500 mmol) / dried 4 Å molecular sieve (20 mg). The turbid solution was transferred to the resin and diisopropyl azodicarboxylate (0.097 mL, 0.5 mmol) was slowly added. The resin was shaken for 15 minutes. The solution is allowed to drain through the frit. The resin was washed six times in the following manner: DMF (4.0 mL) was added three times and the resulting mixture was shaken for 3 seconds, then the solution was drained through the glass frit, then DCM (4.0 mL) was added three times and the mixture was shaken for 3 s. The glass frit is discharged.

The resin was suspended in ethanol (1.0 mL) and THF (1.0 mL), and sodium borohydride (37.8 mg, 1.00 mmol) was added. The mixture was mixed on a shaker for 30 minutes. The solution was drained through the frit and the resin was washed six times as follows: DMF (4.0 mL) was added 3 times and the resulting The mixture was shaken for 3 seconds, then the solution was drained through a frit, then DCM (4.0 mL) was added three times and the mixture was shaken for 3 s, then the solution was drained through glass frit.

Overall removal of the protective method B:

All operations are performed manually unless otherwise noted. The procedure for "Total Removal of Protecting Group Method B" describes an experiment conducted on a scale of 0.100 mmol, where the scale is determined by the amount of Sieber linker bound to the resin. The procedure can be expanded by more than 0.100 mmol by adjusting the volume in multiples of this size. A "removal protecting group solution" was prepared using trifluoroacetic acid: triisopropyl decane: dithiothreitol (94:3:3 v:v:w). The resin was removed from the reaction vessel and transferred to a 25 mL syringe equipped with frit. A "removal protecting group solution" (5.0 mL) was added to the syringe. The mixture was mixed in a shaker for 5 minutes. The solution was filtered and diluted in diethyl ether (30 mL). The precipitated solid was centrifuged for 3 minutes. The supernatant solution was decanted and the solid was resuspended in diethyl ether (25 mL). The suspension was centrifuged for 3 minutes. The supernatant was decanted and the remaining solid was suspended in diethyl ether (25 mL). The suspension was centrifuged for 3 minutes. The supernatant was decanted and the remaining solid was dried under high vacuum. The crude peptide was obtained as a white to off-white solid.

Overall removal of protecting group method G

All operations are performed manually unless otherwise noted. The procedure for "Total Removal of Protecting Group Method G" describes an experiment conducted on a scale of 0.50 or 0.100 mmol, where the scale is determined by the amount of Sieber linker bound to the resin. The procedure can be expanded by more than 0.100 mmol by adjusting the volume in multiples of this size. A "removal protecting group solution" was prepared using trifluoroacetic acid: triisopropyl decane: water (95:2.5:2.5 v:v:v). A "removal protecting group solution" (2.5 mL) was added to the resin. The mixture was stirred for 5 minutes. The solution was filtered and the filtrate was taken in cold diethyl ether (40 mL). The resin was treated with an additional 2.5 mL of "Removal of the protecting group solution" for 2 minutes and the filtrate was added to cold diethyl ether from the previous treatment. The precipitated solid was collected by EtOAc (EtOAc) elute

Cyclization method C:

All operations are performed manually unless otherwise noted. The procedure of "Cycyclization Method C" describes an experiment conducted on a scale of 0.100 mmol, wherein the scale is determined by the amount of Sieber linker bound to the resin used to produce the peptide. This scale is not directly determined by the amount of peptide used in the program. The procedure can be expanded by more than 0.100 mmol by adjusting the volume in multiples of this size. The crude peptide solid was dissolved in a solution of acetonitrile: 0.1 M aqueous ammonium bicarbonate (11 mL: 24 mL), and then the solution was carefully adjusted to pH = 8.5-9.0 using aqueous NaOH (1.0M). The solution is then mixed using an oscillator for 12 to 18 hours. The reaction solution was concentrated and the residue was dissolved in acetonitrile: water. This solution was subjected to reverse phase HPLC purification to give the desired cyclic peptide.

Preparation of racemic 2-(((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(1-(t-butoxycarbonyl)-1H-pyrrolo[2,3- b]pyridin-3-yl)propionic acid

(Robison, MM and Robison, BL J. Am.. Chem. Soc. , 1955, 77 , 457-459)

step 1:

3-(dimethylaminomethyl)-7-azagramine (3.5 g, 19.97 mmol), diethyl acetaminomalonate (4.34 g, 19.97 mmol) and xylene (35ml) mixture Sodium hydroxide powder (0.080 g, 1.997 mmol) was treated and stirred under reflux for 15 h under nitrogen. The hot solution was filtered to give a yellow filtrate. Upon cooling to room temperature, a yellow solid precipitated in the filtrate. The solid block was suspended in benzene (40 mL) and filtered. The collected solid was washed with hexane (2×100 mL) to afford 2-((1H-pyrrolo[2,3-b]pyridin-3-yl)methyl)-2-acetamide as a white solid. Diethyl malonate (3.5 g, 50.4%).

Analytical LCMS Condition A: Retention time = 0.79 min; ESI-MS (+) m/z 348.3 (M+H).

Step 2:

Diethyl 2-((1H-pyrrolo[2,3-b]pyridin-3-yl)methyl)-2-acetamidomalonate (3.5 g, 10.08 mmol) and 37% hydrochloric acid (30 mL) The mixture was refluxed for 15 hours and concentrated to 10 mL. The product was treated with EtOAc (5 mL) and EtOAc evaporated This was redissolved in 15% NH ₄ OH 7 and the solution was then diluted with water until the pH (20mL). The precipitated white solid was collected by filtration, washed with water andEtOAc and dried to afford 2-amino 3-(1H-pyrrolo[2,3-b]pyridin-3-yl)propanoic acid as a white solid ( 1.5g, 72.5%).

Analytical LCMS Condition A: Retention time = 0.29 min; ESI-MS (+) m/z 206.0 (M+H).

Step 3:

2-Amino-3-(1H-pyrrolo[2,3-b]pyridin-3-yl)propanoic acid (1.3 g, 6.33 mmol) and triethylamine (1.766 ml, 12.67 mmol) in acetonitrile (20 ml) The solution in water (15 ml) was treated with EtOAc (EtOAc: EtOAc (EtOAc) The mixture was concentrated to dryness to give a white solid. The solid was treated with 1M EtOAc (EtOAc) (EtOAc)EtOAc. The obtained product was suspended in anhydrous MeOH (50 mL) and 4M HCl / dioxane (10 mL) and the solution was refluxed for one hour. The mixture was concentrated under reduced pressure and the residue was partitioned between EtOAc and filtered with saturated aqueous NaHCO _3. The EtOAc was washed with EtOAc (EtOAc) (HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH Methyl 3-(1H-pyrrolo[2,3-b]pyridin-3-yl)propanoate (1.0 g, 28% yield).

Analytical LCMS Condition A: Retention time = 0.81 min; ESI-MS (+) m/z 442.5 (M+H).

Step 4:

2-((((9H-茀-9-yl)methoxy)carbonyl)amino)-3-(1H-pyrrolo[2,3-b]pyridin-3-yl)propanoic acid methyl ester (1.0 A solution of g, 2.265 mmol, EtOAc (EtOAc) (EtOAc) A solution of Boc ₂ O (0.789 mL, 3.40 mmol) in THF (5 mL). The reaction mixture was diluted with EtOAc (150mL), washed with saturated NH ₄ Cl (3 × 50mL) and brine (50mL), and concentrated under reduced pressure over sodium sulfate. The crude product was purified by flash chromatography using EtOAc (EtOAc) elute Methoxy)carbonyl)amino)-3-methoxy-3-oxopropyl)-1H-pyrrolo[2,3-b]pyridine-1-carboxylic acid tert-butyl ester (0.85 g, 69 %Yield).

Analytical LCMS Condition A: Retention time = 1.01 min; ESI-MS (+) m/z 542.5 (M+H).

Step 5:

3-(2-(((9H-茀-9-yl)methoxy)carbonyl)amino)-3-methoxy-3-oxopropyl)-1H-pyrrolo[2,3 -b] a solution of pyridine-1-carboxylic acid tert-butyl ester (200 mg, 0.369 mmol) in anhydrous 1,2-dichloroethane (5 mL) with trimethyltin hydroxide (200 mg, 1.108 mmol) and mixture Stir at 65 ° C for 1 hour. The mixture was concentrated with EtOAc (EtOAc m. -茀-9-yl)methoxy)carbonyl)amino)-3-(1-(t-butoxycarbonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)propanoic acid. This material was used without further purification. ^{1 H NMR (400MHz, DMSO-} d 6) δ 8.40-8.37 (m, 1H), 8.12-8.05 (m, 1H), 7.90-7.75 (m, 2H), 7.70-7.55 (m, 2H), 7.43- 7.35 (m, 2H), 7.30-7.18 (m, 4H), 4.37-4.29 (m, 1H), 4.16-4.12 (m, 3H), 3.23 - 3.15 (m, 1H), 3.07-2.98 (m, 1H) ), 1.60 (s, 9H).

Analytical LCMS Condition A: Retention time = 0.93 min; ESI-MS (+) m/z 528.4 (M+H).

Preparation of Example 3210

Sieber resin (140 mg, 0.100 mmol) was added to a 50 mL polypropylene tube, and the tube was placed on a CEM Liberty microwave peptide synthesizer. Then execute the following procedure in sequence: follow "CEM Method A: Resin Swelling Procedure"; use Fmoc-Gly-OH, follow "CEM Method A: Standard Coupling Procedure"; use Fmoc-Cys(Trt)-OH, follow "CEM Method A : Standard coupling procedure"; use Fmoc-Leu-OH, follow "CEM Method A: Standard Coupling Procedure"; use Fmoc-Tyr(tBu)-OH, follow "CEM Method A: Standard Coupling Procedure"; use Fmoc-Trp ( Boc)-OH, follow "CEM Method A: Standard Coupling Procedure"; Fmoc-D-Pro (5,5-di-Me) is artificially coupled as follows: Add Fmoc-D-Pro to the peptidyl resin (5, A 5 mL solution of 5-di-Me)-OH (1.2 eq), HATU (1.2 eq), and DIEA (2.5 eq). The resin suspension was stirred for 16 hours. The resin was washed with DMF (3.times.5 mL), DCM (3.times.5mL) and washed once again with DMF (5mL). Then proceed on the CEM synthesizer Continued synthesis.

Use Fmoc-Tyr(tBu)-OH, follow "CEM Method A: Standard Coupling Procedure"; use Fmoc-[N-Me]Phe-OH, follow "CEM Method A: Standard Coupling Procedure"; use 10eq Fmoc-Val- OH (10 minutes at 75 ° C, then 2 hours at room temperature), follow "CEM Method A: Custom Amino Acid Coupling Procedure "; use Fmoc-Asp(OtBu)-OH, follow "CEM Method A: Standard coupling procedure"; use Fmoc-Sar-OH, follow "CEM Method A: Standard Coupling Procedure"; use 5eq Fmoc-[N-Me]Nle-OH (for 10 minutes), follow "CEM Method A: Custom Amine Base acid coupling procedure ; using 5 eq of Fmoc-[N-Me]Phe-OH (for 10 minutes), following "CEM Method A: Custom Amino Acid Coupling Procedure "; using 5 eq of Fmoc-Phe-OH (for 10 minutes) ), follow the "CEM Method A: Custom Amino Acid Coupling Procedure "; use, follow "Prelude Method A: Chloroacetate Chloride Coupling Procedure A"; use, follow "Total Removal of Protective Group Method B"; use, follow "Circularization Method C".

The crude material was purified via preparative LC/MS using the following conditions: column: Phenomenex Luna 20x250 5μ particles; mobile phase A: water + 0.1% TFA; mobile phase B: acetonitrile + 0.1% TFA; gradient: 30-80% B It lasted for 50 minutes, then held at 80% B for 5 minutes; flow: 15 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 1.1 mg, and the purity was estimated to be 99% based on "analytical HPLC Condition B" using 35% to 85% Buffer B (30 minutes).

Analytical LCMS condition A: retention time = 1.55min; ESI-MS (+ ) m / z 925.8 (M + 2H). ESI-HRMS (+) m/z: Calculated: 925.4547 (M+2H).

Experimental value: 925.4551 (M+2H).

Preparation of Example 3211

Example 3211 was prepared following the general synthetic sequence described for the preparation of Example 3210, which consisted of the following general procedure: "CEM Method A: Resin Swelling Procedure", "CEM Method A: Standard Coupling Procedure", " Customization ""Amino acid coupling procedure ", "CEM method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 15-65% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.4 mg and the purity was estimated to be 97% based on LCMS analysis using "Analysis Conditions D and E".

Analytical LCMS conditions D: retention time = 1.60 min; ESI-MS (+) m/z s .

Analytical LCMS Condition E: Retention time = 1.82 min; ESI-MS (+) m/z .

Preparation of Example 3212

Sieber resin (140 mg, 0.100 mmol) was added to a 40 mL polypropylene solid phase reaction vessel, and the reaction vessel was placed on a Prelude peptide synthesizer. Then execute the following procedures in sequence: use, follow "Prelude Method A: Resin Swelling Procedure"; use Fmoc-Gly-OH, follow "Prelude Method A: Single Coupling Procedure"; use Fmoc-Cys(Trt)-OH, follow "Prelude Method A: Single coupling procedure"; using Fmoc-Leu-OH, following "Prelude Method A: Single Coupling Procedure"; using Fmoc-Phe(CH ₂ NH ₂ )-OH, following "Prelude Method A: Single Coupling Procedure"; Use Fmoc-Trp(Boc)-OH, follow "Prelude Method A: Single Coupling Procedure"; use Fmoc-Sar-OH, follow "Prelude Method A: Single Coupling Procedure"; use Fmoc-Asp(OtBu)-OH, follow "Prelude Method A: Single Coupling Procedure"; use Fmoc-[N-Me]Phe-OH, follow "Prelude Method A: Single Coupling Procedure"; use Fmoc-Val-OH, follow "Prelude Method A: Secondary Amine Coupling"Procedure; use Fmoc-Asn(Trt)-OH, follow "Prelude Method A: Single Coupling Procedure"; use Fmoc-Sar-OH, follow "Prelude Method A: Secondary Amine Coupling Procedure "; use Fmoc-[N- Me]Nle-OH, follow "Prelude Method A: Secondary Amine Coupling Procedure "; use Fmoc-[N- Me]Phe-OH, follow “Prelude Method A: Secondary Amine Coupling Procedure ”; use Fmoc-Phe-OH, follow “Prelude Method A: Secondary Amine Coupling Procedure ”; use, follow “Prelude Method A: Chloroacetamidine” Chlorine coupling procedure A"; use, follow the "total removal of protecting group method B"; use, follow the "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-60% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.8 mg, and the purity was estimated to be 97% based on LCMS analysis using "Analysis Conditions D and E".

Analytical LCMS conditions D: retention time = 1.62 min; ESI-MS (+) m/z 880.7 (M+2H).

Analytical LCMS Condition E: Retention time = 1.61 min; ESI-MS (+) m/z 880.7 (M+2H).

Preparation of Example 3213

Example 3213 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 15-65% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.5 mg and the purity was estimated to be 99% based on LCMS analysis using "Analysis Conditions D and E".

Analytical LCMS conditions D: retention time = 1.60 min; ESI-MS (+) m/z 887.9 (M+2H).

Analytical LCMS Condition E: Retention time = 1.60 min; ESI-MS (+) m/z 887.7 (M+2H).

Preparation of Example 3216

Example 3216 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 15-65% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 25-70% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 1.3 mg and the purity was estimated to be 96% based on LCMS analysis using "Analysis Conditions D and E".

Analytical LCMS conditions D: retention time = 1.69 min; ESI-MS (+) m/z 949.2 (M+2H).

Analytical LCMS Condition E: Retention time = 1.70 min; ESI-MS (+) m/z 949.2 (M+2H).

Preparation of Example 3217

Example 3217 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 35-85% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.2 mg and the purity was estimated to be 98% based on LCMS analysis using "Analysis Conditions D and E".

Analytical LCMS conditions D: retention time = 2.43 min; ESI-MS (+) m/z 956.8 (M+2H).

Analytical LCMS conditions E: residence time = 2.43 min; ESI-MS (+) m/z 957.0 (M+2H).

ESI-HRMS (+) m/z: Calculated: 956.4613 (M+2H).

Found: 956.4604 (M+2H).

Preparation of Example 3218

Example 3218 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 20-70% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.6 mg and the purity was estimated to be 94% based on LCMS analysis using "Analysis Conditions D and E".

Analytical LCMS conditions D: retention time = 2.12 min; ESI-MS (+) m/z 957.2 (M+2H).

Analytical LCMS Condition E: Retention time = 1.83 min; ESI-MS (+) m/z 957.4 (M+2H).

ESI-HRMS (+) m/z: Calculated: 956.9590 (M+2H).

Found: 95.9582 (M+2H).

Preparation of Example 3219

Example 3219 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 30-80% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.9 mg and the purity was estimated based on LCMS analysis using "Analysis Conditions D and E". 96%.

Analytical LCMS conditions D: retention time = 2.26 min; ESI-MS (+) m/z 957.9 (M+2H).

Analytical LCMS conditions E: retention time = 2.26 min; ESI-MS (+) m/z 958.0 (M+2H).

ESI-HRMS (+) m/z: Calculated: 95.445.10 (M+2H).

Found: 956.4493 (M+2H).

Preparation of Example 3220

Example 3220 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 25-75% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. Consolidation The fractions of the desired product were dried and dried by centrifugal evaporation. The yield of the product was 8.4 mg and the purity was estimated to be 92% based on LCMS analysis using "Analysis Conditions D and E".

Analytical LCMS conditions D: retention time = 1.71 min; ESI-MS (+) m/z 932.0 (M+2H).

Analytical LCMS conditions E: residence time = 1.94 min; ESI-MS (+) m/z 932.0 (M+2H).

ESI-HRMS (+) m/z: calc.

Found: 931.4536 (M+2H).

Preparation of Example 3221

Example 3221 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% Trifluoroacetic acid; mobile phase B: 95:5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 30-80% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.2 mg, and the purity was estimated to be 94% according to LCMS analysis using "Analysis Conditions D and E".

Analytical LCMS conditions D: residence time = 2.22 min; ESI-MS (+) m/z 942.1 (M+2H).

Analytical LCMS conditions E: residence time = 2.20 min; ESI-MS (+) m/z 943.2 (M+2H).

ESI-HRMS (+) m/z: Calculated: 942.4457 (M+2H).

Found: 942.4445 (M+2H).

Preparation of Example 3222

Example 3222 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: Phenomenex Luna 20x250 5u particles; mobile phase A: water + 0.1% TFA; mobile phase B: acetonitrile + 0.1% TFA; gradient: .35-95% B lasted 50 minutes, followed by 5 minutes at 95% B; flow: 15 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.9 mg and the purity was estimated to be 99% based on "Analytical HPLC Condition B" using 35% to 85% Buffer B (30 minutes).

Analytical LCMS Condition A: Retention time = 1.65 min; ESI-MS (+) m/z 966.1 (M+2H).

ESI-HRMS (+) m/z: Calculated: 965.4484 (M+2H).

Found: 965.4473 (M+2H).

Preparation of Example 3223

Example 3223 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: Phenomenex Luna 20x250 5u particles; mobile phase A: water + 0.1% TFA; mobile phase B: acetonitrile + 0.1% TFA; gradient: 35-95% B for 50 minutes, followed by 95% B for 5 minutes; flow: 15 mL / Min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.1 mg, and the purity was estimated to be 98% based on "analytical HPLC Condition B" using 35% to 85% Buffer B (30 minutes).

Analytical LCMS Condition A: Retention time = 1.65 min; ESI-MS (+) m/z 966.8 (M+2H).

ESI-HRMS (+) m/z: Calculated: 966.4381 (M+2H).

Found: 966.4375 (M+2H).

Preparation of Example 3224

Example 3224 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

Mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; Flow rate: 20 mL/min. Combine the fractions containing the desired product and steam by centrifugation Hair is dried. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 25-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.7 mg, and the purity was estimated to be 97% based on LCMS analysis using "analytical LCMS Condition E".

Analytical LCMS conditions E: residence time = 1.93 min; ESI-MS (+) m/z 939.31 (M+2H).

ESI-HRMS (+) m/z: calc.

Experimental value: 939.4322 (M+2H).

Preparation of Example 3225

Example 3225 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.2 mg, and the purity was estimated to be 96% based on LCMS analysis using "Analysis Conditions E and G".

Analytical LCMS conditions E: residence time = 1.59 min; ESI-MS (-) m/z 898.5 (M-2H).

Analytical LCMS conditions G: residence time = 3.18 min; ESI-MS (+) m/z 899.5 (M+2H).

Preparation of Example 3226

Example 3226 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 25-65% B duration 30 Minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.2 mg and the purity was estimated to be 95% based on LCMS analysis using "analytical LCMS Condition E".

Analytical LCMS Condition E: Retention time = 1.84 min; ESI-MS (-) m/z 928.9 (M-2H).

Preparation of Example 3227

Example 3227 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 35-75% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.3 mg and the purity was estimated to be 97% based on LCMS analysis using "analytical LCMS conditions E and G".

Analytical LCMS Condition E: Retention time = 1.74 min; ESI-MS (+) m/z 947.7 (M+2H).

Analytical LCMS conditions G: residence time = 3.37 min; ESI-MS (+) m/z 948.00 (M+2H).

ESI-HRMS (+) m/z: Calculated: 149. 946 (M+2H).

Found: 949.9449 (M+2H).

Preparation of Example 3228

Example 3228 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 35-75% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product is 5.2 mg, and the root According to LCMS analysis using "analytical LCMS conditions E and G", the purity was estimated to be 98%.

Analytical LCMS conditions E: residence time = 1.80 min; ESI-MS (-) m/z 947.7 (M-2H).

Analytical LCMS conditions G: residence time = 3.41 min; ESI-MS (+) m/z 949.10 (M+2H).

ESI-HRMS (+) m/z: Calculated: 948.9357 (M+2H).

Found: 948.9354 (M+2H).

Preparation of Example 3229

Example 3229 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroethyl Acid; mobile phase B: 95:5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.6 mg and the purity was estimated to be 95% based on LCMS analysis using "analytical LCMS conditions E and G".

Analytical LCMS conditions G: retention time = 3.443 min; ESI-MS (+) m/z 965.55 (M+2H).

Preparation of Example 3230

Example 3230 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C".

The crude material was purified by preparative HPLC using the following conditions: column: Phenomenex Luna 20x250 5u particles; mobile phase A: water + 0.1% TFA; mobile phase B: acetonitrile + 0.1% TFA; gradient: 45-95% B duration 55 Minutes, then 5 minutes at 95% B; flow: 15 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. Obtained four isomers (isomers 3230-A, 3230-B, 3230-C and 3230-D). The yields of the product isomers 3230-A, 3230-B, 3230-C, and 3060-D were 7.5 mg, 9.3 mg, 0.66 mg, and 0.72 mg, respectively, and were based on "analytical HPLC Condition B" (at 60 ° C, Using a gradient of 35% to 90% buffer B for 30 minutes), the purity estimates were 97%, 97.5%, 99%, and 84%, respectively.

Analytical LCMS Condition A: Isomer 3230-A: Retention time = 1.52 min; ESI-MS (+) m/z 930.1 (M+2H).

Analytical LCMS Condition A: Isomer 3230-B: Retention time = 1.55 min ; ESI-MS (+) m/z 930.0 (M+2H).

Analytical LCMS Condition A: Isomer 3230-C: Retention time = 1.55 min ; ESI-MS (+) m/z 929.8 (M+2H).

Analytical LCMS Condition A: Isomer 3230-D: Retention time = 1.67 min; ESI-MS (+) m/z 930.2 (M+2H).

< / RTI></RTI></RTI><RTIgt;</RTI></RTI><RTIgt;

Experimental value: 9249.471 (M+2H).

< / RTI></RTI></RTI></RTI></RTI><RTIgt;

Found: 92.9372 (M+2H).

</RTI></RTI><RTIID=0.0></RTI></RTI><RTI ID= 0.0>

Found: 929.4380 (M+2H).

< / RTI></RTI></RTI></RTI></RTI></RTI><RTIgt;

Found: 929.4379 (M+2H).

Preparation of Example 3231

Example 3231 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method G" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 25-70% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.2 mg and the purity was estimated to be 99% according to LCMS analysis.

Analytical LCMS conditions D: residence time = 1.652 min; ESI-MS (+) m/z 910.40 (M+2H).

Analytical LCMS conditions E: residence time = 1.801 min; ESI-MS (+) m/z 910.75 (M+2H).

Preparation of Example 3232

Example 3232 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method G" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 20-65% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.1 mg and the purity was estimated to be 95% according to LCMS analysis.

Analytical LCMS conditions D: residence time = 1.499 min; ESI-MS (+) m/z 911.70 (M+2H).

Analytical LCMS Condition E: Retention time = 1.633 min; ESI-MS (+) m/z 911.40 (M+2H).

Preparation of Example 3233

Example 3233 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method G" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 25-70% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.3 mg and the purity was estimated to be 94% according to LCMS analysis.

Analytical LCMS conditions D: residence time = 1.662 min; ESI-MS (+) m/z 923.70 (M+2H).

Analytical LCMS Condition E: Retention time = 1.823 min; ESI-MS (+) m/z 923.75 (M+2H).

Preparation of Example 3234

Example 3234 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method G" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 25-65% B for 25 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.0 mg, and the root According to LCMS analysis, the purity was estimated to be 99%.

Analytical LCMS conditions D: residence time = 1.585 min; ESI-MS (+) m/z 922.60 (M+2H).

Analytical LCMS Condition E: Retention time = 1.724 min; ESI-MS (+) m/z 924.45 (M+2H).

Preparation of Example 3235

Example 3235 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure, "" Prelude method A: chloro acetyl chloride coupling procedure A "," general deprotection method G "and" C cyclization process. "

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 25-70% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product is 4.0 mg, and the root According to LCMS analysis, the purity was estimated to be 98%.

Analytical LCMS conditions D: retention time = 1.77 min; ESI-MS (+) m/z 923.8 (M+2H).

Analytical LCMS Condition E: Retention time = 1.99 min; ESI-MS (+) m/z 924.0 (M+2H).

Preparation of Example 3236

Example 3236 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method G" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 15-55% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.9 mg, and the root According to LCMS analysis, the purity is estimated to be 100%.

Analytical LCMS conditions D: retention time = 1.69 min; ESI-MS (+) m/z 916.3 (M+2H).

Analytical LCMS Condition E: Retention time = 1.89 min; ESI-MS (+) m/z 916.3 (M+2H).

Preparation of Example 3237

Example 3237 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method G" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.9 mg, and the root According to LCMS analysis, the purity is estimated to be 100%.

Analytical LCMS conditions D: retention time = 1.78 min; ESI-MS (+) m/z 898.4 (M+2H).

Analytical LCMS Condition E: Retention time = 1.78 min; ESI-MS (+) m/z 898.5 (M+2H).

Preparation of Example 3238

Example 3238 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method G" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.3 mg and the purity was estimated to be 91% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.78 min; ESI-MS (+) m/z s .

Analytical LCMS conditions E: residence time = 1.98 min; ESI-MS (+) m/z 902.1 (M+2H).

Preparation of Example 3239

Example 3239 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method G" and "cyclization method C". The Fmoc protected 7-aza-Trp residue is coupled as a racemate.

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 15-65% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.4 mg and the purity was estimated to be 98% as a mixture of diastereomers according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.51 min; ESI-MS (+) m/z 899.8 (M+2H).

Analytical LCMS conditions E: retention time = 1.61 min; ESI-MS (+) m/z 899.3 (M+2H).

Preparation of Example 3240

Example 3240 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method G" and "cyclization method C". The Fmoc protected 7-aza-Trp residue is coupled as a racemate.

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 15-65% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.5 mg, and its purity as a mixture of diastereomers was estimated to be 97% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.67 min; ESI-MS (+) m/z s.

Analytical LCMS Condition E: Retention time = 1.77 min; ESI-MS (+) m/z 902.5 (M+2H).

Preparation of Example 3241

Example 3241 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method G" and "cyclization method C". The Fmoc protected 7-aza-Trp residue is coupled as a racemate.

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 15-55% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.0 mg and the purity was estimated to be 98% as a mixture of diastereomers according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.66 min; ESI-MS (+) m/z 898.6 (M+2H).

Analytical LCMS Condition E: Retention time = 1.57 min; ESI-MS (+) m/z 899.2 (M+2H).

Preparation of Example 3242

Example 3242 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method G" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 15-65% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.3 mg and the purity was estimated to be 99% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.43 min; ESI-MS (+) m/z 899.5 (M+2H).

Analytical LCMS conditions E: residence time = 1.54 min; ESI-MS (+) m/z 899.6 (M+2H).

Preparation of Example 3243

Example 3243 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method G" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 15-65% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.8 mg and the purity was estimated to be 99% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.44 min; ESI-MS (+) m/z 899.5 (M+2H).

Analytical LCMS Condition E: Retention time = 1.54 min; ESI-MS (+) m/z 899.9 (M+2H).

Preparation of Example 3244

Example 3244 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method G" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 20-70% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analytical LCMS conditions D: residence time = 1.60 min; ESI-MS (+) m/z 921.3 (M+2H).

Analytical LCMS Condition E: Retention time = 1.81 min; ESI-MS (+) m/z 922.0 (M+2H).

Preparation of Example 3245

Example 3245 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method G" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 25-75% B over 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.3 mg and the purity was estimated to be 99% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.73 min; ESI-MS (+) m/z 927.1 (M+2H).

Analytical LCMS conditions E: retention time = 1.93 min; ESI-MS (+) m/z 927.4 (M+2H).

Preparation of Example 3246

Example 3246 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method G" and "cyclization method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18 300, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 15-65% B for 25 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.4 mg and the purity was estimated to be 94% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.78 min; ESI-MS (+) m/z 932.9 (M+2H).

Analytical LCMS conditions E: retention time = 1.97 min; ESI-MS (+) m/z 932.9 (M+2H).

Preparation of Example 3614

Example 3614 was prepared following the general synthetic sequence described for the preparation of Example 3210, which consisted of the following general procedure: "CEM Method A: Resin Swelling Procedure", "CEM Method A: Standard Coupling Procedure", "CEM Method A: Custom Amino Acid Coupling Procedure , "N-methylation Procedure on Resin" (for Nmethylation of Fmoc-3-PyAla-OH), " Chloroacetic Acid Coupling Procedure A ", "Total Removal Protection" Base Method B" and "Circularization Method C". Fmoc-3-PyAla-OH (2.5 eq.) was artificially coupled using HATU (2.5 eq.) and NMM (2.5 eq.) in the coupling method, followed by HATU (5 eq.) and NMM (5 eq in the coupling method). .) Fmoc-Phe-OH (5 eq.) was subjected to a second manual coupling step.

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 30-70% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.6 mg and the purity was estimated to be 94% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.53 min; ESI-MS (+) m/z 899.6 (M+2H).

Analytical LCMS conditions E: residence time = 1.45 min; ESI-MS (+) m/z 899.4 (M+2H).

ESI-HRMS (+) m/z: Calculated: 898.9345 (M+2H).

Found: 898.9345 (M+2H).

Preparation of Example 3616

Example 3616 was prepared following the general synthetic sequence described for the preparation of Example 3210, which consisted of the following general procedure: "CEM Method A: Resin Swelling Procedure", "CEM Method A: Standard Coupling Procedure", "CEM Method A: Custom Amino Acid Coupling Procedure, "N-methylation Procedure on Resin" (for Nmethylation of Fmoc-4-thiazole-ala-OH), "Chloroacetic Acid Coupling Procedure A", "Overall Detachment In addition to the protecting group method B" and "cyclization method C". Artificial coupling of Fmoc-3-(thiazol-4-yl)-Ala-OH (2.5 eq.) using HATU (2.5 eq.) and NMM (2.5 eq.) in the coupling method followed by HATU in the coupling method (5 eq.) and NMM (5 eq.) Fmoc-Phe-OH (5 eq.) was subjected to a second manual coupling step.

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-60% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The product yield is 11.8 Mg, and based on LCMS analysis, its purity was estimated to be 97%.

Analytical LCMS conditions D: retention time = 1.53 min; ESI-MS (+) m/z s .

Analytical LCMS Condition E: Retention time = 1.52 min; ESI-MS (+) m/z 902.1 (M+2H).

ESI-HRMS (+) m/z: calc.

Experimental value: 901.9130 (M+2H).

Preparation of Example 3617

Example 3617 was prepared following the general synthetic sequence described for the preparation of Example 3210, which consisted of the following general procedure: "CEM Method A: Resin Swelling Procedure", "CEM Method A: Standard Coupling Procedure", "CEM Method A : Custom Amino Acid Coupling Procedure, "Chloroethyl Chloride Chloride Coupling Procedure A", "Total Removal of Protective Group Method B" and "Circulation Method C".

The crude material was purified by preparative HPLC using the following conditions: column: Phenomenex Luna 5u C18 (2) 250 x 21.2 AXIA, 100A Ser. #520221-1; mobile phase A: 0.1% TFA/water; mobile phase B: 0.1 % TFA/acetonitrile; Gradient: 35-75% B over 40 minutes followed by a 5 minute gradient up to 85% B; flow: 15 mL/min. Consolidation The fractions of the desired product were dried and dried by centrifugal evaporation. The product was dissolved in a minimum amount of acetonitrile and water, frozen and lyophilized to afford a white amorphous solid. The yield of the product was 1.9 mg and the purity was estimated to be 90% according to LCMS analysis.

Analytical LCMS Condition A: Retention time = 1.42 min; ESI-MS (+) m/z 817.2 (M+2H).

Analytical LCMS conditions C: retention time = 1.65 min; ESI-MS (+) m/z 1632.8 (M+H).

ESI-HRMS (+) m/z: Calculated: 816.8996 (M+2H).

Found: 816.8968 (M+2H).

Preparation of Example 3618

Example 3618 was prepared following the general synthetic sequence described for the preparation of Example 3210, which consisted of the following general procedure: "CEM Method A: Resin Swelling Procedure", "CEM Method A: Standard Coupling Procedure", "CEM Method A : Custom Amino Acid Coupling Procedure, "Chloroethyl Chloride Chloride Coupling Procedure A", "Total Removal of Protective Group Method B" and "Circulation Method C".

The crude material was purified by preparative HPLC using the following conditions: column: Phenom Luna 5u C18 (2) 250 x 21.2 AXIA, 100A Ser. #520221-1; mobile phase A: 0.1% TFA/water; mobile phase B: 0.1% TFA/acetonitrile; gradient: 35-75% B over 40 minutes followed by a 5 minute gradient up to 85% B; flow: 15 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The product was dissolved in a minimum amount of acetonitrile and water, frozen and lyophilized to afford a white amorphous solid. The yield of the product was 3.5 mg and the purity was estimated to be 99% according to LCMS analysis.

Analytical LCMS Condition A: Retention time = 1.49 min; ESI-MS (+) m/z 781.7 (M+2H).

Analytical LCMS Condition C: Retention time = 1.74 min; ESI-MS (+) m/z 156 (M+H).

ESI-HRMS (+) m/z: Calculated: 781.3810 (M+2H).

Found: 781.3778 (M+2H).

Preparation of Example 3628

Example 3628 was prepared following the general synthetic sequence described for the preparation of Example 3210, which consisted of the following general procedure: "CEM Method A: Resin Swelling Procedure", "CEM Method A: Standard Coupling Procedure", "CEM Method A : Custom Amino Acid Coupling Procedure, "Chloroethyl Chloride Chloride Coupling Procedure A", "Total Removal of Protective Group Method B" and "Circulation Method C".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-60% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.97 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.45 min; ESI-MS (+) m/z 849.2 (M+2H).

Analytical LCMS Condition E: Retention time = 1.66 min; ESI-MS (+) m/z 849.3 (M+2H).

ESI-HRMS (+) m/z: Calculated: 484. 897 (M+2H).

Experimental value: 848.8962 (M+2H).

Preparation of Example 3637

Example 3637 was prepared following the general synthetic sequence described for the preparation of Example 3210, which consisted of the following general procedure: "CEM Method A: Resin Swelling Procedure", "CEM Method A: Standard Coupling Procedure", "CEM Method A: Custom Amino Acid Coupling Procedure, "N-methylation Procedure on Resin" (N-methyl for Fmoc-4-Py-ala-OH) , "Chloroacetic acid coupling procedure A", "Total removal of protecting group method B" and "Circulation method C". Fmoc-4-PyAla-OH (10 eq.) was artificially coupled using HATU (10 eq.) and NMM (20 eq.) in the coupling method, followed by HATU (5 eq.) and NMM (10 eq.) in the coupling method. Fmoc-Phe-OH (5 eq.) was subjected to a second manual coupling step.

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-60% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.78 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.74 min; ESI-MS (+) m/z 932.5 (M+2H).

Analytical LCMS Condition E: Retention time = 1.61 min; ESI-MS (+) m/z 932.5 (M+2H).

ESI-HRMS (+) m/z: calc.

Found: 931.9557 (M+2H).

Preparation of Example 3638

Example 3638 was prepared following the general synthetic sequence described for the preparation of Example 3210, which consisted of the following general procedure: "CEM Method A: Resin Swelling Procedure", "CEM Method A: Standard Coupling Procedure", "CEM Method" A: Custom Amino Acid Coupling Procedure, "N-methylation Procedure on Resin" (for N-methylation of Fmoc-4-Py-ala-OH), "Chloroacetic Acid Coupling Procedure A", "Overall Detachment In addition to the protecting group method B" and "cyclization method C". Fmoc-4-PyAla-OH (10 eq.) was artificially coupled using HATU (10 eq.) and NMM (20 eq.) in the coupling method, followed by HATU (5 eq.) and NMM (10 eq.) in the coupling method. Fmoc-Phe-OH (5 eq.) was subjected to a second manual coupling step.

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-60% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. Separation produces two isomers. The yield of the first isomer 3638-A was 1.66 mg and its purity was estimated to be 91% according to LCMS analysis.

Analytical LCMS conditions D: residence time = 1.07 min; ESI-MS (+) m/z 899.5 (M+2H).

Analytical LCMS conditions E: residence time = 1.08 min; ESI-MS (+) m/z 899.4 (M+2H).

The yield of the second isomer 3638-B was 7.56 mg and the purity was estimated to be 92% according to LCMS analysis.

Analytical LCMS conditions D: retention time = 1.52 min; ESI-MS (+) m/z 899.2 (M+2H).

Analytical LCMS Condition E: Retention time = 1.45 min; ESI-MS (+) m/z 899.5 (M+2H).

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Found: 898.9331 (M+2H).

Preparation of Example 3639

Example 3639 was prepared following the general synthetic sequence described for the preparation of Example 3210, which consisted of the following general procedure: "CEM Method A: Resin Swelling Procedure", "CEM Method A: Standard Coupling Procedure", "CEM Method A : Custom Amino Acid Coupling Procedure, "Chloroethyl Chloride Chloride Coupling Procedure A", "Total Removal of Protective Group Method B" and "Circulation Method C".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-60% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product as a mixture of diastereomers were combined and dried by centrifugal evaporation. The yield of the product was 6.7 mg and the purity was estimated to be 96% according to LCMS analysis.

Analytical LCMS conditions D: residence time = 1.65 min; ESI-MS (+) m/z 903.45 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Found: 905.4376 (M+2H).

Preparation of Example 3640

The two diastereomers from Example 3639 were separated by supercritical fluid chromatography (SFC) on a Berger SFC MGII system using the following conditions: column: ES DEAP 25 x 2.1 cm 5 μm particles; mobile phase : 44/55 CO ₂ /95:5 MeOH: H ₂ O + 10 mM NH ₄ OAc.

The yield of the first isomer 3640-A was 2.39 mg and its purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions A: residence time = 1.43 min; ESI-MS (+) m/z 1812.1 (M+H).

Analytical LCMS conditions C: retention time = 1.67 min; ESI-MS (+) m/z 1810.2 (M+H).

The yield of the second isomer 3640-B was 2.15 mg and its purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS Condition A: Retention time = 1.42 min; ESI-MS (+) m/z 1812.1 (M+H).

Analytical LCMS conditions C: retention time = 1.66 min; ESI-MS (+) m/z 1811.1 (M+H).

Preparation of Example 3641

Example 3641 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C". Fmoc-Val-OH was coupled to the peptidyl resin using 10 eq. amino acid, 10 eq. HATU and 20 eq. NMM and extended to 10 hours. 1.5 eq. Amino acid, 1.65 eq. 7-aza-benzotriazole (HOAt) and 1.58 eq. N,N-diisopropylcarbonyldiimide (DIC) were used in DMF. The final coupling of Fmoc-Phe(3,4,5-tri-F)-OH was preformed artificially and allowed to proceed for 60 hours.

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 Methanol: water + 10 mM ammonium acetate; gradient: 40-90% B It lasted for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.9 mg and the purity was estimated to be 100% based on LCMS analysis using "analytical LCMS conditions D and G".

Analytical LCMS conditions D: retention time = 1.61 min; ESI-MS (-) m/z 948.7 (M-2H).

Analytical LCMS conditions G: residence time = 2.98 min; ESI-MS (+) m/z 950.7 (M+2H).

ESI-HRMS(+) m/z: Calculated value: (M+2H)949.9253

Experimental value: (M+2H) 949.9230

Preparation of Example 3642

Example 3642 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C". Fmoc-Val-OH was coupled to the peptidyl resin using 10 eq. amino acid, 10 eq. HATU and 20 eq. NMM and extended to 10 hours. 1.5 eq. Amino acid, 1.65 eq. 7-aza-benzotriazole (HOAt) and 1.58 eq. N,N-diisopropylcarbonyldiimide (DIC) were used in DMF. The final coupling of Fmoc-Phe(3,4,5-tri-F)-OH was preformed artificially and allowed to proceed for 60 hours.

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 Methanol: water + 10 mM ammonium acetate; gradient: 40-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.8 mg and the purity was estimated to be 100% based on LCMS analysis using "analytical LCMS conditions D and G".

Analytical LCMS conditions D: residence time = 1.63 min; ESI-MS (+) m/z 950.8 (M-2H).

Analytical LCMS conditions G: residence time = 2.98 min; ESI-MS (+) m/z 950.8 (M+2H).

ESI-HRMS(+) m/z: Calculated value: (M+2H)949.9253

Experimental value: (M+2H) 949.9229

Preparation of Example 3643

Example 3643 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C". Fmoc-Val-OH was coupled to the peptidyl resin using 10 eq. amino acid, 10 eq. HATU and 20 eq. NMM and extended to 10 hours. 1.5 eq. Amino acid, 1.65 eq. 7-aza-benzotriazole (HOAt) and 1.58 eq. N,N-diisopropylcarbonyldiimide (DIC) were used in DMF. The final coupling of Fmoc-Phe(3,4,5-tri-F)-OH was preformed artificially and allowed to proceed for 60 hours.

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 Methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 31.8 mg and the purity was estimated to be 98% based on LCMS analysis using "analytical LCMS conditions D and G".

Analytical LCMS conditions D: retention time = 1.52 min; ESI-MS (+) m/z 936.9 (M+2H).

Analytical LCMS conditions G: retention time = 2.90 min; ESI-MS (+) m/z 936.8 (M+2H).

ESI-HRMS(+) m/z: calculated: (M+2H) 935.9097

Experimental value: (M+2H) 935.9073.

Preparation of Example 3644

Example 3644 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C". Fmoc-Val-OH was coupled to the peptidyl resin using 10 eq. amino acid, 10 eq. HATU and 20 eq. NMM and extended to 10 hours. 1.5 eq. Amino acid, 1.65 eq. 7-aza-benzotriazole (HOAt) and 1.58 eq. N,N-diisopropylcarbonyldiimide (DIC) were used in DMF. The final coupling of Fmoc-Phe(3,4,5-tri-F)-OH was preformed artificially and allowed to proceed for 60 hours.

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 Methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 35.9 mg and the purity was estimated to be 100% based on LCMS analysis using "analytical LCMS conditions D and G".

Analytical LCMS conditions D: retention time = 1.57 min; ESI-MS (+) m/z 936.5 (M+2H).

Analytical LCMS conditions G: residence time = 2.93 min; ESI-MS (+) m/z 936.5 (M+2H).

ESI-HRMS(+) m/z: calculated: (M+2H) 935.9097

Experimental value: (M+2H) 935.9069.

Preparation of Example 3645

Example 3645 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C". Fmoc-Val-OH was coupled to the peptidyl resin using 10 eq. amino acid, 10 eq. HATU and 20 eq. NMM and extended to 10 hours. 1.5 eq. Amino acid, 1.65 eq. 7-aza-benzotriazole (HOAt) and 1.58 eq. N,N-diisopropylcarbonyldiimide (DIC) were used in DMF. The final coupling of Fmoc-Phe(3,4,5-tri-F)-OH was preformed artificially and allowed to proceed for 60 hours.

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 Methanol: water + 10 mM ammonium acetate; gradient: 50-100% B lasted for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 46.3 mg and the purity was estimated to be 95% based on LCMS analysis using "analytical LCMS conditions D and G".

Analytical LCMS conditions D: residence time = 1.82 min; ESI-MS (-) m/z 941.05 (M-2H).

Analytical LCMS conditions G: residence time = 3.06 min; ESI-MS (+) m/z 943.8 (M+2H).

ESI-HRMS(+) m/z: Calculated: (M+2H) 942.9175

Found: (M+2H) 942.9159.

Preparation of Example 3646

Example 3646 was prepared following the general synthetic sequence described for the preparation of Example 3212, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", "Prelude Method" A: secondary amine coupling procedure , "Prelude method A: chloroacetic chloride coupling procedure A", "total removal of protecting group method B" and "cyclization method C". Fmoc-Val-OH was coupled to the peptidyl resin using 10 eq. amino acid, 10 eq. HATU and 20 eq. NMM and extended to 10 hours. 1.5 eq. Amino acid, 1.65 eq. 7-aza-benzotriazole (HOAt) and 1.58 eq. N,N-diisopropylcarbonyldiimide (DIC) were used in DMF. The final coupling of Fmoc-Phe(3,4,5-tri-F)-OH was preformed artificially and allowed to proceed for 60 hours.

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 250 mm, 5 μm particles: mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 Methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.3 mg and the purity was estimated to be 98% based on LCMS analysis using "analytical LCMS conditions D and G".

Analytical LCMS conditions D: retention time = 1.57 min; ESI-MS (+) m/z 943.4 (M+2H).

Analytical LCMS conditions G: residence time = 2.93 min; ESI-MS (+) m/z 94 </RTI> (M+2H).

ESI-HRMS(+) m/z: Calculated: (M+2H) 942.9175

Found: (M+2H) 942.9152.

Preparation of Example 3647

Sieber resin (140 mg, 0.100 mmol) was added to a Symphony polypropylene solid phase reaction vessel, and the reaction vessel was placed on a Symphony peptide synthesizer. Then execute the following procedures in sequence: use, follow "Symphony Method A: Resin Swelling Procedure"; use Fmoc-Gly-OH, follow "Symphony Method A: Standard Coupling Procedure"; use Fmoc-Cys(Trt)-OH, follow "Symphony" Method A: Standard coupling procedure"; use Fmoc-Leu-OH, follow "Symphony Method A: Standard Coupling Procedure"; use Fmoc-Tyr(tBu)-OH, follow "Symphony Method A: Standard Coupling Procedure"; use Fmoc- Trp(Boc)-OH, follow "Symphony Method A: Standard Coupling Procedure"; use Fmoc-(D)-morpholinyl-3-carboxylic acid, follow "Symphony Method A: Standard Coupling Procedure"; use Fmoc-Glu (OtBu) )-OH, follow "Symphony Method A: Secondary Amine Coupling Procedure "; use Fmoc-[N-Me]Phe-OH, follow "Symphony Method A: Standard Coupling Procedure"; use Fmoc-Val-OH, follow "Symphony" Method A: Secondary amine coupling procedure "; using Fmoc-Asp(OtBu)-OH, following "Symphony Method A: Single Standard Coupling Procedure"; using Fmoc-(D)-cis-Pro(4-OH)-OH, Follow "Symphony Method A: Single Standard Coupling Procedure"; use Fmoc-[N-Me]Nle-OH, follow the "Prelude side" Method A: Second amine coupling procedure "; Use Fmoc-[N-Me]Phe-OH, follow "Prelude method A: secondary amine coupling procedure "; use Fmoc-Phe(3-Cl)-OH, follow "Prelude Method A: Secondary amine coupling procedure "; use, follow "Symphony Method B: Final Ending Procedure "; use, follow "Total Removal Protection Method B"; use, follow "Circularization Method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions D: residence time = 1.73 min; ESI-MS (-) m/z 939.8 (M-2H).

Analytical LCMS conditions G: residence time = 2.9 min; ESI-MS (+) m/z 941.8 (M+2H).

ESI-HRMS(+) m/z: calculated: (M+2H) 940.9096

Experimental value: (M+2H) 940.9063.

Preparation of Example 3648

Example 3648 was prepared on Rink resin following the general synthetic procedure described for the preparation of Example 3647. The general synthetic sequence consisted of the following general procedure: "Symphony Method A: Resin Swelling Procedure", "Symphony Method A: Standard Coupling Procedure" "Symphony Method A: Secondary Amine Coupling Procedure", "Symphony Method B: Final Ending Procedure", "Total Removal of Protective Base Method B" and "Circularization Method C".

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical LCMS conditions D: residence time = 1.67 min; ESI-MS (-) m/z 931.1 (M-2H).

Analytical LCMS conditions G: residence time = 2.84 min; ESI-MS (+) m/z 933.3 (M+2H).

ESI-HRMS(+) m/z: Calculated: (M+2H) 932.9244

Experimental value: (M+2H) 932.9200.

Analysis data:

Mass spectrometry: "ESI-MS (+)" indicates electrospray ionization mass spectrometry in positive ion mode; "ESI-MS (-)" indicates electrospray ionization mass spectrometry in negative ion mode; "ESI-HRMS (+)" indicates High-resolution electrospray ionization mass spectrometry in positive ion mode; "ESI-HRMS(-)" represents high-resolution electrospray ionization mass spectrometry in negative ion mode. The detected quality is reported after the " m/z " unit name. Compounds with an accurate mass greater than 1000 are often detected as ions with double or triple charges.

Analysis condition A:

Column: Waters BEH C18, 2.0×50 mm, 1.7 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; temperature: 50 ° C; Gradient: 0% B, 0-100% B lasted 3 minutes, then held at 100% B for 0.5 minutes; flow rate: 1 mL/min; detection: 220 nm UV.

Analysis condition B:

Column: Waters BEH C18, 2.0×50 mm, 1.7 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; temperature: 50 ° C; Gradient: 0% B, 0-100% B lasted 3 minutes, then held at 100% B for 0.5 minutes; flow rate: 0.5 mL/min; detection: 220 nm UV.

Analysis condition C:

Column: Waters Aquity BEH C18 2.1 × 50 mm 1.7 μm particles; mobile phase A: water + 0.05% TFA; mobile phase B: acetonitrile + 0.05% TFA; temperature: 40 ° C; gradient: 0% B, 0-100% B It lasted for 3 minutes, then held at 100% B for 0.5 minutes; flow rate: 0.8 mL/min; detection: 220 nm UV.

Analysis condition D:

Column: Waters Aquity BEH C18 2.1×50mm 1.7μm particles; mobile phase A: water +0.05% TFA; mobile phase B: methanol +0.05% TFA; temperature: 40°C; ladder Degree: 0% B, 0-100% B lasted 3 minutes, then held at 100% B for 0.5 minutes; flow rate: 0.8 mL/min; detection: 220 nm UV.

General procedure: Prelude Method A:

All operations were performed on a Prelude Peptide Synthesizer (Protein Technologies) according to automated operation. Unless otherwise noted, all procedures were carried out in a 10 mL polypropylene tube equipped with a bottom frit; in the case of a reaction scale exceeding 0.100 mmol, a 40 mL polypropylene tube fitted with a bottom frit was used. The tube is connected to the Prelude Peptide Synthesizer via the bottom and top of the tube. DMF and DCM can be added via the top of the tube, which is equally flushed along the side of the tube. The remaining reagent is added via the bottom of the tube and passed up through the frit to contact the resin. All solutions were removed via the bottom of the tube. "Stirred periodically" described in brief pulses N ₂ gas through the bottom frit; pulse duration from about 5 seconds and occur once every 30 seconds. The solution of chloroacetic chloride in DMF was used within 24 hours of preparation. The amino acid solution is usually used up to three weeks after preparation. The HATU solution was used within 5 days of preparation. DMF = dimethylformamide; HATU = hexafluorophosphate 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3 -Oxide; DIPEA = diisopropylethylamine; Rink = (2,4-dimethoxyphenyl)(4-alkoxyphenyl)methylamine, wherein "4-alkoxy" is described and described The location and type of polystyrene resin connections. The resin used was Merrifield polymer (polystyrene) with Rink linker (nitrogen protected by Fmoc); 100-200 mesh, 1% DVB, 0.56 mmol/g loading. The common amino acids used are listed below, wherein the side chain protecting groups are indicated in parentheses.

Fmoc-Ala-OH; Fmoc-Arg(Pbf)-OH; Fmoc-Asn(Trt)-OH; Fmoc-Asp(OtBu)-OH; Fmoc-Bzt-OH; Fmoc-Cys(Trt)-OH; Fmoc- Dab(Boc)-OH; Fmoc-Dap(Boc)-OH; Fmoc-Gln(Trt)-OH; Fmoc-Gly-OH; Fmoc-His(Trt)-OH; Fmoc-Hyp(tBu)-OH; Fmoc -Ile-OH; Fmoc-Leu-OH; Fmoc-Lys(Boc)-OH; Fmoc-Nle-OH; Fmoc-Met-OH; Fmoc-[N-Me]Ala-OH; Fmoc-[N-Me] Nle-OH; Fmoc-Phe-OH; Fmoc-Pro-OH; Fmoc-Sar-OH; Fmoc-Ser(tBu)-OH; Fmoc-Thr(tBu)-OH; Fmoc-Trp(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Val-OH.

The procedure of "Prelude Method A" describes an experiment conducted on a scale of 0.100 mmol, where the scale is determined by the amount of Rink linker bound to the resin. This scale corresponds to about 178 mg of the above Rink-Merrifield resin. All procedures can be scaled up by more than 0.100 mmol by adjusting the volume by a multiple of this size. Prior to amino acid coupling, all peptide synthesis sequences begin with a resin swelling procedure, described below as "resin swelling procedure." The N-terminal coupling of an amino acid with a primary amine uses the following "single coupling procedure". The N-terminal coupling of the amino acid with the secondary amine uses the following "double coupling procedure". The coupling of chloroacetamidine chloride to the N-terminus of the peptide is described by the "chloroethonium chloride coupling procedure" detailed below.

Resin swelling procedure:

Merrifield: Rink resin (178 mg, 0.100 mmol) was added to a 10 mL polypropylene solid phase reaction vessel. The resin was washed (swelled) three times as follows: DMF (2.0 mL) was added to the reaction vessel, and then the mixture was periodically stirred for 10 minutes, and then the solvent was discharged through a glass frit.

Single coupling procedure:

Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to a reaction vessel containing the resin from the previous step. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. The resin was washed six times in succession as follows: DMF (2.0 mL) was added via the top of the vessel for each wash and the resulting mixture was stirred periodically for 30 seconds before the solution was discharged through the frit. Amino acid (0.2 M in DMF, 1.0 mL, 2 eq) was added to the reaction vessel, followed by HATU (0.2M in DMF, 1.0 mL, 2 eq), and finally DIPEA (0.8 M in DMF, 0.5 mL) , 4eq). The mixture was stirred periodically for 15 minutes, and then the reaction solution was discharged through a frit. The resin was washed four times as follows: DMF (2.0) was added via the top of the vessel for each wash. (mL) and the resulting mixture was periodically stirred for 30 seconds, after which the solution was discharged through a frit. Acetic anhydride (2.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 10 minutes and then the solution was discharged through a frit. The resin was washed four times continuously as follows: DFI (2.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 90 seconds, after which the solution was discharged through the frit. The resulting resin was used directly in the next step.

Double coupling procedure

Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to a reaction vessel containing the resin from the previous step. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. The resin was washed six times in succession as follows: DMF (2.0 mL) was added via the top of the vessel for each wash and the resulting mixture was stirred periodically for 30 seconds before the solution was discharged through the frit. Amino acid (0.2 M in DMF, 1.0 mL, 2 eq) was added to the reaction vessel, followed by HATU (0.2M in DMF, 1.0 mL, 2 eq), and finally DIPEA (0.8 M in DMF, 0.5 mL) , 4eq). The mixture was stirred periodically for 15 minutes, and then the reaction solution was discharged through a frit. The resin was washed twice as follows: at each wash, DMF (2.0 mL) was added via the top of the vessel and the resulting mixture was stirred periodically for 30 seconds, then the solution was drained through the frit. Amino acid (0.2 M in DMF, 1.0 mL, 2 eq) was added to the reaction vessel, followed by HATU (0.2M in DMF, 1.0 mL, 2 eq), and finally DIPEA (0.8 M in DMF, 0.5 mL) , 4eq). The mixture was stirred periodically for 15 minutes, and then the reaction solution was discharged through a frit. The resin was washed twice as follows: at each wash, DMF (2.0 mL) was added via the top of the vessel and the resulting mixture was stirred periodically for 30 seconds, then the solution was drained through the frit. Acetic anhydride (2.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 10 minutes and then the solution was discharged through a frit. The resin was washed four times continuously as follows: DFI (2.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 90 seconds, after which the solution was discharged through the frit. The resulting resin was used directly in the next step.

Chloroacetyl chloride coupling procedure:

Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to a reaction vessel containing the resin from the previous step. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. The resin was washed six times in succession as follows: DMF (2.0 mL) was added via the top of the vessel for each wash and the resulting mixture was stirred periodically for 30 seconds before the solution was discharged through the frit. DIPEA (0.8 M in DMF, 3.0 mL, 24 eq) was added to the reaction mixture, followed by chloroethyl chloride (0.8M in DMF, 1.65 mL, 13.2 eq). The mixture was stirred periodically for 30 minutes, and then the reaction solution was discharged through a glass frit. The resin was washed three times in succession as follows: DFI (2.0 mL) was added to the vessel at the top of each wash and the resulting mixture was periodically stirred for 90 seconds, after which the solution was discharged through the frit. The resin was washed four times in the following manner: On each wash, CH ₂ Cl ₂ (2.0 mL) was added to the vessel at the top and the resulting mixture was stirred periodically for 90 seconds, after which the solution was discharged through the glass frit. The resulting resin was placed under a stream of N ₂ for 15 minutes.

Symphony Method A:

This collection procedure is the same as "Prelude Method A" unless otherwise noted. In all procedures, a Prelude Peptide Synthesizer was replaced with a Symphony X Peptide Synthesizer (Protein Technologies) and all reagents were added via the top of the reaction vessel.

Resin swelling procedure:

This procedure is the same as "Prelude Method A: Resin Swelling Procedure".

Single coupling procedure:

This procedure was identical to "Prelude Method A: Single Coupling Procedure", but with a DIPEA solution concentration of 0.4 M and 1.0 mL of this solution was delivered to the reaction.

Double coupling procedure:

This procedure was identical to the "Prelude Method A: Double Coupling Procedure", but with a DIPEA solution concentration of 0.4 M and 1.0 mL of this solution was delivered to the reaction.

Chloroacetyl chloride coupling procedure:

This procedure is the same as "Prelude Method A: Chlorochlorohydrin Chloride Coupling Procedure".

Overall removal of the protective method A:

All operations are performed manually unless otherwise noted. The procedure for "Total Removal of Protecting Group Method A" describes an experiment conducted on a scale of 0.100 mmol, wherein the scale is determined by the amount of Rink linker bound to the resin. The procedure can be expanded by more than 0.100 mmol by adjusting the volume in multiples of this size. The "removal of the protecting group solution" was prepared by combining trifluoroacetic acid (22 mL), phenol (1.325 g), water (1.25 mL), and triisopropyl decane (0.5 mL) in a 40 mL glass vial. The resin was removed from the reaction vessel and transferred to a 4 mL glass vial. A "Removal of the protecting group solution" (2.0 mL) was added to the vial. The mixture was vigorously mixed in an oscillator (1000 RPM for 1 minute followed by 500 RPM for 1 to 2 hours). The mixture was filtered through a 0.2 micron syringe filter and the solid was extracted with a &lt;RTI ID=0.0&gt;&gt; To a 24 mL tube containing the combined filtrate was added Et ₂ O (15 mL). The mixture was vigorously mixed, followed by precipitation of a large amount of white solid. The mixture was centrifuged for 5 minutes, then the solution was decanted from the solid and discarded. The solid was suspended in Et ₂ O (20 mL); the mixture was then centrifuged for 5 min; and the solution was decanted from the solid and discarded. Finally, the solid was suspended in Et ₂ O (20 mL); the mixture was centrifuged for 5 minutes; and the solution was decanted from the solid and discarded to give a crude peptide as a white to pale white solid.

Cycling method A:

All operations are performed manually unless otherwise noted. The procedure of "Catalylation Method A" describes an experiment conducted on a scale of 0.100 mmol, wherein the scale is determined by the amount of Rink linker bound to the resin used to produce the peptide. This scale is not directly determined by the amount of peptide used in the program. The procedure can be expanded by more than 0.100 mmol by adjusting the volume in multiples of this size. The crude solid was dissolved peptide MeCN: 0.1M NH ₄ OAc aq (1: 1) until a total volume of 18 to 22mL, and then the solution was carefully adjusted to pH = 8.5-9.0 with aqueous NaOH (1.0M). The solution was then allowed to stand for 12 to 18 hours without agitation. The reaction solution was concentrated and the residue was dissolved in DMSO:MeOH. This solution was subjected to reverse phase HPLC purification to give the desired cyclic peptide.

General synthetic sequence A:

"General Synthesis Sequence A" describes the general sequence of procedures used to obtain the cyclic peptides described herein. For the purpose of this general procedure, the program "Symphony Method A" can be interchanged with "Prelude Method A". Rink-Merrifield resin (178 mg, 0.100 mmol) was added to a 10 mL polypropylene solid phase reaction vessel, and the reaction vessel was placed on a Prelude peptide synthesizer. Follow "Prelude Method A: Resin Swelling Procedure". Next, on Prelude, follow the "Prelude Method A: Single Coupling Procedure" (if the N-terminus of the resin-binding peptide is a primary amine) or "Prelude Method A: Double Coupling Procedure" (if the N-terminus of the resin-binding peptide is a secondary amine) , a series of amino acid couplings are carried out in sequence. Follow the "Prelude Method A: Chlorochlorohydrin Chloride Coupling Procedure"; then follow the "Total Removal of Protecting Method A"; then follow the "Circularization Method A".

General Synthetic Sequence B:

"General Synthesis Sequence B" describes the general sequence of procedures used to obtain the cyclic peptides described herein. For the purpose of this general procedure, the program "Symphony Method A" can be interchanged with "Prelude Method A". To a 10 mL polypropylene solid phase reaction vessel was added 2-chlorotrityl chloride resin (100 mg, 1.42 mmol/g loading). The reaction vessel was placed on a Prelude peptide synthesizer. A solution of the C-terminal Fmoc protected amino acid (0.10 mmol) and diisopropylethylamine (0.65 mmol) in DCM (2.5 mL) was added. The mixture was stirred for 60 minutes by periodic nitrogen bubbling under automatic operation. Methanol (0.20 mL) was added to the reaction vessel. The mixture was stirred by periodic nitrogen bubbling for 15 minutes and then discharged through a frit. The resin was washed three times in succession as follows: DCM (2.0 mL) was added to the vessel at the top of each wash and the resulting mixture was stirred periodically for 90 seconds before the solution was discharged through the frit. The resin was washed three times in succession: at each wash, DMF (2.0 mL) was added to the vessel at the top and the resulting mixture was stirred periodically for 90 seconds, then the solution was passed Exhaust through the frit. The resin was washed twice in succession as follows: DCM (2.0 mL) was added to the vessel at the top of each wash and the resulting mixture was stirred periodically for 90 seconds before the solution was discharged through the frit. The resin was washed three times in succession as follows: DFI (2.0 mL) was added to the vessel at the top of each wash and the resulting mixture was periodically stirred for 90 seconds, after which the solution was discharged through the frit. Next, on Prelude, follow the "Prelude Method A: Single Coupling Procedure" (if the N-terminus of the resin-binding peptide is a primary amine) or "Prelude Method A: Double Coupling Procedure" (if the N-terminus of the resin-binding peptide is a secondary amine) , a series of amino acid couplings are carried out in sequence. Follow the "Prelude Method A: Chlorochlorohydrin Chloride Coupling Procedure"; then follow the "Total Removal of Protecting Method A"; then follow the "Circularization Method A".

Preparation of (S)-2-(((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(1-(2-(t-butoxy)-2-yloxy) Ethyl)-1H-indol-3-yl)propionic acid

step 1:

To (S)-2-(((benzyloxy)carbonyl)amino)-3-(1H-indol-3-yl)propanoic acid benzyl ester (25.0 g, 58.3 mmol) and cesium carbonate (20.9) g, 64.2 mmol) 2-bromoacetic acid tert-butyl ester (9.36 mL, 64.2 mmol) was added to a solution of EtOAc (EtOAc). The solution was slowly warmed to room temperature with stirring for 18 hours. The reaction mixture was poured into ice water: 1N aq. The organic layer was washed with brine, collected, dried over MgSO _4, filtered and then concentrated in vacuo. The resulting solid was subjected to SiO ₂ chromatography (330 g column, 0-50% EtOAc:Hex, 20 column volume) to afford 29.6 g (93%) of (S)-2- (( Methoxy)carbonyl)amino)-3-(1-(2-(t-butoxy)-2-oxooxyethyl)-1H-indol-3-yl)propionic acid benzyl ester.

Step 2:

H ₂ was slowly bubbled through (S)-2-(((benzyloxy)carbonyl)amino)-3-(1-(2-(t-butoxy)-2-) at room temperature a mixture of phenyloxyethyl)-1H-indol-3-yl)propionate (29.6 g, 54.5 mmol) and Pd-C (1.45 g, 1.36 mmol) in MeOH (200 mL) . The mixture was then stirred under a positive pressure of H _2, while the conversion rate monitored by LCMS. After 48 hours, it was judged that the conversion was completed and the reaction mixture was filtered through celite and evaporated to give crude (S)-2-amino-3-(1-(2-(t-butoxy)-2- ox. Base ethyl)-1H-indol-3-yl)propionic acid (17.0 g) was transferred to Step 3 without additional purification.

Step 3:

To (S)-2-amino-3-(1-(2-(t-butoxy)-2-oxoethyl)-1H-indol-3-yl)propanoic acid (5.17 g, 16.2 mmol) and sodium bicarbonate (6.8 g, 81 mmol) in a solution of acetone: water (50.0 ml: 100 ml) (2,5-di-oxypyrrolidin-1-yl)carbonate (9H-indole- 9-yl)methyl ester (5.48 g, 16.2 mmol). The mixture was stirred overnight and then LCMS analysis indicated complete conversion. The vigorously stirred mixture was acidified by the slow addition of 1 N aqueous HCl. The mixture was diluted with DCM (150 mL) and the separated organic phase was washed with water and then brine. The organic layer was collected, dried over sodium sulfate and evaporated The crude material was purified by EtOAc (EtOAc EtOAc (EtOAc:EtOAc) -茀-9-yl)methoxy)carbonyl)amino)-3-(1-(2-(t-butoxy)-2-yloxyethyl)-1H-indol-3-yl ) Propionic acid. ¹ H NMR (500 MHz, methanol-d ₄ ) d 7.80 (d, J = 7.6 Hz, 2H), 7.67-7.60 (m, 2H), 7.39 (t, J = 7.5 Hz, 2H), 7.32-7.22 (m) , 3H), 7.18 (td, J = 7.6, 0.9 Hz, 1H), 7.08 (td, J = 7.5, 0.9 Hz, 1H), 7.04 (s, 1H), 4.54 (dd, J = 8.4, 4.9 Hz, 1H), 4.36-4.23 (m, 2H), 4.23-4.14 (m, 1H), 3.43-3.35 (m, 2H), 3.25-3.09 (m, 1H), 1.55-1.38 (m, 9H). ESI- MS (+) m / z = 541.3 (M + H).

Preparation of Example 5148

Example 5148 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.57 min; ESI-MS (+) m/z 972.2 (M+2H).

Analysis conditions B: residence time = 2.76 min; ESI-MS (+) m/z 972.3 (M+2H).

ESI-HRMS (+) m/z: Calculated: 97.

Preparation of Example 5150

Example 5150 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 200 mm, 5 Mm particles; mobile phase A: 5:95 methanol: water + 0.1% trifluoroacetic acid; mobile phase B: 95:5 methanol: water + 0.1% trifluoroacetic acid; gradient: 35-75% B lasts 30 minutes, then 100% B for 5 minutes; flow rate: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.36 min; ESI-MS (-) m/z 948.7 (M-2H).

Analysis condition B: residence time = 2.46 min; ESI-MS (-) m/z 948.3 (M-2H).

ESI-HRMS (+) m/z: Calculated: 495.

Preparation of Example 5151

Example 5151 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. Combine the fractions containing the desired product and steam by centrifugation Hair is dried. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 1.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.41 min; ESI-MS (+) m/z 1833.3 (M+H) analysis condition B: retention time = 2.55 min; ESI-MS (-) m/z 915.7 (M-2H) .

ESI-HRMS (+) m/z: Calculated: 916.9374.

Preparation of Example 5152

Example 5152 was prepared following "General Synthetic Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol :water+ 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 32.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.13 min; ESI-MS (+) m/z 943.6 (M+2H).

Analysis conditions B: residence time = 2.23 min; ESI-MS (+) m/z 943.6 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5153

Example 5153 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile :water +10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 49.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.458 min; ESI-MS (+) m/z 953.15 (M+2H).

Analysis condition B: residence time = 2.83 min; ESI-MS (-) m/z 950.30 (M-2H).

ESI-HRMS (+) m/z: Calculated: 195.

Preparation of Example 5154

Example 5154 was prepared following "General Synthetic Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. Combine the fractions containing the desired product and evaporate by centrifugation Dry it. The yield of the product was 20.7 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.523 min; ESI-MS (+) m/z 946.40 (M+2H).

Analysis conditions B: residence time = 2.738 min; ESI-MS (+) m/z 946.10 (M+2H).

ESI-HRMS (+) m/z: Calculated: 945.922.

Preparation of Example 5155

Example 5155 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. Combine the fractions containing the desired product and steam by centrifugation Hair is dried. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 20 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.4 mg and the purity was estimated to be 94% according to LCMS analysis.

Analysis conditions A: residence time = 1.42 min; ESI-MS (+) m/z 978.1 (M+2H).

Analysis conditions B: residence time = 2.61 min; ESI-MS (+) m/z 977.7 (M+2H).

ESI-HRMS (+) m/z: Calculated: 977.4.

Preparation of Example 5156

Example 5156 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19×mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol :Water +10 mM ammonium acetate; gradient: 30-75% B for 30 minutes followed by 100 minutes at 100% B; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.5 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.23 min; ESI-MS (+) m/z 97.6 (M+2H).

Analysis condition B: residence time = 2.35 min; ESI-MS (-) m/z 969.5 (M-2H).

ESI-HRMS (+) m/z: Calculated: 97.

Preparation of Example 5157

Example 5157 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19×mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. Combine the fractions containing the desired product and evaporate by centrifugation Dry it. The yield of the product was 22.2 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.35 min; ESI-MS (+) m/z 929.7 (M+2H).

Analysis condition B: residence time = 2.50 min; ESI-MS (+) m/z 930.3 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5158

Example 5158 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 0.1% trifluoroacetic acid; mobile phase B : 95:5 methanol: water + 0.1% trifluoroacetic acid; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.25 min; ESI-MS (+) m/z 972.1 (M+2H).

Analysis conditions B: residence time = 2.39 min; ESI-MS (+) m/z 971.6 (M+2H).

ESI-HRMS (+) m/z: Calculated: 97.

Preparation of Example 5159

Example 5159 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.9 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z 936.5 (M+2H).

Analysis conditions B: residence time = 2.77 min; ESI-MS (+) m/z 936.3 (M+2H).

ESI-HRMS (+) m/z: Calculated: 303.

Preparation of Example 5160

Example 5160 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.0 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.36 min; ESI-MS (+) m/z 972.1 (M+2H).

Analysis conditions B: residence time = 2.46 min; ESI-MS (+) m/z 971.8 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 5161

Example 5161 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.7 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.44 min; ESI-MS (+) m/z 923.2 (M+2H).

Analysis condition B: residence time = 2.60 min; ESI-MS (-) m/z 921.6 (M-2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5162

Example 5162 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.2 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.47 min; ESI-MS (+) m/z 923.8 (M+2H).

Analysis conditions B: residence time = 2.63 min; ESI-MS (+) m/z 923.8 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5163

Example 5163 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 25-45% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-35% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 49.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.28 min; ESI-MS (+) m/z 958.7 (M+2H).

Analysis conditions B: residence time = 2.41 min; ESI-MS (+) m/z 957.8 (M+2H).

ESI-HRMS (+) m/z: Calculated: 95.

Preparation of Example 5164

Example 5164 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.36 min; ESI-MS (-) m/z 915.3 (M-2H).

Analysis conditions B: residence time = 2.54 min; ESI-MS (+) m/z 917.6 (M+2H).

ESI-HRMS (+) m/z: Calculated: 916.9374

Preparation of Example 5165

Example 5165 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95 : 5 methanol: water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.45 min; ESI-MS (-) m/z 914.8 (M-2H).

Analysis condition B: residence time = 2.58 min; ESI-MS (+) m/z 917.0 (M+2H).

ESI-HRMS (+) m/z: Calculated: 916.4454.

Preparation of Example 5166

Example 5166 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 20 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.41 min; ESI-MS (+) m/z 923.0 (M+2H).

Analysis condition B: residence time = 2.58 min; ESI-MS (+) m/z 923.1 (M+2H).

Preparation of Example 5167

Example 5167 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 37.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.38 min; ESI-MS (+) m/z 931.8 (M+2H).

Analysis conditions B: residence time = 2.54 min; ESI-MS (+) m/z 931.3 (M+2H).

ESI-HRMS (+) m/z: Calculated: 93.

Preparation of Example 5168

Example 5168 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.374 min; ESI-MS (+) m/z 931.4 (M+2H).

Analysis condition B: residence time = 2.892 min; ESI-MS (+) m/z 931.4 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5169

Example 5169 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.538 min; ESI-MS (+) m/z 940.60 (M+2H).

Analysis condition B: residence time = 2.832 min; ESI-MS (-) m/z 940.65 (M-2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5170

Example 5170 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.2 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.36 min; ESI-MS (+) m/z 1849.6 (M+H).

Analysis condition B: residence time = 2.49 min; ESI-MS (-) m/z 923.6 (M-2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5171

Example 5171 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 31.7 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis condition A: residence time = 1.551 min; ESI-MS (+) m/z 944.95 (M+2H).

Analysis conditions B: residence time = 3.165 min; ESI-MS (+) m/z 943.2 (M+2H).

ESI-HRMS (+) m/z: Calculated: 94.

Preparation of Example 5174

Example 5174 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.4 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.58 min; ESI-MS (+) m/z 935.0 (M+2H).

Analysis condition B: residence time = 2.70 min; ESI-MS (-) m/z 933.2 (M- 2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 5175

Example 5175 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM Ammonium acetate; Gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.5 mg and the purity was estimated to be 92% according to LCMS analysis.

Analysis condition A: residence time = 1.408 min; ESI-MS (+) m/z 873.65 (M+2H).

Analysis conditions B: residence time = 2.63 min; ESI-MS (+) m/z 873.7 (M+2H).

Analysis conditions C: residence time = 1.408 min; ESI-MS (+) m/z 873.65 (M+2H).

ESI-HRMS (+) m/z: Calculated: 873.4.

Preparation of Example 5176

Example 5176 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 27.9 mg and its purity was estimated according to LCMS analysis. The value is 99%.

Analysis conditions A: residence time = 1.35 min; ESI-MS (+) m/z 939.5 (M+2H).

Analysis conditions B: residence time = 2.49 min; ESI-MS (+) m/z 939.7 (M+2H).

ESI-HRMS(+) m/z: Calculated value: 938.938 Experimental value: 938.9345.

Preparation of Example 5177

Example 5177 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 27.4 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition A: residence time = 1.393 min; ESI-MS (+) m/z 946.20 (M+2H).

Analysis conditions B: residence time = 2.41 min; ESI-MS (+) m/z 946.2 (M+2H).

ESI-HRMS (+) m/z: Calculated: 945.922.

Preparation of Example 5178

Example 5178 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z 949.7 (M+2H).

Analysis condition B: residence time = 2.59 min; ESI-MS (-) m/z 947.8 (M- 2H).

ESI-HRMS (+) m/z: Calculated: 495.462.

Preparation of Example 5179

Example 5179 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.2 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.39 min; ESI-MS (+) m/z 1847.5 (M+H).

Analysis conditions B: residence time = 2.54 min; ESI-MS (+) m/z 924.6 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5180

Example 5180 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-50% B lasts 30 minutes, then 100% B for 5 minutes; flow rate: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.471 min; ESI-MS (-) m/z 950.95 (M-2H).

Analysis condition B: residence time = 3.060 min; ESI-MS (-) m/z 951.15 (M-2H).

ESI-HRMS (+) m/z: Calculated: 195.

Preparation of Example 5181

Example 5181 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B lasts 30 minutes, then remains at 100% B 5 minutes; flow rate: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 20 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.238 min; ESI-MS (+) m/z 978.95 (M+2H).

Analysis condition B: residence time = 2.659 min; ESI-MS (-) m/z 977.00 (M-2H).

ESI-HRMS (+) m/z: Calculated: 978.4409.

Preparation of Example 5182

Example 5182 was prepared following "General Synthetic Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to use acetic anhydride containing DMF (1.0 M, 2 mL) was used instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 114.3 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.58 min; ESI-MS (+) m/z 912.6 (M+2H).

Analysis conditions B: residence time = 2.65 min; ESI-MS (+) m/z 912.9 (M+2H).

ESI-HRMS (+) m/z: Calculated: 91.

Preparation of Example 5183

Example 5183 was prepared following "General Synthesis Sequence A". The crude material was purified by preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm Granules; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 10-50% B lasts 30 minutes, then remains at 100% B Minute; flow rate: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.9 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis condition A: residence time = 1.61 min; ESI-MS (+) m/z 1772.2 (M+H) analysis condition B: retention time = 2.74 min; ESI-MS (+) m/z 887.0 (M+2H) .

ESI-HRMS (+) m/z: Calculated: 88.

Preparation of Example 5184

Example 5184 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. Combine the fractions containing the desired product and evaporate by centrifugation Dry it. The yield of the product was 19.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.34 min; ESI-MS (-) m/z 915.5 (M-2H).

Analysis conditions B: residence time = 2.48 min; ESI-MS (+) m/z 918.0 (M+2H).

ESI-HRMS (+) m/z: Calculated: 917.4295.

Preparation of Example 5186

Example 5186 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.21 min; ESI-MS (+) m/z 1940.3 (M+H).

Analysis conditions B: residence time = 2.33 min; ESI-MS (+) m/z 970.5 (M+2H).

ESI-HRMS (+) m/z: Calculated: 970.4434.

Preparation of Example 5187

Example 5187 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.2 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.305 min; ESI-MS (-) m/z 955.20 (M-2H).

Analysis conditions B: residence time = 2.639 min; ESI-MS (+) m/z 957.85 (M+2H).

ESI-HRMS (+) m/z: Calculated: 95.

Preparation of Example 5189

Example 5189 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.61 min; ESI-MS (+) m/z 951.4 (M+2H).

Analysis condition B: residence time = 2.91 min; ESI-MS (-) m/z 949.5 (M-2H).

ESI-HRMS (+) m/z: Calculated: 95.

Preparation of Example 5190

Example 5190 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.36 min; ESI-MS (+) m/z 957.0 (M+2H).

Analysis condition B: residence time = 2.52 min; ESI-MS (-) m/z 954.8 (M-2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 5191

Example 5191 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.2 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.599 min; ESI-MS (+) m/z 944.40 (M+2H).

Analysis conditions B: residence time = 1.917 min; ESI-MS (+) m/z 945.20 (M+2H).

ESI-HRMS (+) m/z: Calculated: 944.

Preparation of Example 5192

Example 5192 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 19 x 150 mm, 5 μm particles; mobile phase A: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM Ammonium acetate; Gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.354 min; ESI-MS (+) m/z 938.25 (M+2H).

Analysis conditions B: residence time = 2.876 min; ESI-MS (+) m/z 938.90 (M+2H).

ESI-HRMS (+) m/z: Calculated: 384.446.

Preparation of Example 5196

Example 5196 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.8 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.417 min; ESI-MS (+) m/z 947.15 (M+2H).

Analysis condition B: residence time = 2.469 min; ESI-MS (+) m/z 946.25 (M+2H).

ESI-HRMS (+) m/z: Calculated: 945.922.

Preparation of Example 5198

Example 5198 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.1 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.44 min; ESI-MS (+) m/z 898.7 (M+2H).

Analysis conditions B: residence time = 2.56 min; ESI-MS (+) m/z 899.0 (M+2H).

ESI-HRMS (+) m/z: Calculated: 898.4223.

Preparation of Example 5199

Example 5199 was prepared following "General Synthesis Sequence B". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.4 mg and the purity was estimated to be 96% according to LCMS analysis.

Analytical conditions A: residence time = 1.484 min; ESI-MS (-) m/z 921.40 (M-2H).

Analysis conditions B: residence time = 3.064 min; ESI-MS (+) m/z 923.45 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5200

Example 5200 was prepared following "General Synthetic Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.47 min; ESI-MS (+) m/z 931.3 (M+2H).

Analysis conditions B: residence time = 2.62 min; ESI-MS (+) m/z 931.3 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5201

Example 5201 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19×mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.33 min; ESI-MS (+) m/z 956.0 (M+2H).

Analysis conditions B: residence time = 2.51 min; ESI-MS (+) m/z 955.8 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5202

Example 5202 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.8 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.54 min; ESI-MS (+) m/z 984.3 (M+2H).

Analysis conditions B: residence time = 2.70 min; ESI-MS (+) m/z 984.2 (M+2H).

ESI-HRMS (+) m/z: Calculated: 983.

Preparation of Example 5203

Example 5203 was prepared following "General Synthetic Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.4 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis condition A: residence time = 1.225 min; ESI-MS (-) m/z 956.15 (M-2H).

Analysis condition B: residence time = 2.460 min; ESI-MS (-) m/z 955.15 (M-2H).

ESI-HRMS (+) m/z: Calculated: 95.

Preparation of Example 5204

Example 5204 was prepared following "General Synthetic Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19×mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.39 min; ESI-MS (+) m/z 1848.3 (M+H).

Analysis condition B: residence time = 2.51 min; ESI-MS (-) m/z 922.7 (M-2H).

Preparation of Example 5205

Example 5205 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-50% B for 5 minutes, followed by 100% B for 30 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.36 min; ESI-MS (-) m/z 923.9 (M-2H).

Analysis conditions B: residence time = 2.45 min; ESI-MS (+) m/z 925.1 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5207

Example 5207 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.39 min; ESI-MS (+) m/z 931.6 (M+2H).

Analysis conditions B: residence time = 2.58 min; ESI-MS (+) m/z 931.5 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 5208

Example 5208 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.5 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.35 min; ESI-MS (+) m/z 925.0 (M+2H).

Analysis conditions B: residence time = 2.51 min; ESI-MS (+) m/z 924.7 (M+2H).

ESI-HRMS (+) m/z: Calculated: 923.9327.

Preparation of Example 5209

Example 5209 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 35.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.297 min; ESI-MS (+) m/z 971.45 (M+2H).

Analysis conditions B: residence time = 2.45 min; ESI-MS (+) m/z 971.8 (M+2H).

Analysis conditions C: residence time = 1.297 min; ESI-MS (+) m/z 971.45 (M+2H).

ESI-HRMS (+) m/z: Calculated: 97.

Preparation of Example 5211

Example 5211 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-100% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 25-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.74 min; ESI-MS (+) m/z 957.6 (M+2H).

Analysis conditions B: residence time = 2.98 min; ESI-MS (+) m/z 957.6 (M+2H).

ESI-HRMS (+) m/z: Calculated: 956.9473.

Preparation of Example 5213

Example 5213 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.30 min; ESI-MS (+) m/z 938.2 (M+2H).

Analysis conditions B: residence time = 2.46 min; ESI-MS (+) m/z 938.6 (M+2H).

ESI-HRMS (+) m/z: Calculated: 937.954.

Preparation of Example 5214

Example 5214 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 25-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.39 min; ESI-MS (+) m/z 925.5 (M+2H).

Analysis conditions B: residence time = 2.49 min; ESI-MS (+) m/z 925.5 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5215

Example 5215 was prepared following "General Synthetic Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.44 min; ESI-MS (+) m/z 958.7 (M+2H).

Analysis conditions B: residence time = 2.62 min; ESI-MS (+) m/z 959.2 (M+2H).

ESI-HRMS (+) m/z: Calculated: 95.

Preparation of Example 5216

Example 5216 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.38 min; ESI-MS (+) m/z 938.7 (M+2H).

Analysis conditions B: residence time = 2.49 min; ESI-MS (+) m/z 938.6 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5218

Example 5218 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 20 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.2 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.38 min; ESI-MS (+) m/z 357.2 (M+2H).

Analysis conditions B: residence time = 2.52 min; ESI-MS (+) m/z 956.8 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 5219

Example 5219 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.6 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.52 min; ESI-MS (+) m/z 858.1 (M+2H).

Analysis conditions B: residence time = 2.55 min; ESI-MS (+) m/z 858.7 (M+2H).

ESI-HRMS (+) m/z: Calculated: s.

Preparation of Example 5220

Example 5220 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 40.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analytical condition A: residence time = 1.342 min; ESI-MS (-) m/z 962.60 (M-2H).

Analysis conditions B: residence time = 2.856 min; ESI-MS (+) m/z 965.00 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5221

Example 5221 was prepared following "General Synthetic Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.40 min; ESI-MS (+) m/z 932.1 (M+2H).

Analysis conditions B: residence time = 2.55 min; ESI-MS (+) m/z 932.1 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 5222

Example 5222 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.31 min; ESI-MS (+) m/z 964.1 (M+2H).

Analysis conditions B: residence time = 2.48 min; ESI-MS (+) m/z 963.6 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5223

Example 5223 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.42 min; ESI-MS (+) m/z 956.1 (M+2H).

Analysis conditions B: residence time = 2.59 min; ESI-MS (+) m/z 956.6 (M+2H).

ESI-HRMS (+) m/z: Calculated: 955.

Preparation of Example 5224

Example 5224 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.37 min; ESI-MS (+) m/z 932.1 (M+2H).

Analysis conditions B: residence time = 2.53 min; ESI-MS (+) m/z 931.8 (M+2H).

ESI-HRMS (+) m/z: Calculated: 93.

Preparation of Example 5225

Example 5225 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.6 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.69 min; ESI-MS (+) m/z 944.7 (M+2H).

Analysis conditions B: residence time = 2.90 min; ESI-MS (+) m/z 944.3 (M+2H).

ESI-HRMS (+) m/z: Calculated: 943.9395.

Preparation of Example 5226

Example 5226 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 20 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.375 min; ESI-MS (-) m/z 930.30 (M-2H).

Analysis condition B: residence time = 1.988 min; ESI-MS (+) m/z 932.35 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 5227

Example 5227 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 20 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.429 min; ESI-MS (-) m/z 922.45 (M-2H).

Analysis condition B: residence time = 2.914 min; ESI-MS (+) m/z 924.45 (M+2H).

ESI-HRMS (+) m/z: Calculated: 924.4429.

Preparation of Example 5228

Example 5228 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.58 min; ESI-MS (+) m/z 969.8 (M+2H).

Analysis condition B: residence time = 2.72 min; ESI-MS (-) m/z 267.1 (M-2H).

ESI-HRMS (+) m/z: Calculated: 968.998.

Preparation of Example 5229

Example 5229 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.0 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.41 min; ESI-MS (+) m/z 971.8 (M+2H).

Analysis conditions B: residence time = 2.53 min; ESI-MS (+) m/z 972.1 (M+2H).

ESI-HRMS (+) m/z: Calculated: 97.

Preparation of Example 5230

Example 5230 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.32 min; ESI-MS (+) m/z 950.8 (M+2H).

Analysis conditions B: residence time = 2.40 min; ESI-MS (+) m/z 951.1 (M+2H).

ESI-HRMS (+) m/z: Calculated: 95.

Preparation of Example 5231

Example 5231 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.8 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.31 min; ESI-MS (+) m/z 932.1 (M+2H).

Analysis conditions B: residence time = 2.44 min; ESI-MS (+) m/z 932.4 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 5232

Example 5232 was prepared following "General Synthetic Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.34 min; ESI-MS (+) m/z 937.9 (M+2H).

Analysis conditions B: residence time = 2.47 min; ESI-MS (-) m/z 936.2 (M-2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5233

Example 5233 was prepared following "General Synthetic Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 methanol: water + 0.1% trifluoroacetic acid; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles: mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.29 min; ESI-MS (+) m/z 925.8 (M+2H).

Analysis conditions B: residence time = 2.40 min; ESI-MS (-) m/z 924.1 (M-2H).

Preparation of Example 5234

Example 5234 was prepared following "General Synthetic Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.61 min; ESI-MS (+) m/z 935.6 (M+2H).

Analysis condition B: residence time = 2.71 min; ESI-MS (-) m/z 933.7 (M-2H).

ESI-HRMS (+) m/z: Calculated: 935.4589.

Preparation of Example 5236

Example 5236 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 25-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.21 min; ESI-MS (+) m/z 972.9 (M+2H).

Analysis conditions B: residence time = 2.31 min; ESI-MS (+) m/z 972.0 (M+2H).

ESI-HRMS (+) m/z: Calculated: 97.

Preparation of Example 5237

Example 5237 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 150 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.43 min; ESI-MS (+) m/z 931.0 (M+2H).

Analysis conditions B: residence time = 2.58 min; ESI-MS (+) m/z 931.0 (M+2H).

ESI-HRMS (+) m/z: Calculated: 930.4485.

Preparation of Example 5238

Example 5238 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B : 95:5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 25-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.7 mg and the purity was estimated to be 93% according to LCMS analysis.

Analysis conditions A: residence time = 1.29 min; ESI-MS (+) m/z 964.1 (M+2H).

Analysis conditions B: residence time = 2.46 min; ESI-MS (+) m/z 964.2 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5240

Example 5240 was prepared following "General Synthetic Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.4 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.30 min; ESI-MS (+) m/z 946.0 (M+2H).

Analysis conditions B: residence time = 2.44 min; ESI-MS (+) m/z 946.2 (M+2H).

ESI-HRMS (+) m/z: Calculated: 945.43.

Preparation of Example 5241

Example 5241 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.3 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.51 min; ESI-MS (+) m/z 978.5 (M+2H).

Analysis conditions B: residence time = 2.77 min; ESI-MS (-) m/z 975.3 (M-2H).

ESI-HRMS (+) m/z: Calculated: 976.9774.

Preparation of Example 5242

Example 5242 was prepared following "General Synthetic Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.34 min; ESI-MS (+) m/z 1849.3 (M+H).

Analysis condition B: residence time = 2.47 min; ESI-MS (-) m/z 923.1 (M-2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5243

Example 5243 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 Methanol: water + 0.1% trifluoroacetic acid; gradient: 40-100% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.22 min; ESI-MS (+) m/z 946.0 (M+2H).

Analysis condition B: residence time = 2.31 min; ESI-MS (-) m/z 944.1 (M-2H).

ESI-HRMS (+) m/z: Calculated: 945.43.

Preparation of Example 5245

Example 5245 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.6 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.576 min; ESI-MS (+) m/z 946.60 (M+2H).

Analysis conditions B: residence time = 2.171 min; ESI-MS (+) m/z 945.55 (M+2H).

Analysis conditions C: residence time = 1.576 min; ESI-MS (+) m/z 946.60 (M+2H).

ESI-HRMS (+) m/z: Calculated: 945.43.

Preparation of Example 5246

Example 5246 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.42 min; ESI-MS (-) m/z 923.0 (M-2H).

Analysis conditions B: residence time = 2.54 min; ESI-MS (+) m/z 925.1 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5247

Example 5247 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.0 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.49 min; ESI-MS (+) m/z 924.0 (M+2H).

Analysis conditions B: residence time = 2.64 min; ESI-MS (-) m/z 922.1 (M-2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5248

Example 5248 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 60-100% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.7 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.65 min; ESI-MS (+) m/z 1847.4 (M+H).

Analysis conditions B: residence time = 2.93 min; ESI-MS (+) m/z 924.5 (M+2H).

Preparation of Example 5249

Example 5249 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 37.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.06 min; ESI-MS (+) m/z 943.5 (M+2H).

Analysis conditions B: residence time = 2.16 min; ESI-MS (-) m/z 941.4 (M-2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5251

Example 5251 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.49 min; ESI-MS (+) m/z 931.5 (M+2H).

Analysis conditions B: residence time = 2.61 min; ESI-MS (-) m/z 929.2 (M-2H).

ESI-HRMS (+) m/z: Calculated: 303.

Preparation of Example 5252

Example 5252 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.30 min; ESI-MS (+) m/z 932.3 (M+2H).

Analysis condition B: residence time = 2.45 min; ESI-MS (-) m/z 930.6 (M-2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5253

Example 5253 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 20 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 20 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.378 min; ESI-MS (+) m/z 931.9 (M+2H).

Analysis conditions B: residence time = 2.862 min; ESI-MS (-) m/z 929.9 (M-2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 5254

Example 5254 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.655 min; ESI-MS (+) m/z 938.60 (M+2H).

Analysis condition B: residence time = 2.40 min; ESI-MS (+) m/z 939.2 (M+2H).

Analysis conditions C: residence time = 1.655 min; ESI-MS (+) m/z 938.60 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5255

Example 5255 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 25- 75% B lasted 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.32 min; ESI-MS (+) m/z 971.2 (M+2H).

Analysis conditions B: residence time = 2.44 min; ESI-MS (+) m/z 971.6 (M+2H).

ESI-HRMS (+) m/z: Calculated: 97.

Preparation of Example 5256

Example 5256 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: tube Column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B duration 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.54 min; ESI-MS (+) m/z 993.1 (M+2H).

Analysis conditions B: residence time = 2.73 min; ESI-MS (+) m/z 992.8 (M+2H).

ESI-HRMS (+) m/z: Calculated: 992.4897.

Preparation of Example 5257

Example 5257 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. Combine the fractions containing the desired product and evaporate by centrifugation Dry it. The yield of the product was 17.2 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.34 min; ESI-MS (+) m/z 1876.4 (M+2H).

Analysis condition B: residence time = 2.45 min; ESI-MS (-) m/z 936.3 (M-2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 5259

Example 5259 was prepared following "General Synthesis Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Prelude Method A" were modified to replace pure acetic anhydride with DMF (1.0 M, 2 mL) containing acetic anhydride. mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. Combine the fractions containing the desired product and steam by centrifugation Hair is dried. The yield of the product was 31.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z 951.3 (M+2H).

Analysis condition B: residence time = 2.69 min; ESI-MS (-) m/z 950.0 (M-2H).

ESI-HRMS (+) m/z: Calculated: 195.

Preparation of Example 5260

Example 5260 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.9 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.61 min; ESI-MS (+) m/z 928.2 (M+2H).

Analysis conditions B: residence time = 2.72 min; ESI-MS (+) m/z 928.6 (M+2H).

Analysis conditions C: residence time = 1.432 min; ESI-MS (+) m/z 946.70 (M+2H).

ESI-HRMS (+) m/z: Calculated: 940.

Preparation of Example 5261

Example 5261 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.6 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.432 min; ESI-MS (+) m/z 946.70 (M+2H).

Analysis conditions B: residence time = 2.462 min; ESI-MS (-) m/z 944.80 (M-2H).

ESI-HRMS (+) m/z: Calculated: 940.

Preparation of Example 5262

Example 5262 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.62 min; ESI-MS (+) m/z 908.8 (M+2H).

Analysis condition B: residence time = 2.75 min; ESI-MS (-) m/z 907.1 (M-2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5263

Example 5263 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.43 min; ESI-MS (+) m/z 920.9 (M+2H).

Analysis conditions B: residence time = 2.59 min; ESI-MS (-) m/z 919.2 (M-2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 5264

Example 5264 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.82 min; ESI-MS (=) m/z 964.9 (M+2H).

Analysis condition B: residence time = 2.94 min; ESI-MS (+) m/z 964.6 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 5265

Example 5265 was prepared following "General Synthetic Sequence A", wherein the "Single Coupling Procedure" and "Double Coupling Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.631 min; ESI-MS (-) m/z 920.30 (M-2H).

Analysis condition B: residence time = 3.126 min; ESI-MS (+) m/z 922.35 (M+2H).

Preparation of Example 6118

Example 6118 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.52 min; ESI-MS (+) m/z 965.3 (M+2H).

Analysis condition B: residence time = 2.16 min; ESI-MS (+) m/z 965.65 (M+2H).

ESI-HRMS (+) m/z: Calculated: 965.4.

Preparation of Example 6119

Example 6119 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product is 2.9 mg, and an estimated purity of 96% based on LCMS analysis.

Analysis conditions A: residence time = 1.27 min; ESI-MS (+) m/z 938.75 (M+2H).

Analysis condition B: residence time = 2.79 min; ESI-MS (+) m/z 939 (M+2H).

ESI-HRMS (+) m/z: Calculated: 384.

Preparation of Example 6128

Example 6128 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.4 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition A: residence time = 1.43 min; ESI-MS (+) m/z 956.0 (M+2H).

Analysis conditions B: residence time = 2.89 min; ESI-MS (+) m/z 956.0 (M+2H).

ESI-HRMS (+) m/z: Calculated: 955.4.

Preparation of Example 6134

Example 6134 was prepared following "General Synthetic Sequence A", wherein the "Single Coupling Procedure" and "Double Coupling Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B : 95:5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.4 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis condition A: residence time = 1.22 min; ESI-MS (-) m/z 984.4 (M+2H).

Analysis conditions B: residence time = 2.30 min; ESI-MS (-) m/z 984.6 (M+2H).

ESI-HRMS (+) m/z: Calculated: 985.4487.

Preparation of Example 6135

Example 6135 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 0.1% trifluoroacetic acid; mobile phase B : 95:5 methanol: water + 0.1% trifluoroacetic acid; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.9 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.18 min; ESI-MS (+) m/z 954.2 (M+2H).

Analysis conditions B: residence time = 2.27 min; ESI-MS (+) m/z 953.5 (M+2H).

ESI-HRMS (+) m/z: Calculated: 195.

Preparation of Example 6136

Example 6136 was prepared following "General Synthetic Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.30 min; ESI-MS (+) m/z 952.9 (M+2H).

Analysis condition B: residence time = 1.90 min; ESI-MS (+) m/z 952.7 (M+2H).

ESI-HRMS (+) m/z: Calculated: 195.

Preparation of Example 6137

Example 6137 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.0 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.31 min; ESI-MS (+) m/z 945.95 (M+2H).

ESI-HRMS (+) m/z: Calculated: 945.

Preparation of Example 6140

Example 6140 was prepared following "General Synthesis Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.36 min; ESI-MS (+) m/z 896.4 (M+2H).

Analysis conditions B: residence time = 2.56 min; ESI-MS (+) m/z 896.4 (M+2H).

ESI-HRMS (+) m/z: Calculated: 895.4402.

Preparation of Example 6141

Example 6141 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.44 min; ESI-MS (+) m/z 978.5 (M+2H).

Analysis conditions B: residence time = 2.94 min; ESI-MS (+) m/z 978.4 (M+2H).

ESI-HRMS (+) m/z: Calculated: 97.

Preparation of Example 6146

Example 6146 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.52 min; ESI-MS (+) m/z 952.4 (M+2H).

Analysis conditions B: residence time = 3.07 min; ESI-MS (+) m/z 952.4 (M+2H).

ESI-HRMS (+) m/z: Calculated: 195.

Preparation of Example 6156

Example 6156 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.9 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.46 min; ESI-MS (+) m/z 992.0 (M+2H).

Analysis conditions B: residence time = 2.98 min; ESI-MS (+) m/z 992.0 (M+2H).

ESI-HRMS (+) m/z: Calculated: 99.

Preparation of Example 6158

Example 6158 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95 : 5 acetonitrile: water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. Combined It contains the soluble fraction of the desired product and is dried by centrifugal evaporation. The yield of the product was 17.3 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.30 min; ESI-MS (+) m/z 957.1 (M+2H).

Analysis conditions B: residence time = 2.43 min; ESI-MS (+) m/z 956.9 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 6162

Example 6162 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.9 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.21 min; ESI-MS (+) m/z 999.7 (M+2H).

Analysis conditions B: residence time = 2.68 min; ESI-MS (+) m/z 999.6 (M+2H).

ESI-HRMS (+) m/z: Calculated: 999.4.

Preparation of Example 6168

Example 6168 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 150 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.51 min; ESI-MS (+) m/z 920.65 (M+2H).

Analysis condition B: residence time = 2.78 min; ESI-MS (+) m/z 920.65 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 6170

Example 6170 was prepared following "General Synthesis Sequence A", wherein the "Single Coupling Procedure" and "Double Coupling Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. Combine the fractions containing the desired product and steam by centrifugation Hair is dried. The yield of the product was 7.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.55 min; ESI-MS (+) m/z 924.3 (M+2H).

Analysis conditions B: residence time = 2.87 min; ESI-MS (+) m/z 924.3 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 6172

Example 6172 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water +10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.21 min; ESI-MS (+) m/z 960.7 (M+2H).

Analysis condition B: residence time = 1.79 min; ESI-MS (+) m/z 960.9 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 6176

Example 6176 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. Combine the fractions containing the desired product and evaporate by centrifugation Dry it. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.47 min; ESI-MS (+) m/z 971.0 (M+2H).

Analysis conditions B: residence time = 2.91 min; ESI-MS (+) m/z 970.9 (M+2H).

Preparation of Example 6180

Example 6180 was prepared following "General Synthetic Sequence A", wherein the "Single Coupling Procedure" and "Double Coupling Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B duration 30 Minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.50 min; ESI-MS (-) m/z 907.7 (M-2H).

Analysis condition B: residence time = 2.71 min; ESI-MS (+) m/z 910.3 (M+2H).

ESI-HRMS (+) m/z: Calculated: 908.9582.

Preparation of Example 6183

Example 6183 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 0.1% trifluoroacetic acid; mobile phase B : 95:5 methanol: water + 0.1% trifluoroacetic acid; gradient: 40-80% B for 30 minutes, followed by 100% B for 50 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material is further purified via preparative LC/MS using the following conditions Tube: column: XBridge C18, 19 × 200mm, 5μm particles; mobile phase A: 5:95 methanol: water + 10mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10mM ammonium acetate; gradient: 30-70 % B lasted 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.3 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.24 min; ESI-MS (+) m/z 957.8 (M+2H).

Analysis conditions B: residence time = 2.36 min; ESI-MS (+) m/z 957.6 (M+2H).

Preparation of Example 6184

Example 6184 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. Combine the fractions containing the desired product and evaporate by centrifugation Dry it. The yield of the product was 8.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.25 min; ESI-MS (-) m/z 958.8 (M-2H).

Analysis conditions B: residence time = 1.62 min; ESI-MS (+) m/z 960.5 (M+2H).

ESI-HRMS (+) m/z: Calculated: 959.4.

Preparation of Example 6185

Example 6185 was prepared following "General Synthetic Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. Substance through the use of the following conditions Prepared LC/MS for further purification: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate Gradient: 10-50% B for 30 minutes followed by 100 minutes at 100% B; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.0 mg and the purity was estimated to be 92% according to LCMS analysis.

Analysis conditions A: residence time = 1.38 min; ESI-MS (-) m/z 976.9 (M-2H).

Analysis conditions B: residence time = 3.09 min; ESI-MS (+) m/z 977.6 (M+2H).

ESI-HRMS (+) m/z: Calculated: 977.4.

Preparation of Example 6189

Example 6189 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 37.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.18 min; ESI-MS (+) m/z 960.5 (M+2H).

Analysis conditions B: residence time = 2.33 min; ESI-MS (+) m/z 960.7 (M+2H).

ESI-HRMS (+) m/z: Calculated: 95.

Preparation of Example 6201

Example 6201 was prepared following "General Synthetic Sequence A". The crude material was purified by preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm Granules; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B lasts 30 minutes, then remains at 100% B Minute; flow rate: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.7 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.49 min; ESI-MS (+) m/z 951.6 (M+2H).

Analysis conditions B: residence time = 3.06 min; ESI-MS (+) m/z 951.6 (M+2H).

Preparation of Example 6202

Example 6202 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. Combine the fractions containing the desired product and evaporate by centrifugation Dry it. The yield of the product was 17.1 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.27 min; ESI-MS (+) m/z 972.3 (M+2H).

Analysis conditions B: residence time = 2.44 min; ESI-MS (+) m/z 972.1 (M+2H).

ESI-HRMS (+) m/z: Calculated: 97.

Preparation of Example 6203

Example 6203 was prepared following "General Synthetic Sequence A", wherein the "Single Coupling Procedure" and "Double Coupling Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm Granules; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 5-45% B for 20 minutes, then at 100% B Minute; flow rate: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.39 min; ESI-MS (-) m/z 929.0 (M-2H).

Analysis conditions B: residence time = 2.51 min; ESI-MS (+) m/z 931.1 (M+2H).

ESI-HRMS (+) m/z: Calcd.: 992.

Preparation of Example 6204

Example 6204 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then 5 minutes at 100% B Clock; flow: 20 mL / min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.43 min; ESI-MS (+) m/z 991.5 (M+2H).

Analysis conditions B: residence time = 2.98 min; ESI-MS (+) m/z 991.5 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 6207

Example 6207 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.50 min; ESI-MS (-) m/z 907.7 (M-2H).

Analysis condition B: residence time = 2.71 min; ESI-MS (+) m/z 910.3 (M+2H).

ESI-HRMS (+) m/z: Calculated: 908.9582.

Preparation of Example 6209

Example 6209 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.28 min; ESI-MS (+) m/z 992.0 (M+2H).

Analysis conditions B: residence time = 2.69 min; ESI-MS (+) m/z 993.0 (M+2H).

ESI-HRMS(+) m/z: Calculated value: 992.4747 Experimental value: 992.4713

Preparation of Example 6213

Example 6213 was prepared following "General Synthesis Sequence A", wherein "Single Coupling Procedure" and "Double Coupling Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.2 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.32 min; ESI-MS (-) m/z 959.5 (M-2H).

Analysis conditions B: residence time = 2.51 min; ESI-MS (-) m/z 959.8 (M-2H).

ESI-HRMS (+) m/z: Calculated: s.

Preparation of Example 6216

Example 6216 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.9 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.45 min; ESI-MS (+) m/z 945.4 (M+2H).

Analysis conditions B: residence time = 2.95 min; ESI-MS (+) m/z 945.6 (M+2H).

ESI-HRMS (+) m/z: Calculated: 945.

Preparation of Example 6224

Example 6224 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 0.1% trifluoroacetic acid; mobile phase B : 95:5 methanol: water + 0.1% trifluoroacetic acid; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.0 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.29 min; ESI-MS (+) m/z 1013.45 (M+2H).

Analysis condition B: residence time = 0.42 min; ESI-MS (+) m/z 1013.7 (M+2H).

Preparation of Example 6228

Example 6228 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 20 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.0 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.34 min; ESI-MS (+) m/z 992.6 (M+2H).

Analysis condition B: residence time = 1.85 min; ESI-MS (+) m/z 992.7 (M+2H).

ESI-HRMS (+) m/z: Calculated: 992.4622.

Preparation of Example 6236

Example 6236 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.5 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition A: residence time = 1.42 min; ESI-MS (+) m/z 962.4 (M+2H).

Analysis conditions B: residence time = 2.92 min; ESI-MS (+) m/z 964.5 (M+2H).

ESI-HRMS (+) m/z: Calculated: 963.

Preparation of Example 6244

Example 6244 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product is 10.0 mg, and its purity estimate was 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.35 min; ESI-MS (+) m/z 957.6 (M+2H).

Analysis conditions B: residence time = 2.82 min; ESI-MS (+) m/z 957.8 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 6247

Example 6247 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35- 75% B lasted 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.34 min; ESI-MS (+) m/z 943.0 (M+2H).

Analysis conditions B: residence time = 1.89 min; ESI-MS (+) m/z 942.9 (M+2H).

ESI-HRMS (+) m/z: Calculated: 149.

Preparation of Example 6249

Example 6249 was prepared following "General Synthetic Sequence A", wherein the "Single Coupling Procedure" and "Double Coupling Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B duration 30 Minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.36 min; ESI-MS (+) m/z 943.4 (M+2H).

Analysis conditions B: residence time = 2.82 min; ESI-MS (+) m/z 943.4 (M+2H).

ESI-HRMS (+) m/z: Calculated: 94.

Preparation of Example 6250

Example 6250 was prepared following "General Synthesis Sequence A". Crude material by using the following Preparative LC/MS purification: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM acetic acid Ammonium; Gradient: 30-70% B for 30 minutes followed by 100 minutes at 100% B; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.24 min; ESI-MS (+) m/z 946.1 (M+2H).

Analysis condition B: residence time = 2.61 min; ESI-MS (+) m/z 946.1 (M+2H).

ESI-HRMS (+) m/z: Calculated: 495.

Preparation of Example 6258

Example 6258 is prepared following "General Synthetic Sequence A", in which "Symphony Method The "single coupling procedure" and "double coupling procedure" of A" were modified to replace pure acetic anhydride (2.0 mL) with DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.2 mg and the purity was estimated to be 93% according to LCMS analysis.

Analysis conditions A: residence time = 1.44 min; ESI-MS (+) m/z 922.8 (M+2H).

Analysis conditions B: residence time = 2.93 min; ESI-MS (+) m/z 923.1 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 6262

Example 6262 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.9 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.47 min; ESI-MS (+) m/z 928.7 (M+2H).

Analysis conditions B: residence time = 2.74 min; ESI-MS (+) m/z 928.7 (M+2H).

ESI-HRMS (+) m/z: Calculated: s.

Preparation of Example 6266

Example 6266 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.5 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.49 min; ESI-MS (+) m/z 952.9 (M+2H).

Analysis conditions B: residence time = 3.09 min; ESI-MS (+) m/z 952.7 (M+2H).

ESI-HRMS (+) m/z: Calculated: 195.

Preparation of Example 6267

Example 6267 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 20 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.26 min; ESI-MS (+) m/z 992.7 (M+2H).

Analysis conditions B: residence time = 2.75 min; ESI-MS (+) m/z 992.7 (M+2H).

Preparation of Example 6269

Example 6269 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.19 min; ESI-MS (+) m/z 972.9 (M+2H).

Analysis condition B: residence time = 2.32 min; ESI-MS (+) m/z 972.7 (M+2H).

ESI-HRMS (+) m/z: Calculated: 97.

Preparation of Example 6272

Example 6272 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.7 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.47 min; ESI-MS (+) m/z 896.1 (M+2H).

Analysis condition B: residence time = 2.63 min; ESI-MS (+) m/z 896.1 (M+2H).

ESI-HRMS (+) m/z: Calculated: 895.4765.

Preparation of Example 6274

Example 6274 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.29 min; ESI-MS (+) m/z 925.5 (M+2H).

Analysis condition B: residence time = 2.68 min; ESI-MS (+) m/z 925.9 (M+2H).

ESI-HRMS (+) m/z: Calculated: 925.4.

Preparation of Example 6276

Example 6276 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.29 min; ESI-MS (-) m/z 970.9 (M- 2H).

Analysis conditions B: residence time = 1.55 min; ESI-MS (+) m/z 972.5 (M+2H).

ESI-HRMS (+) m/z: Calculated: 971.4456.

Preparation of Example 6283

Example 6283 was prepared following "General Synthetic Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.3 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition B: residence time = 3.22 min; ESI-MS (+) m/z 936.1 (M+2H).

Analysis conditions C: residence time = 1.63 min; ESI-MS (+) m/z 935.9 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 6284

Example 6284 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-100% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product is 18.3 mg, and an estimated purity of 98% based on LCMS analysis.

Analysis conditions A: residence time = 1.55 min; ESI-MS (+) m/z 958.3 (M+2H).

Analysis conditions B: residence time = 2.22 min; ESI-MS (+) m/z 958.2 (M+2H).

ESI-HRMS (+) m/z: Calculated: 957.982.

Preparation of Example 6288

Example 6288 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.24 min; ESI-MS (+) m/z 985.2 (M+2H).

Analysis conditions B: residence time = 1.80 min; ESI-MS (+) m/z 985.1 (M+2H).

ESI-HRMS (+) m/z: Calculated: 98.

Preparation of Example 6289

Example 6289 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product is 12.1 mg, and its purity estimate was 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.24 min; ESI-MS (+) m/z 953.9 (M+2H).

Analysis conditions B: residence time = 2.57 min; ESI-MS (+) m/z 953.9 (M+2H).

ESI-HRMS (+) m/z: Calculated: 95.

Preparation of Example 6293

Example 6293 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.30 min; ESI-MS (+) m/z 971.0 (M+2H).

Analysis conditions B: residence time = 2.74 min; ESI-MS (+) m/z 971.0 (M+2H).

ESI-HRMS (+) m/z: Calculated: 970.4672.

Preparation of Example 6296

Example 6296 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.0 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.46 min; ESI-MS (+) m/z 958.5 (M+2H).

Analysis condition B: residence time = 3.01 min; ESI-MS (+) m/z 958.5 (M+2H).

ESI-HRMS (+) m/z: Calculated: 958.

Preparation of Example 6297

Example 6297 was prepared following "General Synthesis Sequence A". The crude material was purified by preparative LC/MS using the following conditions: column: XBridge C18, 19×mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.32 min; ESI-MS (+) m/z 978.9 (M+2H).

Analysis conditions B: residence time = 2.77 min; ESI-MS (+) m/z 978.9 (M+2H).

ESI-HRMS (+) m/z: Calculated: 978.

Preparation of Example 6301

Example 6301 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product is 38.9 mg, and an estimated purity of 100% based on LCMS analysis.

Analysis conditions A: residence time = 1.34 min; ESI-MS (+) m/z 938.9 (M+2H).

Analysis conditions B: residence time = 2.84 min; ESI-MS (+) m/z 938.8 (M+2H).

ESI-HRMS (+) m/z: Calculated: 384.446.

Preparation of Example 6303

Example 6303 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10- 50% B lasted 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.34 min; ESI-MS (+) m/z 930.9 (M+2H).

Analysis conditions B: residence time = 2.85 min; ESI-MS (+) m/z 930.9 (M+2H).

ESI-HRMS (+) m/z: Calculated: 930.4.

Preparation of Example 6309

Example 6309 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: tube Column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 5-45% B duration 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.39 min; ESI-MS (+) m/z 924.1 (M+2H).

Analysis conditions B: residence time = 2.91 min; ESI-MS (+) m/z 924.1 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 6310

Example 6310 was prepared following "General Synthetic Sequence A", wherein the "Single Coupling Procedure" and "Double Coupling Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. Substance through the use of the following conditions Prepared LC/MS for further purification: column: XBridge C18, 19 × 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate Gradient: 45-85% B for 20 minutes followed by 100 minutes at 100% B; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.7 mg and the purity was estimated to be 94% according to LCMS analysis.

Analysis conditions A: residence time = 1.54 min; ESI-MS (+) m/z 916.7 (M+2H).

Analysis condition B: residence time = 2.82 min; ESI-MS (+) m/z 916.5 (M+2H).

ESI-HRMS (+) m/z: Calculated: 915.

Preparation of Example 6324

Example 6234 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol :water+ 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, followed by 100 minutes at 100% B; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.24 min; ESI-MS (+) m/z 1000.0 (M+2H).

Analysis conditions B: residence time = 2.69 min; ESI-MS (+) m/z 999.2 (M+2H).

Preparation of Example 6131

Example 6131 was prepared following "General Synthesis Sequence A". The crude material was purified by preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm Granules; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B lasts 30 minutes, then remains at 100% B Minute; flow rate: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.32 min; ESI-MS (+) m/z 971.2 (M+2H).

Analysis conditions B: residence time = 2.89 min; ESI-MS (+) m/z 971.2 (M+2H).

ESI-HRMS (+) m/z: Calculated: 978.959.

Preparation of Example 6334

Prepare Example 6334 following "General Synthetic Sequence A", in which "Symphony Method The "single coupling procedure" and "double coupling procedure" of A" were modified to replace pure acetic anhydride (2.0 mL) with DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.8 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.32 min; ESI-MS (+) m/z 910.2 (M+2H).

Analysis conditions B: residence time = 2.77 min; ESI-MS (+) m/z 910.3 (M+2H).

ESI-HRMS (+) m/z: Calculated: 909.9296.

Preparation of Example 6335

Example 6335 was prepared following "General Synthesis Sequence A". Crude material by using the following Preparative LC/MS purification: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM acetic acid Ammonium; Gradient: 25-70% B for 30 minutes followed by 100 minutes at 100% B; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.19 min; ESI-MS (+) m/z 979.4 (M+2H).

Analysis conditions B: residence time = 2.50 min; ESI-MS (+) m/z 979.4 (M+2H).

Preparation of Example 6338

Example 6338 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.33 min; ESI-MS (+) m/z 992.5 (M+2H).

Analysis conditions B: residence time = 2.78 min; ESI-MS (+) m/z 992.5 (M+2H).

ESI-HRMS(+) m/z: Calculated value: 991.9883 Experimental value: 991.9861

Preparation of Example 6344

Example 6344 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.47 min; ESI-MS (+) m/z 952.7 (M+2H).

Analysis conditions B: residence time = 2.37 min; ESI-MS (+) m/z 952.6 (M+2H).

ESI-HRMS (+) m/z: Calculated: 195.

Preparation of Example 6347

Example 6347 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.6 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.29 min; ESI-MS (+) m/z 950.4 (M+2H).

Analysis conditions B: residence time = 2.66 min; ESI-MS (+) m/z 950.4 (M+2H).

ESI-HRMS (+) m/z: Calculated: 149.

Preparation of Example 6350

Example 6350 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.45 min; ESI-MS (+) m/z 965.4 (M+2H).

Analysis conditions B: residence time = 2.95 min; ESI-MS (+) m/z 965.4 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 6361

Example 6361 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. Combined It contains the soluble fraction of the desired product and is dried by centrifugal evaporation. The yield of the product was 13.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.33 min; ESI-MS (+) m/z 985.9 (M+2H).

Analysis conditions B: residence time = 2.78 min; ESI-MS (+) m/z 985.9 (M+2H).

ESI-HRMS (+) m/z: Calculated: 985.4

Preparation of Example 6363

Example 6363 was prepared following "General Synthetic Sequence A", wherein the "Single Coupling Procedure" and "Double Coupling Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. Substance preparation by using the following conditions Further purification by LC/MS: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; Gradient: 35-75% B for 30 minutes followed by 100 minutes at 100% B; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.3 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.36 min; ESI-MS (+) m/z 916.9 (M+2H).

Analysis conditions B: residence time = 2.88 min; ESI-MS (+) m/z 916.9 (M+2H).

ESI-HRMS (+) m/z: Calculated: 916.4454.

Preparation of Example 6365

Example 6365 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol :water+ 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, followed by 100 minutes at 100% B; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 10-50% B for 20 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.2 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.20 min; ESI-MS (-) m/z 963.8 (M-2H).

Analysis conditions B: residence time = 2.32 min; ESI-MS (+) m/z 966.6 (M+2H).

ESI-HRMS (+) m/z: Calculated: 965.9354.

Preparation of Example 6369

Example 6369 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-80% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.34 min; ESI-MS (+) m/z 963.4 (M+2H).

Analysis conditions B: residence time = 2.79 min; ESI-MS (+) m/z 963.4 (M+2H).

ESI-HRMS (+) m/z: Calculated: 962.

Preparation of Example 6370

Example 6370 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 20 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.35 min; ESI-MS (+) m/z 984.9 (M+2H).

Analysis conditions B: residence time = 2.85 min; ESI-MS (+) m/z 984.0 (M+2H).

Preparation of Example 6372

Example 6372 was prepared following "General Synthetic Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 20 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.8 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.51 min; ESI-MS (+) m/z 923.8 (M+2H).

Analysis conditions B: residence time = 2.77 min; ESI-MS (+) m/z 923.7 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 6380

Example 6380 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. Combined It contains the soluble fraction of the desired product and is dried by centrifugal evaporation. The yield of the product was 17.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.33 min; ESI-MS (+) m/z 957.3 (M+2H).

Analysis conditions B: residence time = 2.86 min; ESI-MS (+) m/z 957.3 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 6383

Example 6383 was prepared following "General Synthetic Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.34 min; ESI-MS (+) m/z 903.2 (M+2H).

Analysis conditions B: residence time = 2.55 min; ESI-MS (+) m/z 903.4 (M+2H).

ESI-HRMS (+) m/z: Calculated: 901.

Preparation of Example 6384

Example 6384 was prepared following "General Synthesis Sequence A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 27.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.23 min; ESI-MS (+) m/z 954.1 (M+2H).

Analysis conditions B: residence time = 1.48 min; ESI-MS (+) m/z 954.0 (M+2H).

ESI-HRMS (+) m/z: Calculated: 195.456.

Preparation of Example 6389

Example 6389 was prepared following "General Synthesis Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then 5 minutes at 100% B Clock; flow: 20 mL / min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.8 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.49 min; ESI-MS (+) m/z 936.9 (M+2H).

Analysis condition B: residence time = 2.84 min; ESI-MS (-) m/z 934.9 (M-2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 6390

Example 6390 was prepared following "General Synthesis Sequence A", wherein "Single Coupling Procedure" and "Double Coupling Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B duration 30 Minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z 969.0 (M+2H).

Analysis conditions B: residence time = 2.84 min; ESI-MS (+) m/z 969.2 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 6391

Example 6391 is prepared following "General Synthetic Sequence A", in which "Symphony Method The "single coupling procedure" and "double coupling procedure" of A" were modified to replace pure acetic anhydride (2.0 mL) with DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.30 min; ESI-MS (+) m/z 1005.4 (M+2H).

Analysis condition B: residence time = 2.54 min; ESI-MS (-) m/z 1003.5 (M-2H).

ESI-HRMS (+) m/z: Calculated: 1005.4882.

Preparation of Example 6392

Example 6392 was prepared following "General Synthetic Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.481 min; ESI-MS (+) m/z 969.25 (M+2H).

Analysis condition B: residence time = 3.004 min; ESI-MS (+) m/z 969.80 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 6393

Example 6393 was prepared following "General Synthetic Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.270 min; ESI-MS (+) m/z 953.40 (M+2H).

Analysis condition B: residence time = 2.685 min; ESI-MS (-) m/z 951.55 (M-2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 6394

Example 6394 was prepared following "General Synthetic Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 33.3 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.227 min; ESI-MS (-) m/z 976.85 (M-2H).

Analysis condition B: residence time = 0.366 min; ESI-MS (-) m/z 976.75 (M-2H).

ESI-HRMS (+) m/z: Calculated: 97.

Preparation of Example 6395

Example 6395 was prepared following "General Synthesis Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.0 mg and the purity was estimated to be 93% according to LCMS analysis.

Analysis condition A: residence time = 1.67 min; ESI-MS (+) m/z 978.3 (M+2H).

Analysis conditions B: residence time = 3.09 min; ESI-MS (+) m/z 978.2 (M+2H).

ESI-HRMS (+) m/z: Calculated: 977.4.

Preparation of Example 6396

Example 6936 was prepared following "General Synthetic Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.3 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis condition A: residence time = 1.54 min; ESI-MS (+) m/z 960.9 (M+2H).

Analysis conditions B: residence time = 2.96 min; ESI-MS (+) m/z 961.7 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 6397

Example 6397 was prepared following "General Synthetic Sequence A", wherein the "Single Coupling Procedure" and "Double Coupling Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.47 min; ESI-MS (+) m/z 972.2 (M+2H).

Analysis conditions B: residence time = 2.80 min; ESI-MS (+) m/z 972.2 (M+2H).

ESI-HRMS (+) m/z: Calculated: 97.

Preparation of Example 6398

Example 6398 was prepared following "General Synthetic Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile :water +10 mM ammonium acetate; Gradient: 0-40% B for 30 minutes followed by 100 minutes at 100% B; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.2 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.16 min; ESI-MS (-) m/z 943.4 (M-2H).

Analysis condition B: residence time = 2.43 min; ESI-MS (+) m/z 945.6 (M+2H).

ESI-HRMS (+) m/z: Calculated: s.

Preparation of Example 6399

Example 6399 was prepared following "General Synthesis Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM acetic acid Ammonium; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.5 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.51 min; ESI-MS (+) m/z 961.3 (M+2H).

Analysis conditions B: residence time = 2.92 min; ESI-MS (+) m/z 961.6 (M+2H).

ESI-HRMS(+) m/z: Calculated: 960.9769 Experimental value: 960.9745

Preparation of Example 6400

Example 6400 was prepared following "General Synthetic Sequence A", wherein the "Single Coupling Procedure" and "Double Coupling Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM acetic acid Ammonium; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.34 min; ESI-MS (+) m/z 931.6 (M+2H).

Analysis conditions B: residence time = 2.72 min; ESI-MS (-) m/z 928.9 (M-2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 6401

Example 6401 was prepared following "General Synthetic Sequence A", in which "Single Coupling Procedure" and "Double Coupling Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM acetic acid Ammonium; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.26 min; ESI-MS (-) m/z 993.9 (M-2H).

Analysis conditions B: residence time = 2.51 min; ESI-MS (+) m/z 996.5 (M+2H).

Preparation of Example 6402

Example 6402 was prepared following "General Synthetic Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B duration 30 Minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.6 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.45 min; ESI-MS (+) m/z 962.3 (M+2H).

Analysis condition B: residence time = 2.74 min; ESI-MS (-) m/z 960.7 (M-2H).

ESI-HRMS (+) m/z: Calculated: 962.4824.

Preparation of Example 6403

Example 6403 was prepared following "General Synthesis Sequence A", wherein "Single Coupling Procedure" and "Double Coupling Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. Consolidation contains what you want The product was dissolved and dried by centrifugal evaporation. The yield of the product was 13.2 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.52 min; ESI-MS (+) m/z 965.3 (M+2H).

Analysis conditions B: residence time = 2.92 min; ESI-MS (+) m/z 965.0 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 6404

Example 6404 was prepared following "General Synthetic Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. Consolidation contains what you want The product was dissolved and dried by centrifugal evaporation. The yield of the product was 16.9 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z 979.1 (M+2H).

Analysis condition B: residence time = 2.87 min; ESI-MS (-) m/z 977.3 (M-2H).

Preparation of Example 6405

Example 6405 was prepared following "General Synthetic Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.5 mg, and The purity estimate was 98% based on LCMS analysis.

Analysis conditions A: residence time = 1.47 min; ESI-MS (+) m/z 966.2 (M+2H).

Analysis conditions B: residence time = 2.87 min; ESI-MS (+) m/z 965.4 (M+2H).

ESI-HRMS (+) m/z: Calculated: 965.4933.

Preparation of Example 6406

Example 6406 was prepared following "General Synthesis Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.7 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.32 min; ESI-MS (+) m/z 965.6 (M+2H).

Analysis conditions B: residence time = 2.70 min; ESI-MS (+) m/z 965.6 (M+2H).

ESI-HRMS (+) m/z: Calculated: 965.4933.

Preparation of Example 6407

Example 6407 was prepared following "General Synthetic Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.16 min; ESI-MS (+) m/z 953.1 (M+2H).

Analysis conditions B: residence time = 2.46 min; ESI-MS (-) m/z 951.1 (M-2H).

ESI-HRMS (+) m/z: Calculated: 95.

Preparation of Example 6408

Example 6408 was prepared following "General Synthetic Sequence A", wherein the "Single Coupling Procedure" and "Double Coupling Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.22 min; ESI-MS (+) m/z 952.8 (M+2H).

Analysis conditions B: residence time = 2.53 min; ESI-MS (+) m/z 953.1 (M+2H).

Preparation of Example 6409

Example 6409 was prepared following "General Synthesis Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.23 min; ESI-MS (+) m/z 945.7 (M+2H).

Analysis conditions B: residence time = 2.57 min; ESI-MS (+) m/z 945.2 (M+2H).

ESI-HRMS (+) m/z: Calculated: 940.

Preparation of Example 6410

Example 6410 was prepared following "General Synthesis Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.16 min; ESI-MS (+) m/z 953.0 (M+2H).

Analysis conditions B: residence time = 2.41 min; ESI-MS (+) m/z 953.2 (M+2H).

ESI-HRMS (+) m/z: Calculated: 95.

Preparation of Example 6411

Example 6411 was prepared following "General Synthetic Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-70% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.15 min; ESI-MS (-) m/z 977.0 (M- 2H).

Analysis conditions B: residence time = 2.36 min; ESI-MS (+) m/z 978.7 (M+2H).

ESI-HRMS (+) m/z: Calculated: 97.

Preparation of Example 6412

Example 6412 was prepared following "General Synthetic Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.17 min; ESI-MS (-) m/z 976.5 (M- 2H).

Analysis conditions B: residence time = 2.42 min; ESI-MS (+) m/z 978.3 (M+2H).

ESI-HRMS (+) m/z: Calculated: 97.

Preparation of Example 6415

Example 6415 was prepared following "General Synthesis Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95 : 5 methanol: water + 10 mM ammonium acetate; gradient: 40-85% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.1 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis condition A: residence time = 1.51 min; ESI-MS (+) m/z 975.8 (M+2H).

Analysis conditions B: residence time = 2.74 min; ESI-MS (+) m/z 977.9 (M+2H).

ESI-HRMS (+) m/z: Calculated: 976.9774.

Preparation of Example 6416

Example 6416 was prepared following "General Synthesis Sequence A", in which "Single Coupled Procedure" and "Double Coupled Procedure" of "Symphony Method A" were modified to use DMF (0.4 M, 1.0 mL) containing acetic anhydride (1.0 mL) + DIPEA. Instead of pure acetic anhydride (2.0 mL). The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol :water+ 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.5 mg and the purity was estimated to be 94% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (-) m/z 977.9 (M-2H).

Analysis conditions B: residence time = 2.77 min; ESI-MS (+) m/z 977.9 (M+2H).

Preparation of Example 7067

Example 7013 (3.0 mg, 1.631 umol) was dissolved in methanol (3 mL). The mixture was treated with Pd/C (1 mg), was bubbled under a hydrogen atmosphere (1 atm, balloon) and then stirred for 24 hours. LCMS showed consumption of unsaturated starting materials. The mixture was filtered to remove Pd/C. The solvent was removed to give Example 7067 (1.4 mg).

Analysis conditions A: residence time = 2.92 min; ESI-MS (+) m/z = 921.5 (M+2H).

ESI-HRMS (+) m/z calcd.

Examples 7068 through 7154 were prepared following the general synthetic sequence described for the preparation of Example 5001, which consisted of the following general procedure: "Prelude Method A: Resin Swelling Procedure", "Prelude Method A: Single Coupling Procedure", " Prelude Method A: Double Coupling Procedure", "Prelude Method A: Chloroacetyl chloride coupling procedure". The "Total Removal Protecting Method A" and "Circularization Method A" used in the preparation of Examples 7068 to 7154 were modified as described below. General Removal of Protecting Group Method A: Procedure for "Total Removal of Protective Group Method A" describes an experiment conducted on a scale of 0.100 mmol, where the scale is determined by the amount of Rink linker bound to the resin. The "removal of the protecting group solution" was prepared by combining trifluoroacetic acid (22 mL), phenol (1.325 g), water (1.25 mL) and triisopropyl decane (0.5 mL) in a 40 mL glass vial. The resin was removed from the reaction vessel and transferred to a 4 mL glass vial. A "Removal of the protecting group solution" (2.0 mL) was added to the vial. The mixture was mixed on an orbital shaker (175 RPM for 2 hours). The mixture was filtered through a funnel and the solid was washed with &lt;RTI ID=0.0&gt;&gt; To the combined filtrate was added Et ₂ O (15 mL). The mixture was vigorously mixed, followed by precipitation of a large amount of white solid. The mixture was centrifuged at 2000 RPM for 3 minutes, the solution was decanted from the solid and discarded. This process was repeated 3 times, then the solid was allowed to stand and air dried for 1 to 2 hours, then the crude peptide was obtained as a white solid. Cyclization Method A: The procedure of "Circularization Method A" describes an experiment conducted on a scale of 0.100 mmol. The crude solid was dissolved peptide MeCN: 0.1M NH aq ₄ OAc (15mL: 15mL), the solution was carefully adjusted to pH = 9.0 and then with aqueous NaOH (1.0M). The vial was capped and then mixed overnight at 175 RPM on an orbital shaker (about 18 hours). The reaction solution was concentrated and then the residue was dissolved in MeOH. This solution was subjected to reverse phase HPLC purification to give the desired cyclic peptide.

Final purity was determined using two of the four analytical LC/MS conditions.

Analytical conditions A: Column: Waters BEH C18, 2.0 x 50 mm, 1.7 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; temperature : 50 ° C; Gradient: 0% B, 0-100% B for 3 minutes, followed by 100 minutes at 100% B; flow rate: 1 mL/min; detection: 220 nm UV.

Analytical conditions B: Column: Waters BEH C18, 2.0 x 50 mm, 1.7 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; temperature : 50 ° C; Gradient: 0% B, 0-100% B lasts 3 minutes, then at 100% B was maintained for 0.5 minutes; flow rate: 0.5 mL/min; detection: 220 nm UV.

Analytical conditions C1: Column: Waters BEH C18, 2.0 x 50 mm, 1.7 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.05% TFA; mobile phase B: 95:5 acetonitrile: water + 0.05% TFA; : 50 ° C; Gradient: 0% B, 0-100% B for 3 minutes, followed by 100 minutes at 100% B; flow: 0.5 mL/min; detection: 220 nm UV.

Analysis conditions D1: Column: Waters BEH C18, 2.0 × 50 mm, 1.7 μm particles; mobile phase A: 5: 95 methanol: water + 0.05% TFA; mobile phase B: 95: 5 methanol: water + 0.05% TFA; : 50 ° C; Gradient: 0% B, 0-100% B for 3 minutes, followed by 100 minutes at 100% B; flow: 0.5 mL/min; detection: 220 nm UV.

Preparation of Example 7068

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 5 μm, 19 x 200 mm, mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: Water + 10 mM ammonium acetate; gradient: 40-80% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 1.9 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.84 min; ESI-MS (+) m/z = 940.6 (M+2H).

Analysis conditions B: residence time = 3.06 min; ESI-MS (+) m/z = 940.5 (M+2H).

Preparation of Example 7069

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.85 min; ESI-MS (+) m/z = 957.5 (M+2H).

Analysis condition B: residence time = 3.07 min; ESI-MS (+) m/z = 957.5 (M+2H).

ESI-HRMS (+) m/z calc. 955.9383, calc. 955.9382 (M+2H).

Preparation of Example 7070

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.8 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.86 min; ESI-MS (+) m/z = 929.9 (M+2H).

Analysis conditions B: residence time = 3.06 min; ESI-MS (+) m/z = 929.7 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7071

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.7 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.91 min; ESI-MS (+) m/z = 955.4 (M+2H).

Analysis condition B: residence time = 3.02 min; ESI-MS (+) m/z = 955.2 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7072

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.3 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.66 min; ESI-MS (+) m/z = 98.2 (M+2H).

Analysis condition B: residence time = 2.80 min; ESI-MS (+) m/z =978.3 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7073

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19×mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.7 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition A: retention time = 1.50min; ESI-MS (+ ) m / z = 937.4 (M + 2H).

Analysis condition B: residence time = 2.59 min; ESI-MS (+) m/z = 937.7 (M+2H).

Preparation of Example 7075

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 25-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.2 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.50 min; ESI-MS (+) m/z = 963.2 (M+2H).

Analysis condition B: residence time = 2.59 min; ESI-MS (+) m/z = 963.6 (M+2H).

ESI-HRMS (+) m/z calc., s.

Preparation of Example 7077

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19×mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.43 min; ESI-MS (+) m/z = 975.2 (M+2H).

Analytical condition B: residence time = 2.58 min; ESI-MS (+) m/z = 95.7 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7078

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.4 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.63 min; ESI-MS (+) m/z = 968.6 (M+2H).

Analysis condition B: residence time = 2.77 min; ESI-MS (+) m/z = 968.6 (M+2H).

ESI-HRMS (+) m/z calcd., s.

Preparation of Example 7079

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 25-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.65 min; ESI-MS (+) m/z = 979.6 (M+2H).

Analysis condition B: residence time = 2.89 min; ESI-MS (+) m/z = 979.4 (M+2H).

ESI-HRMS (+) m/z calc. 978. 592.

Preparation of Example 7080

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 38.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.86 min; ESI-MS (+) m/z = 973.1 (M+2H).

Analysis condition B: residence time = 3.03 min; ESI-MS (+) m/z = 973.6 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7081

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.59 min; ESI-MS (+) m/z = 971.8 (M+2H).

Analysis condition B: residence time = 2.82 min; ESI-MS (+) m/z = 971.6 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7082

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.75 min; ESI-MS (+) m/z = 965.2 (M+2H).

Analysis condition B: residence time = 2.95 min; ESI-MS (+) m/z = 965.5 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7083

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.89 min; ESI-MS (+) m/z = 951.5 (M+2H).

Analysis condition B: residence time = 3.00 min; ESI-MS (+) m/z = 951.3 (M+2H).

ESI-HRMS(+) m/z calculated 950.9826, experimental value 950.9820 (M+2H).

Preparation of Example 7084

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 5 μm, 19 x 200 mm, mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: Water + 10 mM ammonium acetate; Gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.79 min; ESI-MS (+) m/z = 923.6 (M+2H).

Analysis conditions B: residence time = 2.92 min; ESI-MS (+) m/z = 923.6 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7085

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.60 min; ESI-MS (+) m/z = 977.6 (M+2H).

Analysis condition B: residence time = 2.77 min; ESI-MS (+) m/z = 977.8 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7086

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.2 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis condition A: residence time = 1.74 min; ESI-MS (+) m/z = 971.0 (M+2H).

Analysis condition B: residence time = 2.89 min; ESI-MS (+) m/z = 971.1 (M+2H).

ESI-HRMS (+) m/z calc., s.

Preparation of Example 7087

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-85% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.2 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.64 min; ESI-MS (+) m/z = 957.6 (M+2H).

Analysis condition B: residence time = 2.93 min; ESI-MS (+) m/z = 957.0 (M+2H).

ESI-HRMS (+) m/z calcd. 95.. 992.

Preparation of Example 7088

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 25-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 33.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.63 min; ESI-MS (+) m/z = 931.3 (M+2H).

Analysis condition B: residence time = 3.01 min; ESI-MS (+) m/z = 931.8 (M+2H).

Preparation of Example 7089

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 44.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.50 min; ESI-MS (+) m/z = 930.7 (M+2H).

Analysis condition B: residence time = 2.93 min; ESI-MS (+) m/z = 930.6 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7090

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-85% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 38.9 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.58 min; ESI-MS (+) m/z = 949.9 (M+2H).

Analysis condition B: residence time = 2.81 min; ESI-MS (+) m/z = 949.6 (M+2H).

ESI-HRMS (+) m/z calc. </RTI>

Preparation of Example 7091

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 41.3 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.59 min; ESI-MS (+) m/z = 931.1 (M+2H).

Analysis condition B: residence time = 2.81 min; ESI-MS (+) m/z = 931.4 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7092

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-85% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 49.5 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.47 min; ESI-MS (+) m/z = 924.1 (M+2H).

Analysis conditions B: residence time = 2.78 min; ESI-MS (+) m/z = 924.2 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7093

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-85% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.55 min ; ESI-MS (+) m/z = 956.6 (M+2H).

Analysis condition B: residence time = 2.91 min; ESI-MS (+) m/z = 956.2 (M+2H).

ESI-HRMS (+) m/z calc. 955.4983, calc. 955.4978 (M+2H).

Preparation of Example 7094

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 39.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.52 min; ESI-MS (+) m/z = 956.6 (M+2H).

Analysis condition B: residence time = 2.93 min; ESI-MS (+) m/z = 956.7 (M+2H).

Preparation of Example 7095

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 35.6 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition C1: residence time = 1.31 min; ESI-MS (+) m/z = 948.7 (M+2H).

Analysis condition D1: residence time = 3.48 min; ESI-MS (+) m/z = 948.8 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7096

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.8 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.59 min; ESI-MS (+) m/z = 931.5 (M+2H).

Analysis condition B: residence time = 3.03 min; ESI-MS (+) m/z = 931.1 (M+2H).

Preparation of Example 7097

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-100% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.64 min; ESI-MS (+) m/z = 964.3 (M+2H).

Analysis conditions B: residence time = 2.96 min; ESI-MS (+) m/z = 964.2 (M+2H).

ESI-HRMS (+) m/z calcd.

Preparation of Example 7098

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-85% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 31.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.52 min; ESI-MS (+) m/z = 957.6 (M+2H).

Analysis conditions B: residence time = 2.79 min; ESI-MS (+) m/z = 957.0 (M+2H).

ESI-HRMS (+) m/z calc. 956.4642.

Preparation of Example 7099

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 60-100% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.65 min; ESI-MS (+) m/z = 964.5 (M+2H).

Analysis condition B: residence time = 2.93 min; ESI-MS (+) m/z = 964.5 (M+2H).

ESI-HRMS (+) m/z calc., s.

Preparation of Example 7100

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 31.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.70 min; ESI-MS (+) m/z = 938.7 (M+2H).

Analysis condition B: residence time = 2.99 min; ESI-MS (+) m/z = 938.9 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7101

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 32.4 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.66 min; ESI-MS (+) m/z = 939.1 (M+2H).

Analysis condition B: residence time = 2.98 min; ESI-MS (+) m/z = 939.1 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7102

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 39.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.58 min; ESI-MS (+) m/z = 938.9 (M+2H).

Analysis conditions B: residence time = 2.84 min; ESI-MS (+) m/z = 938.6 (M+2H).

ESI-HRMS (+) m/z calc. </RTI></RTI></RTI></RTI></RTI>

Preparation of Example 7103

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.4 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition C1: residence time = 1.28 min; ESI-MS (+) m/z = 923.4 (M+2H).

Analysis condition D1: residence time = 2.77 min; ESI-MS (+) m/z = 923.3 (M+2H).

ESI-HRMS (+) m/z calcd.

Preparation of Example 7104

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-100% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 40.6 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.60 min; ESI-MS (+) m/z = 957.0 (M+2H).

Analysis condition B: residence time = 2.91 min; ESI-MS (+) m/z = 957.1 (M+2H).

ESI-HRMS (+) m/z calc. 956.4642.

Preparation of Example 7105

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.55 min ; ESI-MS (+) m/z = 931.6 (M+2H).

Analysis conditions B: residence time = 2.87 min; ESI-MS (+) m/z = 931.8 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7106

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 33.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.68 min; ESI-MS (+) m/z = 964.0 (M+2H).

Analysis conditions B: residence time = 2.99 min; ESI-MS (+) m/z = 964.2 (M+2H).

ESI-HRMS (+) m/z calcd.

Preparation of Example 7107

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 36.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.45 min; ESI-MS (+) m/z = 956.5 (M+2H).

Analysis condition B: residence time = 2.74 min; ESI-MS (+) m/z = 956.6 (M+2H).

ESI-HRMS (+) m/z calcd.

Preparation of Example 7108

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 41.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.64 min; ESI-MS (+) m/z = 938.2 (M+2H).

Analysis conditions B: residence time = 2.94 min; ESI-MS (+) m/z = 938.5 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7109

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 36.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.59 min; ESI-MS (+) m/z = 964.0 (M+2H).

Analysis condition B: residence time = 2.83 min; ESI-MS (+) m/z = 964.1 (M+2H).

ESI-HRMS (+) m/z calcd.

Preparation of Example 7110

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 41.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z = 964.6 (M+2H).

Analysis condition B: residence time = 2.74 min; ESI-MS (+) m/z = 964.6 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7111

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and repurified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; Phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.52 min; ESI-MS (+) m/z = 957.3 (M+2H).

Analysis condition B: residence time = 2.80 min; ESI-MS (+) m/z = 957.0 (M+2H).

ESI-HRMS (+) m/z calc. 956.4642.

Preparation of Example 7112

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 43.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.47 min; ESI-MS (+) m/z = 931.6 (M+2H).

Analysis condition B: residence time = 2.80 min; ESI-MS (+) m/z = 931.5 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7113

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 34.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.58 min; ESI-MS (+) m/z = 939.0 (M+2H).

Analysis condition B: residence time = 2.91 min; ESI-MS (+) m/z = 939.1 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7114

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z = 939.7 (M+2H).

Analysis conditions B: residence time = 2.48 min; ESI-MS (+) m/z = 939.7 (M+2H).

ESI-HRMS(+) m/z calculated value 938.4404, experimental value 938.4375 (M+2H)

Preparation of Example 7115

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.53 min; ESI-MS (+) m/z = 971.1 (M+2H).

Analysis conditions B: residence time = 2.57 min; ESI-MS (+) m/z = 971.0 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7116

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.55 min ; ESI-MS (+) m/z = 938.8 (M+2H).

Analysis conditions B: residence time = 2.54 min; ESI-MS (+) m/z = 938.5 (M+2H).

ESI-HRMS(+) m/z calculated value 933.7665, experimental value 937.9649 (M+2H)

Preparation of Example 7117

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-80% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.8 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition A: retention time = 1.41min; ESI-MS (+ ) m / z = 945.7 (M + 2H).

Analysis condition B: residence time = 2.66 min; ESI-MS (+) m/z = 945.8 (M+2H).

ESI-HRMS(+) m/z calc. 945.4300, mp.

Preparation of Example 7118

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-85% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 35.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.69 min; ESI-MS (+) m/z = 963.4 (M+2H).

Analysis condition B: residence time = 3.25 min; ESI-MS (+) m/z = 963.4 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7119

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 52.7 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.67 min; ESI-MS (+) m/z = 963.9 (M+2H).

Analysis condition B: retention time = 3.31min; ESI-MS (+ ) m / z = 964.0 (M + 2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7120

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-85% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 48.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: retention time = 1.51min; ESI-MS (+ ) m / z = 956.4 (M + 2H).

Analysis condition B: residence time = 3.03 min; ESI-MS (+) m/z = 956.3 (M+2H).

ESI-HRMS (+) m/z calcd. 955.9903, calc. 955.9896 (M+2H).

Preparation of Example 7121

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.66 min; ESI-MS (+) m/z = 937.9 (M+2H).

Analysis condition B: residence time = 3.26 min; ESI-MS (+) m/z = 937.9 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7122

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 46.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.71 min; ESI-MS (+) m/z = 938.4 (M+2H).

Analysis condition B: residence time = 3.35 min; ESI-MS (+) m/z = 938.4 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7123

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-85% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 39.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.44 min; ESI-MS (+) m/z = 930.9 (M+2H).

Analysis condition B: residence time = 3.01 min; ESI-MS (+) m/z = 931.0 (M+2H).

ESI-HRMS(+) m/z calculated value 930.4849, experimental value 930.4835 (M+2H).

Preparation of Example 7124

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z = 971.4 (M+2H).

Analysis condition B: residence time = 2.95 min; ESI-MS (+) m/z = 971.4 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7125

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 39.6 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.51 min; ESI-MS (+) m/z = 971.9 (M+2H).

Analysis condition B: residence time = 2.95 min; ESI-MS (+) m/z = 971.9 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7126

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.8 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.35 min; ESI-MS (+) m/z = 964.7 (M+2H).

Analysis conditions B: residence time = 2.85 min; ESI-MS (+) m/z = 964.9 (M+2H).

ESI-HRMS (+) m/z calcd.

Preparation of Example 7127

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 37.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.45 min; ESI-MS (+) m/z = 945.4 (M+2H).

Analysis condition B: residence time = 2.95 min; ESI-MS (+) m/z = 945.4 (M+2H).

ESI-HRMS (+) m/z calcd. 940. 592.

Preparation of Example 7128

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-100% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: retention time = 1.74min; ESI-MS (+ ) m / z = 963.4 (M + 2H).

Analysis conditions B: residence time = 3.29 min; ESI-MS (+) m/z = 963.4 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7129

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 37.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.65 min; ESI-MS (+) m/z = 938.5 (M+2H).

Analysis condition B: residence time = 2.98 min; ESI-MS (+) m/z = 938.4 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7130

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-80% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 44.0 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.42 min; ESI-MS (+) m/z = 946.1 (M+2H).

Analysis conditions B: residence time = 2.64 min; ESI-MS (+) m/z = 946.0 (M+2H).

ESI-HRMS (+) m/z calcd: 945.448.

Preparation of Example 7131

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.40 min; ESI-MS (+) m/z = 971.3 (M+2H).

Analysis condition B: residence time = 2.69 min; ESI-MS (+) m/z = 971.2 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7132

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 32.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.28 min; ESI-MS (+) m/z = 964.3 (M+2H).

Analysis condition B: residence time = 2.58 min; ESI-MS (+) m/z = 964.2 (M+2H).

ESI-HRMS (+) m/z calcd., s.

Preparation of Example 7133

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.1 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.45 min; ESI-MS (+) m/z = 946.2 (M+2H).

Analysis condition B: residence time = 2.72 min; ESI-MS (+) m/z = 946.2 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7134

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 99.9 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.59 min; ESI-MS (+) m/z = 931.8 (M+2H).

Analysis conditions B: residence time = 2.59 min; ESI-MS (+) m/z = 931.2 (M+2H).

ESI-HRMS (+) m/z calc .

Preparation of Example 7135

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 32.3 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition A: residence time = 1.94 min; ESI-MS (+) m/z = 948.8 (M+2H), 957.5 (M+H+NH ₄ )

Analysis condition B: residence time = 2.90 min; ESI-MS (+) m/z = 949.8 (M+2H), 957.5 (M+H+NH ₄ )

ESI-HRMS (+) m/z calc. 948.4.

Preparation of Example 7136

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 41.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.84 min; ESI-MS (+) m/z = 924.0 (M+2H).

Analysis condition B: residence time = 3.39 min; ESI-MS (+) m/z = 924.0 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7137

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 38.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.84 min; ESI-MS (+) m/z = 936.0 (M+2H).

Analysis condition B: residence time = 3.36 min; ESI-MS (+) m/z = 936.0 (M+2H).

ESI-HRMS (+) m/z calc .

Preparation of Example 7138

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 34.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: retention time = 1.89min; ESI-MS (+ ) m / z = 911.0 (M + 2H).

Analysis condition B: residence time = 3.40 min; ESI-MS (+) m/z = 911.0 (M+2H).

ESI-HRMS(+) m/z calculated 910.4272, experimental value 910.4266 (M+2H)

Preparation of Example 7139

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 33.2 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.76 min; ESI-MS (+) m/z = 943.0 (M+2H).

Analysis condition B: residence time = 2.90 min; ESI-MS (+) m/z = 943.7 (M+2H).

(+) M / z calcd ESI-HRMS 942.4485, Found 942.4482 (M + 2H).

Preparation of Example 7140

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.33 min; ESI-MS (+) m/z = 968.0 (M+2H).

Analysis conditions B: residence time = 2.70 min; ESI-MS (+) m/z = 968.0 (M+2H).

(+) M / z calcd ESI-HRMS 967.4431, Found 967.4406 (M + 2H).

Preparation of Example 7141

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.4 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.70 min; ESI-MS (+) m/z = 978.7 (M+2H).

Analysis condition B: retention time = 3.27min; ESI-MS (+ ) m / z = 978.6 (M + 2H).

(+) M / z calcd ESI-HRMS 977.4694, Found 977.4683 (M + 2H).

Preparation of Example 7142

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.3 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.71 min; ESI-MS (+) m/z = 952.8 (M+2H).

Analysis condition B: residence time = 2.31 min; ESI-MS (+) m/z = 952.9 (M+2H).

Preparation of Example 7143

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.47 min; ESI-MS (+) m/z = 965.2 (M+2H).

Analysis conditions B: residence time = 2.78 min; ESI-MS (+) m/z = 965.8 (M+2H).

ESI-HRMS (+) m/z calc., s.

Preparation of Example 7144

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.4 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.67 min; ESI-MS (+) m/z = 940.8 (M+2H).

Analysis condition B: residence time = 2.22 min; ESI-MS (+) m/z = 940.7 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7145

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.1 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.60 min; ESI-MS (+) m/z = 971.4 (M+2H).

Analysis condition B: residence time = 3.22 min; ESI-MS (+) m/z = 971.8 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7146

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 38.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.43 min; ESI-MS (+) m/z = 967.7 (M+2H).

Analysis condition B: residence time = 2.74 min; ESI-MS (+) m/z = 967.6 (M+2H).

ESI-HRMS (+) m/z calc.

Preparation of Example 7147

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 36.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.44 min; ESI-MS (+) m/z = 993.7 (M+2H).

Analysis condition B: residence time = 3.08 min; ESI-MS (+) m/z = 993.6 (M+2H).

ESI-HRMS (+) m/z calcd. s.

Preparation of Example 7148

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.47 min; ESI-MS (+) m/z = 967.4 (M+2H).

Analysis condition B: residence time = 3.15 min; ESI-MS (+) m/z = 967.4 (M+2H).

Preparation of Example 7151

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.69 min; ESI-MS (+) m/z = 951.7 (M+2H).

Analysis condition B: residence time = 2.93 min; ESI-MS (+) m/z = 951.2 (M+2H).

Preparation of Example 7152

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.3 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis condition A: residence time = 1.45 min; ESI-MS (+) m/z = 946.6 (M+ 2H).

Analysis condition B: residence time = 2.49 min; ESI-MS (+) m/z = 946.9 (M+2H).

Preparation of Example 7153

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation.

The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation.

The yield of the product was 6.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.55 min; ESI-MS (+) m/z = 999.5 (M+2H).

Analysis conditions B: residence time = 2.87 min; ESI-MS (+) m/z = 999.8 (M+2H).

Preparation of Example 7154

The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation.

The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation.

The yield of the product was 1.2 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.67 min; ESI-MS (+) m/z = 999.7 (M+2H).

Analysis conditions B: residence time = 3.03 min; ESI-MS (+) m/z = 999.8 (M+2H).

General program for SymphonyX Method D Analysis data:

Mass spectrometry: "ESI-MS (+)" indicates electrospray ionization mass spectrometry in positive ion mode; "ESI-MS (-)" indicates electrospray ionization mass spectrometry in negative ion mode; "ESI-HRMS (+)" indicates High-resolution electrospray ionization mass spectrometry in positive ion mode; "ESI-HRMS(-)" represents high-resolution electrospray ionization mass spectrometry in negative ion mode. The detected quality is reported after the " m/z " unit name. Compounds with an accurate mass greater than 1000 are often detected as ions with double or triple charges.

Analysis condition A:

Column: Waters BEH C18, 2.0×50 mm, 1.7 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; temperature: 50 ° C; Gradient: 0% B, 0-100% B lasted 3 minutes, then held at 100% B for 0.5 minutes; flow rate: 1 mL/min; detection: 220 nm UV.

Analysis condition B:

Column: Waters BEH C18, 2.0×50 mm, 1.7 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; temperature: 50 ° C; Gradient: 0% B, 0-100% B lasted 3 minutes, then held at 100% B for 0.5 minutes; flow rate: 0.5 mL/min; detection: 220 nm UV.

Universal coupling procedure D:

All operations were performed on a Symphony X Peptide Synthesizer (Protein Technologies) according to automated operation. Unless otherwise noted, all procedures were carried out in a 10 mL polypropylene tube equipped with a bottom frit; in the case of a reaction scale exceeding 0.100 mmol, a 40 mL polypropylene tube fitted with a bottom frit was used. The tube was connected to the Symphony X peptide synthesizer via the bottom and top of the tube. DMF and DCM can be added via the top of the tube, which is equally flushed along the side of the tube. The remaining reagent is added to the resin via the top of the tube and up through the frit. All solutions were removed via the bottom of the tube. "Stirred periodically" described in brief pulses N ₂ gas through the bottom frit; pulse duration from about 5 seconds and occur once every 30 seconds. The solution of chloroacetic chloride in DMF was used within 24 hours of preparation. The amino acid solution is usually used up to three weeks after preparation. The HATU solution was used within 5 days of preparation. DMF = dimethylformamide; HATU = hexafluorophosphate 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3 - Oxide; DIPEA = diisopropylethylamine; Rink resin: 4-((2,4-dimethoxyphenyl)(Fmocamino)methyl)phenoxymethylpolystyrene. Sieber amide resin: 9-Fmoc-amino group - -3-yloxy-methyl, a binding polymer.

The procedure for "Symphony X Method D" describes an experiment conducted on a scale of 0.100 mmol. All procedures can be scaled up by more than 0.100 mmol by adjusting the volume by a multiple of this size. Prior to amino acid coupling, all peptide synthesis sequences begin with a resin swelling procedure, described below as "resin swelling procedure." The N-terminal coupling of an amino acid with a primary amine uses the following "single coupling procedure". The N-terminal coupling of the amino acid with the secondary amine uses the following "double coupling procedure". The coupling of chloroacetamidine chloride to the N-terminus of the peptide is described by the "chloroethonium chloride coupling procedure" detailed below.

Resin swelling procedure:

A resin (0.100 mmol) was added to a 10 mL polypropylene solid phase reaction vessel. The resin was washed (swelled) three times as follows: DMF (2.0 mL) was added to the reaction vessel, and then the mixture was periodically stirred for 10 minutes, and then the solvent was discharged through a glass frit.

Single coupling procedure:

Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to a reaction vessel containing the resin from the previous step. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. The resin was washed six times in succession as follows: DFI (2.0 mL) was added via the top of the vessel and the resulting mixture was weekly for each wash. The mixture was stirred for 30 seconds, and then the solution was discharged through the glass frit. Amino acid (0.2 M in DMF, 1.0 mL, 2 eq) was added to the reaction vessel, followed by HATU (0.2M in DMF, 1.0 mL, 2 eq), and finally DIPEA (0.4M in DMF, 1.0 mL) , 4eq). The mixture was stirred periodically for 15 minutes, and then the reaction solution was discharged through a frit. The resin was washed four times as follows: DFI (2.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 30 seconds, after which the solution was discharged through the frit. Acetic anhydride (2.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 10 minutes and then the solution was discharged through a frit. The resin was washed four times continuously as follows: DFI (2.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 90 seconds, after which the solution was discharged through the frit. The resulting resin was used directly in the next step.

Single coupling 1 hour program:

Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to a reaction vessel containing the resin from the previous step. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. The resin was washed six times in succession as follows: DMF (2.0 mL) was added via the top of the vessel for each wash and the resulting mixture was stirred periodically for 30 seconds before the solution was discharged through the frit. Amino acid (0.2 M in DMF, 1.0 mL, 2 eq) was added to the reaction vessel, followed by HATU (0.2M in DMF, 1.0 mL, 2 eq), and finally DIPEA (0.4M in DMF, 1.0 mL) , 4eq). The mixture was stirred periodically for 60 minutes, and then the reaction solution was discharged through a glass frit. The resin was washed four times as follows: DFI (2.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 30 seconds, after which the solution was discharged through the frit. Acetic anhydride (2.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 10 minutes and then the solution was discharged through a frit. The resin was washed four times continuously as follows: DFI (2.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 90 seconds, after which the solution was discharged through the frit. The resulting resin was used directly in the next step.

Double coupling procedure

Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to a reaction vessel containing the resin from the previous step. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. The resin was washed six times in succession as follows: DMF (2.0 mL) was added via the top of the vessel for each wash and the resulting mixture was stirred periodically for 30 seconds before the solution was discharged through the frit. Amino acid (0.2 M in DMF, 1.0 mL, 2 eq) was added to the reaction vessel, followed by HATU (0.2M in DMF, 1.0 mL, 2 eq), and finally DIPEA (0.4M in DMF, 1.0 mL) , 4eq). The mixture was stirred periodically for 15 minutes, and then the reaction solution was discharged through a frit. The resin was washed twice as follows: at each wash, DMF (2.0 mL) was added via the top of the vessel and the resulting mixture was stirred periodically for 30 seconds, then the solution was drained through the frit. Amino acid (0.2 M in DMF, 1.0 mL, 2 eq) was added to the reaction vessel, followed by HATU (0.2M in DMF, 1.0 mL, 2 eq), and finally DIPEA (0.4M in DMF, 1.0 mL) , 4eq). The mixture was stirred periodically for 15 minutes, and then the reaction solution was discharged through a frit. The resin was washed twice as follows: at each wash, DMF (2.0 mL) was added via the top of the vessel and the resulting mixture was stirred periodically for 30 seconds, then the solution was drained through the frit. Acetic anhydride (2.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 10 minutes and then the solution was discharged through a frit. The resin was washed four times continuously as follows: DFI (2.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 90 seconds, after which the solution was discharged through the frit. The resulting resin was used directly in the next step.

Double coupling 6 hour program:

Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to a reaction vessel containing the resin from the previous step. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. The resin is washed continuously as follows Times: At each wash, DMF (2.0 mL) was added via the top of the vessel and the resulting mixture was stirred periodically for 30 seconds before the solution was drained through the frit. Amino acid (0.2 M in DMF, 1.0 mL, 2 eq) was added to the reaction vessel, followed by HATU (0.2M in DMF, 1.0 mL, 2 eq), and finally DIPEA (0.4M in DMF, 1.0 mL) , 4eq). The mixture was stirred periodically for 6 hours, and then the reaction solution was discharged through a glass frit. The resin was washed twice as follows: at each wash, DMF (2.0 mL) was added via the top of the vessel and the resulting mixture was stirred periodically for 30 seconds, then the solution was drained through the frit. Amino acid (0.2 M in DMF, 1.0 mL, 2 eq) was added to the reaction vessel, followed by HATU (0.2M in DMF, 1.0 mL, 2 eq), and finally DIPEA (0.4M in DMF, 1.0 mL) , 4eq). The mixture was stirred periodically for 6 hours, and then the reaction solution was discharged through a glass frit. The resin was washed twice as follows: at each wash, DMF (2.0 mL) was added via the top of the vessel and the resulting mixture was stirred periodically for 30 seconds, then the solution was drained through the frit. Acetic anhydride (2.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 10 minutes and then the solution was discharged through a frit. The resin was washed four times continuously as follows: DFI (2.0 mL) was added via the top of the vessel with each wash and the resulting mixture was periodically stirred for 90 seconds, after which the solution was discharged through the frit. The resulting resin was used directly in the next step.

Chloroacetyl chloride coupling procedure:

Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to a reaction vessel containing the resin from the previous step. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. Piperidine: DMF (20: 80 v/v, 2.0 mL) was added to the reaction vessel. The mixture was stirred periodically for 3 minutes and then the solution was discharged through a frit. The resin was washed six times in succession as follows: DMF (2.0 mL) was added via the top of the vessel for each wash and the resulting mixture was stirred periodically for 30 seconds before the solution was discharged through the frit. DIPEA (0.8 M in DMF, 3.0 mL, 24 eq) was added to the reaction mixture, followed by chloroethyl chloride (0.8M in DMF, 1.65 mL, 13.2 eq). The mixture was stirred periodically for 30 minutes, and then the reaction solution was discharged through a glass frit. The resin was washed three times in succession as follows: DFI (2.0 mL) was added to the vessel at the top of each wash and the resulting mixture was periodically stirred for 90 seconds, after which the solution was discharged through the frit. The resin was washed four times in the following manner: On each wash, CH ₂ Cl ₂ (2.0 mL) was added to the vessel at the top and the resulting mixture was stirred periodically for 90 seconds, after which the solution was discharged through the glass frit. The resulting resin was placed under a stream of N ₂ for 15 minutes.

Removal of protecting group method D:

All operations are performed manually unless otherwise noted. The procedure for "Removal of Protecting Group Method D" describes an experiment conducted on a scale of 0.100 mmol. The procedure can be expanded by more than 0.100 mmol by adjusting the volume in multiples of this size. A "removal protecting group solution" was prepared by combining trifluoroacetic acid (38 mL), DTT (1 g), and triisopropyl decane (1 mL) in a 40 mL glass vial. The resin was removed from the reaction vessel and transferred to a 4 mL glass vial. A "Removal of the protecting group solution" (2.0 mL) was added to the vial. The mixture was vigorously mixed in an oscillator (1000 RPM for 15-30 minutes). The mixture was filtered through a 0.2 micron syringe filter and the solid was extracted with a "deprotection solution" (1.0 mL). To a 24 mL tube containing the combined filtrate was added Et ₂ O (15 mL). The mixture was vigorously mixed, followed by precipitation of a large amount of white solid. The mixture was centrifuged for 3 minutes, then the solution was decanted from the solid and discarded. The solid was suspended in Et ₂ O (20 mL); the mixture was then centrifuged for 3 min; and the solution was decanted from the solid and discarded. Finally, the solid was suspended in Et ₂ O (20 mL); the mixture was centrifuged for 3 minutes; and the solution was decanted from the solid and discarded to give the crude peptide as a white to pale white solid.

Cyclization method D:

All operations are performed manually unless otherwise noted. The procedure of "Circulation Method D" describes an experiment conducted on a scale of 0.100 mmol. The procedure can be expanded by more than 0.100 mmol by adjusting the volume in multiples of this size. The crude solid was dissolved peptide MeCN: 0.1M NH ₄ OAc aq (1: 1) until a total volume of 18 to 22mL, and then the solution was carefully adjusted to pH = 8.5-9.0 with aqueous NaOH (1.0M). The solution was then allowed to stand for 12 to 18 hours without agitation. The reaction solution was concentrated and the residue was dissolved in MeOH:MeOH (1:1). This solution was subjected to reverse phase HPLC purification to give the desired cyclic peptide.

General synthetic order D:

"General Synthesis Sequence D" describes the general sequence of procedures used to obtain the cyclic peptides described herein. Rink or Sieber resin was added to a 10 mL polypropylene solid phase reaction vessel, and the reaction vessel was placed on a Symphony X peptide synthesizer. "Universal coupling procedure D": Follow the resin swelling procedure. Then, on the Symphony X peptide synthesizer, follow the "single coupling procedure" (if the N-terminus of the resin-binding peptide is a primary amine) or the "double coupling procedure" (if the N-terminus of the resin-binding peptide is a secondary amine), one in sequence A series of amino acid couplings. The "single coupling for 1 hour" procedure was used to give the amino acid (S)-2-(((9H-indol-9-yl)methoxy)carbonyl)amino)-3-(3,4,5- Trifluorophenyl)propionic acid is coupled to the resin. If the proline is coupled to the resin, the "double coupling 6 hour procedure" is used. Following the chloroethyl hydrazine coupling procedure; followed by removal of the protecting group method D; followed by cyclization method D.

9000 series

Examples 9115 through 9187 were prepared following "General Synthesis Sequence A". Examples 9188 through 9196, examples 9213 through 9224, examples 9241 through 9288, and examples 9318 through 9374 were prepared following "Universal Synthesis Sequence D."

Preparation of Example 9115

The crude material of Example 9115 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.9 mg and the purity was estimated to be 91% according to LCMS analysis.

Analysis conditions A: residence time = 1.37 min; ESI-MS (+) m/z 910.1 (M+2H).

Analysis condition B: residence time = 2.42 min; ESI-MS (+) m/z 910.1 (M+2H).

Preparation of Example 9116

The crude material of Example 9116 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.54 min; ESI-MS (+) m/z 918.3 (M+2H).

Analysis condition B: retention time = 2.64min; ESI-MS (+ ) m / z 918.6 (M + 2H).

Preparation of Example 9117

The crude material of Example 9117 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.9 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.33 min; ESI-MS (+) m/z 913.7 (M+2H).

Analysis condition B: residence time = 2.36 min; ESI-MS (+) m/z 914.1 (M+2H).

Preparation of Example 9118

The crude material of Example 9118 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.43 min; ESI-MS (+) m/z 922.6 (M+2H).

Analysis conditions B: residence time = 2.58 min; ESI-MS (+) m/z 922.6 (M+2H).

Preparation of Example 9119

The crude material of Example 9119 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.28 min; ESI-MS (+) m/z 922.2 (M+2H).

Analysis conditions B: residence time = 2.34 min; ESI-MS (+) m/z 922.2 (M+2H).

Preparation of Example 9120

The crude material of Example 9120 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.45 min; ESI-MS (+) m/z 930.7 (M+2H).

Analysis conditions B: residence time = 2.56 min; ESI-MS (+) m/z 930.7 (M+2H).

Preparation of Example 9121

The crude material of Example 9121 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.8 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.39 min; ESI-MS (+) m/z 916.9 (M+2H).

Analysis conditions B: residence time = 2.45 min; ESI-MS (+) m/z 917.1 (M+2H).

Preparation of Example 9122

The crude material of Example 9122 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.52 min; ESI-MS (+) m/z 925.6 (M+2H).

Analysis condition B: retention time = 2.65min; ESI-MS (+ ) m / z 925.5 (M + 2H).

Preparation of Example 9123

The crude material of Example 9231 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.38 min; ESI-MS (+) m/z 925.2 (M+2H).

Analysis condition B: retention time = 2.40min; ESI-MS (+ ) m / z 925.3 (M + 2H).

Preparation of Example 9124

The crude material of Example 9124 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.6 mg and the purity was estimated to be 93% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z 933.9 (M+2H).

Analysis condition B: retention time = 2.62min; ESI-MS (+ ) m / z 933.7 (M + 2H).

Preparation of Example 9125

The crude material of Example 9125 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.3 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition A: retention time = 1.47min; ESI-MS (+ ) m / z 942.1 (M + 2H).

Analysis condition B: retention time = 2.59min; ESI-MS (+ ) m / z 942.2 (M + 2H).

Preparation of Example 9126

The crude material of Example 9126 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.4 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.59 min; ESI-MS (+) m/z 950.7 (M+2H).

Analysis conditions B: residence time = 2.75 min; ESI-MS (+) m/z 950.7 (M+2H).

Preparation of Example 9127

The crude material of Example 9127 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.5 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition A: retention time = 1.53min; ESI-MS (+ ) m / z 938.5 (M + 2H).

Analysis conditions B: residence time = 2.65 min; ESI-MS (+) m/z 938.6 (M+2H).

Preparation of Example 9128

The crude material of Example 9128 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, then held at 100 wB for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.5 mg and the purity was estimated to be 93% according to LCMS analysis.

Analysis conditions A: residence time = 1.50 min; ESI-MS (+) m/z 941.5 (M+2H).

Analysis conditions B: residence time = 2.59 min; ESI-MS (+) m/z 941.8 (M+2H).

Preparation of Example 9129

The crude material of Example 9129 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.6 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis condition A: residence time = 1.67 min; ESI-MS (-) m/z 949.9 (M-2H).

Analysis condition B: retention time = 2.79min; ESI-MS (+ ) m / z 952.1 (M + 2H).

Preparation of Example 9130

The crude material of Example 9130 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.3 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.50 min; ESI-MS (+) m/z 951.7 (M+2H).

Analysis conditions B: residence time = 2.56 min; ESI-MS (+) m/z 951.6 (M+2H).

Preparation of Example 9131

The crude material of Example 9131 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.8 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.63 min; ESI-MS (+) m/z 960.0 (M+2H).

Analysis conditions B: residence time = 2.79 min; ESI-MS (+) m/z 960.2 (M+2H).

Preparation of Example 9132

The crude material of Example 9132 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.58 min; ESI-MS (-) m/z 892.1 (M-2H).

Analysis conditions B: residence time = 2.67 min; ESI-MS (+) m/z 894.1 (M+2H).

Preparation of Example 9133

The crude material of Example 9133 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.76 min; ESI-MS (+) m/z 902.6 (M+2H).

Analysis conditions B: residence time = 2.88 min; ESI-MS (+) m/z 902.6 (M+2H).

Preparation of Example 9134

The crude material of Example 9134 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.67 min; ESI-MS (+) m/z 902.0 (M+2H).

Analysis conditions B: residence time = 2.65 min; ESI-MS (+) m/z 902.0 (M+2H).

Preparation of Example 9135

The crude material 9135 of the example was subjected to a single compound purification group for purification and analysis. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.73 min; ESI-MS (+) m/z 910.9 (M+2H).

Analysis condition B: retention time = 2.90min; ESI-MS (+ ) m / z 910.6 (M + 2H).

Preparation of Example 9136

The crude material of Example 9136 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.7 mg and the purity was estimated to be 92% according to LCMS analysis.

Analysis conditions A: residence time = 1.35 min; ESI-MS (+) m/z 915.7 (M+2H).

Analysis conditions B: residence time = 2.42 min; ESI-MS (+) m/z 915.6 (M+2H).

Preparation of Example 9137

The crude material of Example 9137 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 5-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.53 min; ESI-MS (+) m/z 924.0 (M+2H).

Analysis conditions B: residence time = 2.65 min; ESI-MS (+) m/z 924.3 (M+2H).

Preparation of Example 9138

The crude material of Example 9138 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.32 min; ESI-MS (+) m/z 924.1 (M+2H).

Analysis condition B: residence time = 2.35 min; ESI-MS (+) m/z 924.0 (M+2H).

Preparation of Example 9139

The crude material of Example 9139 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 10-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95 : 5 methanol: water + 10 mM ammonium acetate; gradient: 30-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.51 min; ESI-MS (+) m/z 932.4 (M+2H).

Analysis conditions B: residence time = 2.63 min; ESI-MS (+) m/z 932.2 (M+2H).

Preparation of Example 9140

The crude material of Example 9140 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.8 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.60 min; ESI-MS (+) m/z 902.2 (M+2H).

Analysis conditions B: residence time = 2.60 min; ESI-MS (+) m/z 902.0 (M+2H).

Preparation of Example 9141

The crude material of Example 9141 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.9 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.76 min; ESI-MS (+) m/z 910.5 (M+2H).

Analysis conditions B: residence time = 2.86 min; ESI-MS (+) m/z 910.6 (M+2H).

Preparation of Example 9142

The crude material of Example 9142 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.8 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.57 min; ESI-MS (+) m/z 910.1 (M+2H).

Analysis conditions B: residence time = 2.63 min; ESI-MS (+) m/z 910.0 (M+2H).

Preparation of Example 9143

The crude material of Example 9143 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.3 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.68 min; ESI-MS (+) m/z 918.9 (M+2H).

Analysis conditions B: residence time = 2.84 min; ESI-MS (+) m/z 918.5 (M+2H).

Preparation of Example 9144

The crude material of Example 9144 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.8 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.54 min; ESI-MS (+) m/z 948.5 (M+2H).

Analysis conditions B: residence time = 2.60 min; ESI-MS (+) m/z 948.2 (M+2H).

Preparation of Example 9145

The crude material of Example 9145 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.66 min; ESI-MS (+) m/z 956.5 (M+2H).

Analysis conditions B: residence time = 2.83 min; ESI-MS (+) m/z 956.6 (M+2H).

Preparation of Example 9146

The crude material of Example 9146 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z 956.0 (M+2H).

Analysis condition B: residence time = 2.57 min; ESI-MS (+) m/z 956.1 (M+2H).

Preparation of Example 9147

The crude material of Example 9147 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.7 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.61 min; ESI-MS (+) m/z 965.0 (M+2H).

Analysis conditions B: residence time = 2.78 min; ESI-MS (+) m/z 964.7 (M+2H).

Preparation of Example 9148

The crude material of Example 9148 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.31 min; ESI-MS (+) m/z 931.2 (M+2H).

Analysis conditions B: residence time = 2.50 min; ESI-MS (+) m/z 931.0 (M+2H).

Preparation of Example 9149

The crude material of Example 9149 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 25-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.37 min; ESI-MS (+) m/z 939.7 (M+2H).

Analysis conditions B: residence time = 2.62 min; ESI-MS (+) m/z 939.1 (M+2H).

Preparation of Example 9150

The crude material of Example 9150 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.63 min; ESI-MS (+) m/z 946.6 (M+2H).

Analysis conditions B: residence time = 2.80 min; ESI-MS (+) m/z 946.6 (M+2H).

Preparation of Example 9155

The crude material of Example 9155 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.54 min; ESI-MS (+) m/z 915.8 (M+2H).

Analysis condition B: residence time = 2.63 min; ESI-MS (+) m/z 916.1 (M+2H).

Preparation of Example 9156

The crude material of Example 9156 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.68 min; ESI-MS (-) m/z 922.1 (M-2H).

Analysis conditions B: residence time = 2.83 min; ESI-MS (+) m/z 924.0 (M+2H).

Preparation of Example 9157

The crude material of Example 9157 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.8 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.53 min; ESI-MS (+) m/z 923.8 (M+2H).

Analysis conditions B: residence time = 2.60 min; ESI-MS (+) m/z 923.6 (M+2H).

Preparation of Example 9158

The crude material of Example 9158 was subjected to a single compound purification group for purification and analysis. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.9 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.64 min; ESI-MS (+) m/z 932.2 (M+2H).

Analysis conditions B: residence time = 2.80 min; ESI-MS (+) m/z 932.0 (M+2H).

Preparation of Example 9159

The crude material of Example 9159 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.74 min; ESI-MS (+) m/z 913.5 (M+2H).

Analysis conditions B: residence time = 2.80 min; ESI-MS (+) m/z 915.1 (M+2H).

Preparation of Example 9160

The crude material of Example 9160 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 35-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.89 min; ESI-MS (+) m/z 923.3 (M+2H).

Analysis conditions B: residence time = 3.04 min; ESI-MS (+) m/z 923.7 (M+2H).

Preparation of Example 9161

The crude material of Example 9161 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.2 mg and the purity was estimated to be 93% according to LCMS analysis.

Analysis conditions A: residence time = 1.70 min; ESI-MS (+) m/z 923.1 (M+2H).

Analysis conditions B: residence time = 2.75 min; ESI-MS (+) m/z 923.1 (M+2H).

Preparation of Example 9162

The crude material of Example 9162 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.9 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.53 min; ESI-MS (+) m/z 908.7 (M+2H).

Analysis conditions B: residence time = 2.63 min; ESI-MS (+) m/z 908.8 (M+2H).

Preparation of Example 9163

The crude material of Example 9163 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 25-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.3 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.66 min; ESI-MS (+) m/z 917.2 (M+2H).

Analysis condition B: residence time = 2.84 min; ESI-MS (+) m/z 917.3 (M+2H).

Preparation of Example 9164

The crude material of Example 9164 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.0 mg and the purity was estimated to be 93% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z 916.9 (M+2H).

Analysis conditions B: residence time = 2.60 min; ESI-MS (+) m/z 916.6 (M+2H).

Preparation of Example 9165

The crude material of Example 9165 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 34.9 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.66 min; ESI-MS (+) m/z 925.1 (M+2H).

Analysis condition B: residence time = 2.81 min; ESI-MS (+) m/z 925.1 (M+2H).

Preparation of Example 9166

The crude material of Example 9166 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.7 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.58 min; ESI-MS (+) m/z 950.2 (M+2H).

Analysis conditions B: residence time = 2.72 min; ESI-MS (+) m/z 949.9 (M+2H).

Preparation of Example 9167

The crude material of Example 9167 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 30-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.7 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.44 min; ESI-MS (+) m/z 949.9 (M+2H).

Analysis condition B: residence time = 2.61 min; ESI-MS (+) m/z 950.0 (M+2H).

Preparation of Example 9168

The crude material of Example 9168 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.76 min; ESI-MS (+) m/z 922.8 (M+2H).

Analysis condition B: residence time = 2.47 min; ESI-MS (+) m/z 923.1 (M+2H).

Preparation of Example 9169

The crude material of Example 9169 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.45 min; ESI-MS (+) m/z 930.90 (M+2H).

Analysis conditions B: residence time = 2.88 min; ESI-MS (+) m/z 930.95 (M+2H).

Preparation of Example 9170

The crude material of Example 9170 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.0 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.40 min; ESI-MS (+) m/z 934.65 (M+2H).

Analysis conditions B: residence time = 2.83 min; ESI-MS (+) m/z 934.80 (M+2H).

Preparation of Example 9171

The crude material of Example 9171 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.36 min; ESI-MS (+) m/z 942.85 (M+2H).

Analysis conditions B: residence time = 2.78 min; ESI-MS (+) m/z 942.95 (M+2H).

Preparation of Example 9172

The crude material of Example 9172 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.35 min; ESI-MS (+) m/z 936.60 (M+2H).

Analysis condition B: residence time = 1.93 min; ESI-MS (+) m/z 936.80 (M+2H).

Preparation of Example 9173

The crude material of Example 9173 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.28 min; ESI-MS (+) m/z 944.60 (M+2H).

Analysis conditions B: residence time = 1.87 min; ESI-MS (+) m/z 944.60 (M+2H).

Preparation of Example 9174

The crude material of Example 9174 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.4 mg and the purity was estimated to be 92% according to LCMS analysis.

Analysis conditions A: residence time = 1.26 min; ESI-MS (+) m/z 948.55 (M+2H).

Analysis conditions B: residence time = 2.63 min; ESI-MS (+) m/z 948.30 (M+2H).

Preparation of Example 9175

The crude material of Example 9175 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 0-40% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.22 min; ESI-MS (+) m/z 956.80 (M+2H).

Analysis condition B: residence time = 1.76 min; ESI-MS (+) m/z 956.70 (M+2H).

Preparation of Example 9176

The crude material of Example 9176 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.9 mg and the purity was estimated to be 92% according to LCMS analysis.

Analysis conditions A: residence time = 1.31 min; ESI-MS (+) m/z 922.40 (M+2H).

Analysis conditions B: residence time = 2.78 min; ESI-MS (+) m/z 922.40 (M+2H).

Preparation of Example 9177

The crude material of Example 9177 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.44 min; ESI-MS (+) m/z 930.05 (M+2H).

Analysis conditions B: residence time = 2.69 min; ESI-MS (+) m/z 930.05 (M+2H).

Preparation of Example 9178

The crude material of Example 9178 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.41 min; ESI-MS (+) m/z 934.15 (M+2H).

Analysis conditions B: residence time = 2.89 min; ESI-MS (+) m/z 934.20 (M+2H).

Preparation of Example 9179

The crude material of Example 9179 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.1 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.37 min; ESI-MS (+) m/z 942.20 (M+2H).

Analysis conditions B: residence time = 2.70 min; ESI-MS (+) m/z 942.05 (M+2H).

Preparation of Example 9180

The crude material of Example 9180 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.52 min; ESI-MS (+) m/z 929.20 (M+2H).

Analysis conditions B: residence time = 2.99 min; ESI-MS (+) m/z 929.25 (M+2H).

Preparation of Example 9181

The crude material of Example 9181 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.9 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.49 min; ESI-MS (+) m/z 937.20 (M+2H).

Analysis conditions B: residence time = 2.99 min; ESI-MS (+) m/z 937.40 (M+2H).

Preparation of Example 9182

The crude material of Example 9182 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.7 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.53 min; ESI-MS (+) m/z 941.45 (M+2H).

Analysis condition B: residence time = 2.99 min; ESI-MS (+) m/z 941.40 (M+2H).

Preparation of Example 9183

The crude material of Example 9183 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.9 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.49 min; ESI-MS (+) m/z 949.35 (M+2H).

Analysis condition B: residence time = 2.91 min; ESI-MS (+) m/z 949.45 (M+2H).

Preparation of Example 9184

The crude material of Example 9184 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.45 min; ESI-MS (+) m/z 929.90 (M+2H).

Analysis condition B: residence time = 2.92 min; ESI-MS (+) m/z 929.95 (M+2H).

Preparation of Example 9185

The crude material of Example 9185 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.0 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.42 min; ESI-MS (+) m/z 937.85 (M+2H).

Analysis condition B: residence time = 2.88 min; ESI-MS (+) m/z 937.90 (M+2H).

Preparation of Example 9186

The crude material of Example 9186 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.28 min; ESI-MS (+) m/z 941.65 (M+2H).

Analysis condition B: residence time = 2.53 min; ESI-MS (+) m/z 941.65 (M+2H).

Preparation of Example 9187

The crude material of Example 9187 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 30-75% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.24 min; ESI-MS (+) m/z 949.65 (M+2H).

Analysis conditions B: residence time = 2.49 min; ESI-MS (+) m/z 949.65 (M+2H).

Preparation of Example 9188

The crude material of Example 9188 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19×mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 1.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 2.10 min; ESI-MS (+) m/z 919.8 (M+2H).

Analysis conditions B: residence time = 2.74 min; ESI-MS (+) m/z 919.2 (M+2H).

Preparation of Example 9189

The crude material of Example 9189 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.2 mg and the purity was estimated to be 93% according to LCMS analysis.

Analysis conditions A: residence time = 2.07 min; ESI-MS (+) m/z 925.9 (M+2H).

Analysis conditions B: residence time = 2.73 min; ESI-MS (+) m/z 926.0 (M+2H).

Preparation of Example 9190

The crude material of Example 9190 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.4 mg, and the purity was estimated to be 10 analytical conditions A: retention time = 2.13 min; ESI-MS (+) m/z 918.3 (M+2H).

Analysis conditions B: residence time = 2.85 min; ESI-MS (+) m/z 918.5 (M+2H).

Preparation of Example 9191

The crude material of Example 9191 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 60-100% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 27.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 2.02 min; ESI-MS (+) m/z 923.6 (M+2H).

Analysis condition B: residence time = 2.83 min; ESI-MS (+) m/z 923.6 (M+2H).

Preparation of Example 9192

The crude material of Example 9192 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 30-70% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.92 min; ESI-MS (+) m/z 911.70 (M+2H).

Analysis condition B: residence time = 2.53 min; ESI-MS (+) m/z 911.45 (M+2H).

Preparation of Example 9193

The crude material of Example 9193 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 60-100% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min.

The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.82 min; ESI-MS (+) m/z 919.3 (M+2H).

Analysis conditions B: residence time = 2.76 min; ESI-MS (+) m/z 919.7 (M+2H).

Preparation of Example 9194

The crude material of Example 9194 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.8 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.81 min; ESI-MS (+) m/z 926.3 (M+2H).

Analysis conditions B: residence time = 2.73 min; ESI-MS (+) m/z 926.7 (M+2H).

Preparation of Example 9195

The crude material of Example 9195 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.2 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.919.2 min; ESI-MS (+) m/z 919.2 (M+2H).

Analysis conditions B: residence time = 2.76 min; ESI-MS (+) m/z 919.1 (M+2H).

Preparation of Example 9196

The crude material of Example 9196 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 2.04 min; ESI-MS (+) m/z 926.4 (M+2H).

Analysis conditions B: residence time = 2.72 min; ESI-MS (+) m/z 926.5 (M+2H).

Preparation of Example 9213

The crude material of Example 9213 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 34.2 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.80 min; ESI-MS (+) m/z 916.55 (M+2H).

Analysis condition B: residence time = 3.40 min; ESI-MS (+) m/z 916.60 (M+2H).

Preparation of Example 9214

The crude material of Example 9214 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.2 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.87 min; ESI-MS (+) m/z 923.60 (M+2H).

Analysis conditions B: residence time = 2.55 min; ESI-MS (+) m/z 923.55 (M+2H).

Preparation of Example 9215

The crude material of Example 9215 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.76 min; ESI-MS (+) m/z 899.15 (M+2H).

Analysis condition B: residence time = 2.44 min; ESI-MS (+) m/z 899.05 (M+2H).

Preparation of Example 9216

The crude material of Example 9216 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 54.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.81 min; ESI-MS (+) m/z 906.20 (M+2H).

Analysis conditions B: residence time = 3.42 min; ESI-MS (+) m/z 906.10 (M+2H).

Preparation of Example 9217

The crude material of Example 9217 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 60.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.73 min; ESI-MS (+) m/z 899.55 (M+2H).

Analysis condition B: residence time = 3.41 min; ESI-MS (+) m/z 899.65 (M+2H).

Preparation of Example 9218

The crude material of Example 9218 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 44.1 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.59 min; ESI-MS (+) m/z 906.60 (M+2H).

Analysis condition B: residence time = 3.43 min; ESI-MS (+) m/z 906.70 (M+2H).

Preparation of Example 9219

The crude material of Example 9219 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.68 min; ESI-MS (+) m/z 917.95 (M+2H).

Analysis conditions B: residence time = 3.27 min; ESI-MS (+) m/z 917.95 (M+2H).

Preparation of Example 9220

The crude material of Example 9220 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 36.2 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.73 min; ESI-MS (+) m/z 924.95 (M+2H).

Analysis conditions B: residence time = 3.25 min; ESI-MS (+) m/z 924.90 (M+2H).

Preparation of Example 9221

The crude material of Example 9221 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 43.3 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.63 min; ESI-MS (+) m/z 918.40 (M+2H).

Analysis conditions B: residence time = 3.27 min; ESI-MS (+) m/z 918.40 (M+2H).

Preparation of Example 9222

The crude material of Example 9222 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 33.2 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.66 min; ESI-MS (+) m/z 925.50 (M+2H).

Analysis condition B: residence time = 3.35 min; ESI-MS (+) m/z 925.40 (M+2H).

Preparation of Example 9223

The crude material of Example 9223 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 36.0 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.61 min; ESI-MS (+) m/z 900.40 (M+2H).

Analysis condition B: residence time = 3.18 min; ESI-MS (+) m/z 900.40 (M+2H).

Preparation of Example 9224

The crude material of Example 9224 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 38.5 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.68 min; ESI-MS (+) m/z 907.25 (M+2H).

Analysis condition B: residence time = 3.28 min ; ESI-MS (+) m/z 907.40 (M+2H).

Preparation of Examples 9225 and 9226

Examples 9225 and 9226 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9225 was 27.4 mg and the purity was estimated to be 96% based on LCMS analysis. The yield of Example 9226 was 24.9 mg and the purity was estimated to be 96% based on LCMS analysis.

Example 9225

Analysis conditions A: residence time = 1.77 min; ESI-MS (+) m/z 966.45 (M+2H).

Analysis condition B: residence time = 3.34 min; ESI-MS (+) m/z 966.45 (M+2H).

Example 9266

Analysis conditions A: residence time = 1.93 min; ESI-MS (+) m/z 966.40 (M+2H).

Analysis condition B: residence time = 3.50 min; ESI-MS (+) m/z 966.85 (M+2H).

Preparation of Examples 9227 and 9228

Examples 9227 and 9228 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9227 was 39.1 mg and the purity was estimated to be 98% according to LCMS analysis. The yield of Example 9228 was 21.6 mg and the purity was estimated to be 95% based on LCMS analysis.

Example 9227

Analysis conditions A: residence time = 1.70 min; ESI-MS (+) m/z 958.90 (M+2H).

Analysis condition B: residence time = 3.27 min; ESI-MS (+) m/z 958.90 (M+2H).

Example 9228

Analysis conditions A: residence time = 1.79 min; ESI-MS (+) m/z 958.90 (M+2H).

Analysis condition B: residence time = 3.36 min; ESI-MS (+) m/z 958.95 (M+2H).

Preparation of Examples 9229 and 9230

Examples 9229 and 930 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9229 was 25.1 mg and the purity was estimated to be 98% according to LCMS analysis. The yield of Example 9229 was 9270 mg of 9230, and the purity was estimated to be 98% according to LCMS analysis.

Example 9229

Analysis conditions A: residence time = 1.76 min; ESI-MS (+) m/z 965.95 (M+2H).

Analysis condition B: residence time = 3.35 min; ESI-MS (+) m/z 965.95 (M+2H).

Example 9230

Analysis conditions A: residence time = 1.98 min; ESI-MS (+) m/z 965.85 (M+2H).

Analysis conditions B: residence time = 3.47 min; ESI-MS (+) m/z 965.55 (M+2H).

Preparation of Examples 9231 and 9232

Examples 9231 and 9232 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B over 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9231 was 24.9 mg and the purity was estimated to be 100% according to LCMS analysis. The yield of Example 9232 was 16.0 mg and the purity was estimated to be 96% based on LCMS analysis.

Example 9231

Analysis conditions A: residence time = 1.67 min; ESI-MS (+) m/z 959.40 (M+2H).

Analysis conditions B: residence time = 3.25 min; ESI-MS (+) m/z 959.40 (M+2H).

Example 9232

Analysis conditions A: residence time = 1.78 min; ESI-MS (+) m/z 959.40 (M+2H).

Analysis conditions B: residence time = 3.32 min; ESI-MS (+) m/z 959.35 (M+2H).

Preparation of Examples 9233 and 9234

Examples 9233 and 9234 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9233 was 13.3 mg and the purity was estimated to be 93% according to LCMS analysis. The yield of Example 9234 was 11.1 mg and the purity was estimated to be 94% according to LCMS analysis.

Example 9233

Analysis conditions A: residence time = 1.54 min; ESI-MS (+) m/z 960.0 (M+2H).

Analysis condition B: residence time = 2.70 min; ESI-MS (+) m/z 960.0 (M+2H).

Example 9234

Analysis conditions A: residence time = 1.59 min; ESI-MS (+) m/z 960.1 (M+2H).

Analysis conditions B: residence time = 2.78 min; ESI-MS (+) m/z 960.4 (M+2H).

Preparation of Examples 9235 and 9236

Examples 9235 and 9236 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9235 was 13.0 mg and the purity was estimated to be 90% according to LCMS analysis. The yield of the example 9236 was 10.3 mg and the purity was estimated to be 92% according to LCMS analysis.

Example 9235

Analysis conditions A: residence time = 1.62 min; ESI-MS (+) m/z 966.95 (M+2H).

Analysis conditions B: residence time = 3.16 min; ESI-MS (+) m/z 966.90 (M+2H).

Example 9236

Analysis conditions A: residence time = 1.79 min; ESI-MS (+) m/z 966.75 (M+2H).

Analysis conditions B: residence time = 3.37 min; ESI-MS (+) m/z 966.95 (M+2H).

Preparation of Examples 9237 and 9238

Examples 9237 and 9238 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9237 was 20.4 mg and the purity was estimated to be 96% according to LCMS analysis. The yield of Example 9238 was 20.5 mg and the purity was estimated to be 97% according to LCMS analysis.

Example 9237

Analysis condition A: residence time = 1.52 min; ESI-MS (+) m/z 960.40 (M+2H).

Analysis condition B: residence time = 3.05 min; ESI-MS (+) m/z 960.40 (M+2H).

Example 9238

Analysis conditions A: residence time = 1.62 min; ESI-MS (+) m/z 960.40 (M+2H).

Analysis condition B: residence time = 3.13 min; ESI-MS (+) m/z 960.40 (M+2H).

Preparation of Examples 9239 and 9240

Examples 9239 and 9240 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9239 was 22.7 mg and the purity was estimated to be 97% according to LCMS analysis. The yield of Example 9240 was 16.0 mg and the purity was estimated to be 97% according to LCMS analysis.

Example 9239

Analysis conditions A: residence time = 1.59 min; ESI-MS (+) m/z 967.45 (M+2H).

Analysis condition B: residence time = 3.13 min; ESI-MS (+) m/z 967.45 (M+2H).

Example 9240

Analysis conditions A: residence time = 1.75 min; ESI-MS (+) m/z 967.40 (M+2H).

Analysis conditions B: residence time = 3.29 min; ESI-MS (+) m/z 967.40 (M+2H).

Preparation of Example 9241

The crude material of Example 9241 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 25-65% B lasts 30 minutes, then 100% B for 5 minutes; flow rate: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.65 min; ESI-MS (-) m/z 908.5 (M-2H).

Analysis condition B: residence time = 2.86 min; ESI-MS (+) m/z 908.1 (M-2H).

Preparation of Example 9242

The crude material of Example 9242 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 25-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.6 mg and was classified according to LCMS. The purity was estimated to be 97%.

Analysis conditions A: residence time = 1.82 min; ESI-MS (+) m/z 916.20 (M+2H).

Analysis condition B: residence time = 2.47 min; ESI-MS (+) m/z 916.45 (M-2H).

Preparation of Example 9243

The crude material of Example 9243 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.3 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition A: residence time = 1.66 min; ESI-MS (-) m/z 909.0 (M-2H).

Analysis condition B: residence time = 3.30 min; ESI-MS (+) m/z 910.10 (M+2H).

Preparation of Example 9244

The crude material of Example 9244 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition A: residence time = 1.69 min; ESI-MS (-) m/z 916.0 (M-2H).

Analysis conditions B: residence time = 3.43 min; ESI-MS (+) m/z 917.05 (M+2H).

Preparation of Example 9245

The crude material of Example 9245 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.58 min; ESI-MS (-) m/z 891.4 (M-2H).

Analysis conditions B: residence time = 3.04 min; ESI-MS (+) m/z 892.55 (M+2H).

Preparation of Example 9246

The crude material of Example 9246 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition B: residence time = 3.14 min; ESI-MS (+) m/z 899.55 (M+2H).

Preparation of Example 9247

The crude material of Example 9427 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.4 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions B: residence time = 3.12 min; ESI-MS (+) m/z 893.00 (M+2H).

Preparation of Example 9248

The crude material of Example 9248 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions B: residence time = 3.26 min; ESI-MS (+) m/z 900.0 (M+2H).

Preparation of Example 9249

The crude material of Example 9249 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 20 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.6 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis condition B: residence time = 3.38 min; ESI-MS (+) m/z 916.55 (M+2H).

Preparation of Example 9250

The crude material of Example 9250 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19×mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 41.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.85 min; ESI-MS (-) m/z 922.5 (M-2H).

Analysis condition B: residence time = 3.48 min; ESI-MS (+) m/z 923.45 (M+2H).

Preparation of Example 9251

The crude material of Example 9251 was purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; Phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 38.1 mg, and the purity was estimated to be 94% according to LCMS analysis. Analysis conditions A: retention time = 1. min; ESI-MS (+) m/z (M+2H).

Analysis condition A: residence time = 1.71 min; ESI-MS (-) m/z 916.6 (M-2H).

Analysis conditions B: residence time = 2.39 min; ESI-MS (+) m/z 917.05 (M+2H).

Preparation of Example 9252

The crude material of Example 9252 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 33.1 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition A: residence time = 1.76 min; ESI-MS (-) m/z 923.0 (M-2H).

Analysis conditions B: residence time = 3.06 min; ESI-MS (+) m/z 925.7 (M+2H).

Preparation of Example 9253

The crude material of Example 9253 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 47.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.63 min; ESI-MS (-) m/z 898.6 (M-2H).

Analysis condition B: residence time = 3.19 min; ESI-MS (+) m/z 899.50 (M+2H).

Preparation of Example 9254

The crude material of Example 9254 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 48.9 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition A: residence time = 1.64 min; ESI-MS (-) m/z 905.8 (M-2H).

Analysis condition B: residence time = 2. min; ESI-MS (+) m/z 906.55 (M+2H).

Preparation of Example 9255

The crude material of Example 9255 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 52.2 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.63 min; ESI-MS (+) m/z 899.2 (M+2H).

Analysis conditions B: residence time = 3.23 min; ESI-MS (+) m/z 900.05 (M+2H).

Preparation of Example 9256

The crude material of Example 9256 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 53.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.69 min; ESI-MS (+) m/z 906.8 (M+2H).

Analysis condition B: residence time = 3.40 min; ESI-MS (+) m/z 907.00 (M+2H).

Preparation of Example 9257

The crude material of Example 9257 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.0 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 2.88 min; ESI-MS (+) m/z 915.7 (M+2H).

Analysis condition B: residence time = 3.28 min ; ESI-MS (+) m/z 916.55 (M+2H).

Preparation of Example 9258

The crude material of Example 9258 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.7 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.77 min; ESI-MS (+) m/z 922.8 (M+2H).

Analysis conditions B: residence time = 2.76 min; ESI-MS (+) m/z 923.60 (M+2H).

Preparation of Example 9259

The crude material of Example 9259 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.75 min; ESI-MS (-) m/z 923.0 (M-2H).

Analysis condition B: residence time = 3.47 min; ESI-MS (+) m/z 924.05 (M+2H).

Preparation of Example 9260

The crude material of Example 9260 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition B: residence time = 3.08 min; ESI-MS (-) m/z 899.55 (M-2H).

Preparation of Example 9261

The crude material of Example 9261 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.67 min; ESI-MS (+) m/z 905.5 (M+2H).

Analysis condition B: residence time = 3.19 min; ESI-MS (+) m/z 906.55 (M+2H).

Preparation of Example 9262

The crude material of Example 9262 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions B: residence time = 3.18 min; ESI-MS (+) m/z 899.9 (M+2H).

Preparation of Example 9263

The crude material of Example 9263 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 33.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.63 min; ESI-MS (+) m/z 905.6 (M+2H).

Analysis condition B: residence time = 3.31 min; ESI-MS (+) m/z 907.10 (M+2H).

Preparation of Example 9264

The crude material of Example 9264 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 60-100% B for 25 minutes, followed by 100% B for 10 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.9 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.74 min; ESI-MS (+) m/z 968.25 (M+2H).

Analysis condition B: residence time = 2.38 min; ESI-MS (+) m/z 968.45 (M+2H).

Preparation of Example 9265

The crude material of Example 9265 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 60-100% B for 25 minutes, followed by 100% B for 10 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.6 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.68 min; ESI-MS (+) m/z 969.15 (M+2H).

Analysis condition B: residence time = 3.32 min; ESI-MS (+) m/z 969.05 (M+2H).

Preparation of Example 9266

The crude material of Example 9266 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 60-100% B for 25 minutes, followed by 100% B for 10 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.68 min; ESI-MS (-) m/z 950.2 (M-2H).

Analysis conditions B: residence time = 2.85 min; ESI-MS (+) m/z 951.8 (M+2H).

Preparation of Example 9267

The crude material of Example 9267 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.1 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.67 min; ESI-MS (+) m/z 951.50 (M+2H).

Analysis conditions B: residence time = 3.26 min; ESI-MS (+) m/z 951.55 (M+2H).

Preparation of Example 9268

The crude material of Example 9268 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.2 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.61 min; ESI-MS (+) m/z 951.85 (M+2H).

Analysis conditions B: residence time = 3.18 min; ESI-MS (+) m/z 951.95 (M+2H).

Preparation of Example 9269

The crude material of Example 9269 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.40 min; ESI-MS (+) m/z 945.35 (M+2H).

Analysis conditions B: residence time = 2.74 min; ESI-MS (+) m/z 945.35 (M+2H).

Preparation of Example 9270

The crude material of Example 9270 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.32 min; ESI-MS (+) m/z 980.35 (M+2H).

Analysis conditions B: residence time = 2.74 min; ESI-MS (+) m/z 980.45 (M+2H).

Preparation of Example 9271

The crude material of Example 9271 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 39.6 mg and the purity was estimated to be 94% according to LCMS analysis.

Analysis conditions A: residence time = 1.52 min; ESI-MS (+) m/z 973.40 (M+2H).

Analysis condition B: residence time = 3.12 min; ESI-MS (+) m/z 973.35 (M+2H).

Preparation of Example 9272

The crude material of Example 9272 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.8 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.82 min; ESI-MS (+) m/z 973.50 (M+2H).

Analysis condition B: residence time = 3.08 min; ESI-MS (+) m/z 973.45 (M+2H).

Preparation of Example 9273

The crude material of Example 9273 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 25-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 48.5 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.60 min; ESI-MS (+) m/z 967.70 (M+2H).

Analysis conditions B: residence time = 2.99 min; ESI-MS (+) m/z 967.70 (M+2H).

Preparation of Example 9274

The crude material of Example 9274 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 25-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 46.8 mg and the purity was estimated to be 94% according to LCMS analysis.

Analysis conditions A: residence time = 1.60 min; ESI-MS (+) m/z 968.80 (M+2H).

Analysis condition B: residence time = 3.04 min; ESI-MS (+) m/z 968.20 (M+2H).

Preparation of Example 9275

The crude material of Example 9275 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 50.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.55 min ; ESI-MS (+) m/z 950.70 (M+2H).

Analysis condition B: residence time = 2.87 min; ESI-MS (+) m/z 950.70 (M+2H).

Preparation of Example 9276

The crude material of Example 9276 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.9 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1. min; ESI-MS (+) m/z 951.20 (M+2H).

Analysis conditions B: residence time = 2.96 min; ESI-MS (+) m/z 951.15 (M+2H).

Preparation of Example 9277

The crude material of Example 9277 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 43.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.50 min; ESI-MS (+) m/z 941.70 (M+2H).

Analysis conditions B: residence time = 2.75 min; ESI-MS (+) m/z 941.70 (M+2H).

Preparation of Example 9278

The crude material of Example 9278 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.50 min; ESI-MS (+) m/z 942.20 (M+2H).

Analysis conditions B: residence time = 2.91 min; ESI-MS (+) m/z 942.20 (M+2H).

Preparation of Example 9279

The crude material of Example 9279 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 52.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.63 min; ESI-MS (+) m/z 942.3 (M+2H).

Analysis conditions B: residence time = 2.86 min; ESI-MS (+) m/z 942.8 (M+2H).

Preparation of Example 9280

The crude material of Example 9280 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 42.2 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.68 min; ESI-MS (+) m/z 951.3 (M+2H).

Analysis conditions B: residence time = 2.76 min; ESI-MS (+) m/z 951.6 (M+2H).

Preparation of Example 9281

The crude material of Example 9281 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 56.9 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.51 min; ESI-MS (+) m/z 942.25 (M+2H).

Analysis conditions B: residence time = 2.93 min; ESI-MS (+) m/z 942.25 (M+2H).

Preparation of Example 9282

The crude material of Example 9282 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 150 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.4 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.60 min; ESI-MS (+) m/z 951.4 (M+2H).

Analysis conditions B: residence time = 2.78 min; ESI-MS (+) m/z 951.4 (M+2H).

Preparation of Example 9283

The crude material of Example 9283 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 52.5 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.84 min; ESI-MS (+) m/z 967.65 (M+2H).

Analysis condition B: residence time = 3.42 min; ESI-MS (+) m/z 967.90 (M+2H).

Preparation of Example 9284

The crude material of Example 9284 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 39.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.72 min; ESI-MS (+) m/z 967.90 (M+2H).

Analysis conditions B: residence time = 3.16 min; ESI-MS (+) m/z 968.25 (M+2H).

Preparation of Example 9285

The crude material 9285 of the example was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 50.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.58 min; ESI-MS (+) m/z 942.75 (M+2H).

Analysis conditions B: residence time = 2.89 min; ESI-MS (+) m/z 942.75 (M+2H).

Preparation of Example 9286

The crude material of Example 9286 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 59.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.75 min; ESI-MS (-) m/z 941.10 (M-2H).

Analysis condition B: residence time = 3.33 min; ESI-MS (+) m/z 943.35 (M+2H).

Preparation of Example 9287

The crude material of Example 9287 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 25-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 47.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.85 min; ESI-MS (+) m/z 960.40 (M+2H).

Analysis conditions B: residence time = 2.50 min; ESI-MS (+) m/z 959.50 (M+2H).

Preparation of Example 9288

The crude material of Example 9288 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 33.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.81 min; ESI-MS (-) m/z 958.10 (M-2H).

Analysis condition B: residence time = 3.48 min; ESI-MS (+) m/z 960.15 (M+2H).

Preparation of Example 9289

Example 9289 was prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.8 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.72 min; ESI-MS (-) m/z 931.75 (M-2H).

Analysis conditions B: residence time = 2.33 min; ESI-MS (+) m/z 933.60 (M+2H).

Preparation of Examples 9290 and 9291

Examples 9290 and 9291 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9290 was 4.8 mg and the purity was estimated to be 100% according to LCMS analysis. The yield of Example 9291 was 9.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Example 9290

Analysis conditions A: residence time = 1.76 min; ESI-MS (-) m/z 938.90 (M-2H).

Analysis conditions B: residence time = 3.36 min; ESI-MS (+) m/z 941.10 (M+2H).

Example 9291

Analysis conditions A: residence time = 1.79 min; ESI-MS (+) m/z 938.10 (M-2H).

Preparation of Examples 9292 and 9293

Examples 9292 and 9293 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. Consolidation contains what you want The product was dissolved and dried by centrifugal evaporation. The yield of Example 9292 was 3.1 mg and the purity was estimated to be 100% according to LCMS analysis. The yield of Example 9293 was 2.9 mg and its purity was estimated to be 100% according to LCMS analysis.

Example 9292

Analysis conditions B: residence time = 3.39 min; ESI-MS (+) m/z 934.25 (M+2H).

Example 9293

Analysis condition B: residence time = 3.39 min; ESI-MS (+) m/z 934.15 (M+2H).

Preparation of Examples 9294 and 9295

Examples 9294 and 9295 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: waters xbridge c-18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B :95:5 acetonitrile: water +0.1% three Fluoroacetic acid; Gradient: 20-60% B for 30 minutes followed by 100 minutes at 100% B; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9294 was 5.2 mg and the purity was estimated to be 100% according to LCMS analysis. The yield of the example 9295 was 3.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Example 9294

Analysis conditions A: residence time = 1.82 min; ESI-MS (+) m/z 940.95 (M+2H).

Example 9295

Analysis conditions A: residence time = 1.89 min; ESI-MS (+) m/z 940.90 (M+2H).

Preparation of Examples 9296 and 9297

Examples 9296 and 9297 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-100% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. Substance is prepared via preparative LC/MS using the following conditions One-step purification: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20- 60% B lasted for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9296 was 11.6 mg and the purity was estimated to be 100% according to LCMS analysis. The yield of Example 9297 was 10.1 mg and the purity was estimated to be 97% according to LCMS analysis.

Example 9296

Analysis conditions A: residence time = 1.89 min; ESI-MS (+) m/z 933.25 (M+2H).

Analysis condition B: residence time = 2.56 min; ESI-MS (+) m/z 933.15 (M+2H).

Example 9297

Analysis conditions A: residence time = 1.93 min; ESI-MS (+) m/z 933.05 (M+2H).

Preparation of Examples 9298 and 9299

Examples 9298 and 9299 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19×200 Mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B was kept for 5 minutes; flow rate: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9298 was 4.1 mg and the purity was estimated to be 100% according to LCMS analysis. The yield of Example 9299 was 4.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Example 9298

Analysis conditions A: residence time = 1.81 min; ESI-MS (+) m/z 927.30 (M+2H).

Example 9299

Analysis conditions A: residence time = 1.83 min; ESI-MS (+) m/z 926.85 (M+2H).

Analysis conditions B: residence time = 3.13 min; ESI-MS (+) m/z 927.7 (M+2H).

Preparation of Examples 9300 and 9301

Examples 9300 and 9301 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9300 was 8.4 mg and the purity was estimated to be 93% according to LCMS analysis. The yield of Example 9301 was 2.1 mg and the purity was estimated to be 98% according to LCMS analysis.

Example 9300

Analysis conditions A: residence time = 1.76 min; ESI-MS (+) m/z 935.10 (M+2H).

Analysis condition B: residence time = 3.24 min; ESI-MS (+) m/z 934.85 (M+2H).

Example 9301

Analysis conditions A: residence time = 1.79 min; ESI-MS (+) m/z 935.55 (M+2H).

Analysis conditions B: residence time = 3.35 min; ESI-MS (+) m/z 935.05 (M+2H).

Preparation of Examples 9302 and 9303

Examples 9302 and 9303 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9302 was 14.2 mg and its purity was estimated to be 100% according to LCMS analysis. The yield of Example 9303 was 9.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Example 9302

Analysis conditions A: residence time = 1.71 min; ESI-MS (-) m/z 939.55 (M-2H).

Example 9303

Analysis conditions A: residence time = 1.76 min; ESI-MS (-) m/z 939.25 (M-2H).

Preparation of Examples 9304 and 9305

Examples 9304 and 9305 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9304 was 8.9 mg and its purity was estimated to be 100% according to LCMS analysis. The yield of Example 9305 was 10.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Example 9304

Analysis conditions A: residence time = 1.76 min; ESI-MS (+) m/z 934.75 (M+2H).

Example 9305

Analysis condition A: residence time = 1.78 min; ESI-MS (+) m/z (M+2H).

Preparation of Examples 9306 and 9307

Examples 9306 and 9307 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9306 was 1.3 mg and the purity was estimated to be 100% according to LCMS analysis. The yield of Example 9307 was 2.1 mg and the purity was estimated to be 96% according to LCMS analysis.

Example 9306

Analysis conditions A: residence time = 1.78 min; ESI-MS (+) m/z 940.9 (M+2H).

Analysis conditions B: residence time = 3.27 min; ESI-MS (+) m/z 941.0 (M+2H).

Example 9307

Analysis conditions A: residence time = 1.82 min; ESI-MS (+) m/z 941.5 (M+2H).

Analysis conditions B: residence time = 3.30 min; ESI-MS (+) m/z 940.9 (M+2H).

Preparation of Examples 9308 and 9309

Examples 9308 and 9309 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 150 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 40 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9308 was 2.2 mg and the purity was estimated to be 97% according to LCMS analysis. The yield of Example 9309 was 4.0 mg and the purity was estimated to be 96% according to LCMS analysis.

Example 9308

Analysis conditions A: residence time = 1.79 min; ESI-MS (+) m/z 934.3 (M+2H).

Analysis condition B: residence time = 3.25 min; ESI-MS (+) m/z 934.3 (M+2H).

Example 9309

Analysis conditions A: residence time = 1.81 min; ESI-MS (+) m/z 934.2 (M+2H).

Analysis condition B: residence time = 3.28 min ; ESI-MS (+) m/z 934.2 (M+2H).

Preparation of Examples 9310 and 9311

Examples 9310 and 9311 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 40 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9310 was 3.1 mg and the purity was estimated to be 96% according to LCMS analysis. The yield of Example 9311 was 1.4 mg and its purity was estimated to be 96% according to LCMS analysis.

Example 9310

Analysis conditions A: residence time = 1.79 min; ESI-MS (+) m/z 926.8 (M+2H).

Analysis conditions B: residence time = 3.25 min; ESI-MS (+) m/z 926.8 (M+2H).

Example 9311

Analysis conditions A: residence time = 1.83 min; ESI-MS (+) m/z 927.0 (M+2H).

Analysis condition B: residence time = 3.29 min; ESI-MS (+) m/z 926.8 (M+2H).

Preparation of Examples 9312 and 9313

Examples 9312 and 9313 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95 : 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. Consolidation containing the desired product The fraction was dissolved and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9312 was 3.3 mg and the purity was estimated to be 100% according to LCMS analysis. The yield of Example 9313 was 1.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Example 9312

Analysis conditions A: residence time = 1.29 min; ESI-MS (+) m/z 949.7 (M+2H).

Analysis condition B: residence time = 2.60 min; ESI-MS (+) m/z 949.9 (M+2H).

Example 9313

Analysis conditions A: residence time = 1.33 min; ESI-MS (+) m/z 949.8 (M+2H).

Analysis conditions B: residence time = 2.63 min; ESI-MS (+) m/z 950.0 (M+2H).

Preparation of Example 9314

Example 9314 was prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.9 mg and the purity was estimated to be 96% according to LCMS analysis.

Example 9314

Analysis conditions A: residence time = 1.35 min; ESI-MS (+) m/z 942.9 (M+2H).

Analysis condition B: residence time = 2.61 min; ESI-MS (+) m/z 942.9 (M+2H).

Preparation of Examples 9315 and 9316

Examples 9315 and 9316 were prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 20 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 9315 was 2.1 mg and the purity was estimated to be 100% according to LCMS analysis. The yield of Example 9316 was 1.3 mg and the purity was estimated to be 96% according to LCMS analysis.

Example 9315

Analysis conditions A: residence time = 1.41 min; ESI-MS (+) m/z 948.3 (M+2H).

Analysis conditions B: residence time = 2.76 min; ESI-MS (+) m/z 948.8 (M+2H).

Example 9316

Analysis conditions A: residence time = 1.46 min; ESI-MS (+) m/z 948.4 (M+2H).

Analysis conditions B: residence time = 2.81 min; ESI-MS (+) m/z 948.9 (M+2H).

Preparation of Example 9317

Example 9317 was prepared according to the method outlined in Example 10541. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 10-50% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.0 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.78 min; ESI-MS (+) m/z 942.30 (M+2H).

Analysis condition B: residence time = 2.21 min; ESI-MS (+) m/z 942.05 (M+2H).

Preparation of Example 9318

The crude material 9318 of the example was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.9 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 2.11 min; ESI-MS (+) m/z 941.45 (M+2H).

Preparation of Example 9319

The crude material of Example 9319 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.49 min; ESI-MS (+) m/z 944.7 (M+2H).

Analysis conditions B: residence time = 2.77 min; ESI-MS (+) m/z 944.2 (M+2H).

Preparation of Example 9320

The crude material of Example 9320 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.61 min; ESI-MS (+) m/z 950.4 (M+2H).

Analysis conditions B: residence time = 2.96 min; ESI-MS (+) m/z 950.6 (M+2H).

Preparation of Example 9321

The crude material of Example 9321 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 10.1 - 90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 38.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.58 min; ESI-MS (+) m/z 951.7 (M+2H).

Analysis condition B: residence time = 3.00 min; ESI-MS (+) m/z 951.8 (M+2H).

Preparation of Example 9322

The crude material of Example 9322 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.9 mg and the purity was estimated to be 94% according to LCMS analysis.

Analysis conditions A: residence time = 1.69 min; ESI-MS (+) m/z 975.0 (M+2H).

Analysis conditions B: residence time = 3.09 min; ESI-MS (+) m/z 975.1 (M+2H).

Preparation of Example 9323

The crude material of Example 9323 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.7 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.68 min; ESI-MS (+) m/z 968.2 (M+2H).

Analysis conditions B: residence time = 3.09 min; ESI-MS (+) m/z 968.4 (M+2H).

Preparation of Example 9324

The crude material of Example 9234 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.4 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.67 min; ESI-MS (+) m/z 961.2 (M+2H).

Analysis condition B: residence time = 3.08 min; ESI-MS (+) m/z 961.5 (M+2H).

Preparation of Example 9325

The crude material of Example 9325 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.5 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.64 min; ESI-MS (-) m/z 952.2 (M-2H).

Analysis conditions B: residence time = 3.02 min; ESI-MS (+) m/z 954.3 (M+2H).

Preparation of Example 9326

The crude material of Example 9226 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 35.5 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z 976.0 (M+2H).

Analysis conditions B: residence time = 2.82 min; ESI-MS (+) m/z 976.2 (M+2H).

Preparation of Example 9327

The crude material of Example 9327 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.46 min; ESI-MS (+) m/z 968.6 (M+2H).

Analysis conditions B: residence time = 2.86 min; ESI-MS (+) m/z 968.9 (M+2H).

Preparation of Example 9328

The crude material of Example 9328 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 60-90% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 34.0 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.56 min; ESI-MS (+) m/z 975.2 (M+2H).

Analysis conditions B: residence time = 2.96 min; ESI-MS (+) m/z 975.6 (M+2H).

Preparation of Example 9329

The crude material of Example 9329 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 60-90% B for 25 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.58 min; ESI-MS (+) m/z 968.0 (M+2H).

Analysis conditions B: residence time = 2.98 min; ESI-MS (+) m/z 968.2 (M+2H).

Preparation of Example 9330

The crude material of Example 9330 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.53 min; ESI-MS (+) m/z 951.1 (M+2H).

Analysis conditions B: residence time = 2.78 min; ESI-MS (+) m/z 950.7 (M+2H).

Preparation of Example 9331

The crude material of Example 9331 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.53 min; ESI-MS (+) m/z 958.5 (M+2H).

Analysis conditions B: residence time = 2.91 min; ESI-MS (+) m/z 958.3 (M+2H).

Preparation of Example 9332

The crude material of Example 9332 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.4 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.57 min; ESI-MS (+) m/z 951.6 (M+2H).

Analysis conditions B: residence time = 2.92 min; ESI-MS (+) m/z 951.0 (M+2H).

Preparation of Example 9333

The crude material of Example 9333 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.54 min; ESI-MS (+) m/z 944.2 (M+2H).

Analysis conditions B: residence time = 2.91 min; ESI-MS (+) m/z 944.3 (M+2H).

Preparation of Example 9334

The crude material of Example 9334 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 27.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.50 min; ESI-MS (+) m/z 937.5 (M+2H).

Analysis conditions B: residence time = 2.83 min; ESI-MS (+) m/z 937.4 (M+2H).

Preparation of Example 9335

The crude material of Example 9335 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19×mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 34.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.32 min; ESI-MS (+) m/z 959.3 (M+2H).

Analysis conditions B: residence time = 2.58 min; ESI-MS (+) m/z 959.0 (M+2H).

Preparation of Example 9336

The crude material of Example 9336 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19×mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.34 min; ESI-MS (+) m/z 952.0 (M+2H).

Analysis conditions B: residence time = 2.64 min; ESI-MS (+) m/z 951.7 (M+2H).

Preparation of Example 9337

The crude material of Example 9337 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19×mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 32.9 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.46 min; ESI-MS (+) m/z 959.2 (M+2H).

Analysis conditions B: residence time = 2.74 min; ESI-MS (+) m/z 958.8 (M+2H).

Preparation of Example 9338

The crude material of Example 9338 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 38.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.47 min; ESI-MS (+) m/z 951.5 (M+2H).

Analysis conditions B: residence time = 2.75 min; ESI-MS (+) m/z 951.7 (M+2H).

Preparation of Example 9345

The crude material of Example 9345 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 43.1 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions B: residence time = 3.35 min; ESI-MS (+) m/z 951.35 (M+2H).

Preparation of Example 9346

The crude material of Example 9346 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 39.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition B: residence time = 3.26 min; ESI-MS (+) m/z 944.35 (M+2H).

Preparation of Example 9344

The crude material of Example 9427 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 45.0 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis condition B: residence time = 3.26 min; ESI-MS (+) m/z 937.30 (M+2H).

Preparation of Example 9348

The crude material of Example 9348 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 39.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition B: residence time = 3.17 min; ESI-MS (+) m/z 930.35 (M+2H).

Preparation of Example 9349

The crude material of Example 9349 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 39.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.46 min; ESI-MS (+) m/z 951.70 (M+2H).

Analysis conditions B: residence time = 2.95 min; ESI-MS (+) m/z 951.65 (M+2H).

Preparation of Example 9350

The crude material of Example 9350 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 37.7 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis condition A: residence time = 1.46 min; ESI-MS (-) m/z 942.8 (M-2H).

Analysis conditions B: residence time = 2.69 min; ESI-MS (+) m/z 944.8 (M+2H).

Preparation of Example 9351

The crude material of Example 9351 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 45.3 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition A: residence time = 1.52 min; ESI-MS (-) m/z 950.0 (M-2H).

Analysis condition B: residence time = 2.70 min; ESI-MS (+) m/951.3 (M+2H).

Preparation of Example 9352

The crude material of Example 9352 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 40.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.60 min; ESI-MS (-) m/z 942.8 (M-2H).

Analysis conditions B: residence time = 2.73 min; ESI-MS (+) m/z 944.6 (M+2H).

Preparation of Example 9353

The crude material of Example 9353 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 34.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.67 min; ESI-MS (-) m/z 955.8 (M-2H).

Analysis conditions B: residence time = 2.94 min; ESI-MS (+) m/z 957.8 (M+2H).

Preparation of Example 9354

The crude material of Example 9354 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 20 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 37.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.70 min; ESI-MS (+) m/z 951.2 (M+2H).

Analysis conditions B: residence time = 2.95 min; ESI-MS (+) m/z 950.6 (M+2H).

Preparation of Example 9355

The crude material of Example 9355 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 32.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.71 min; ESI-MS (+) m/z 941.9 (M+2H).

Analysis condition B: residence time = 2.95 min; ESI-MS (-) m/z 941.6 (M-2H).

Preparation of Example 9356

The crude material of Example 9356 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.7 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis condition A: residence time = 1.58 min; ESI-MS (-) m/z 934.7 (M-2H).

Analysis conditions B: residence time = 2.82 min; ESI-MS (+) m/z 936.8 (M+2H).

Preparation of Example 9357

The crude material of Example 9257 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 49.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analytical conditions A: residence time = 1.50 min; ESI-MS (-) m/z 956.3 (M-2H).

Analysis conditions B: residence time = 2.73 min; ESI-MS (+) m/z 958.4 (M+2H).

Preparation of Example 9358

The crude material of Example 9358 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 48.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.50 min; ESI-MS (-) m/z 949.9 (M-2H).

Analysis conditions B: residence time = 2.76 min; ESI-MS (+) m/z 951.3 (M+2H).

Preparation of Example 9359

The crude material of Example 9359 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.4 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis condition A: residence time = 1.59 min; ESI-MS (-) m/z 955.7 (M-2H).

Analysis conditions B: residence time = 2.86 min; ESI-MS (+) m/z 957.8 (M+2H).

Preparation of Example 9360

The crude material of Example 9360 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 20 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 34.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.65 min; ESI-MS (-) m/z 948.9 (M-2H).

Analysis conditions B: residence time = 2.89 min; ESI-MS (+) m/z 950.8 (M+2H).

Preparation of Example 9361

The crude material of Example 9361 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 38.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.73 min; ESI-MS (-) m/z 941.2 (M-2H).

Analysis conditions B: residence time = 2.97 min; ESI-MS (+) m/z 943.4 (M+2H).

Preparation of Example 9362

The crude material of Example 9362 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 37.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.72 min; ESI-MS (+) m/z 948.6 (M+2H).

Analysis conditions B: residence time = 2.98 min; ESI-MS (+) m/z 950.9 (M+2H).

Preparation of Example 9363

The crude material of Example 9363 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.68 min; ESI-MS (-) m/z 941.5 (M-2H).

Analysis conditions B: residence time = 2.98 min; ESI-MS (+) m/z 943.4 (M+2H).

Preparation of Example 9364

The crude material of Example 9364 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 42.7 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.71 min; ESI-MS (+) m/z 934.7 (M+2H).

Analysis conditions B: residence time = 2.97 min; ESI-MS (+) m/z 936.8 (M+2H).

Preparation of Example 9365

The crude material of Example 9356 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 48-88% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.5 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition A: residence time = 1.67 min; ESI-MS (-) m/z 927.8 (M-2H).

Analysis conditions B: residence time = 2.93 min; ESI-MS (+) m/z 929.6 (M+2H).

Preparation of Example 9366

The crude material of Example 9366 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 39.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition A: residence time = 1.56 min; ESI-MS (-) m/z 949.2 (M-2H).

Analysis conditions B: residence time = 2.78 min; ESI-MS (+) m/z 951.2 (M+2H).

Preparation of Example 9376

The crude material of Example 9367 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition A: residence time = 1.77 min; ESI-MS (-) m/z 965.8 (M-2H).

Analysis conditions B: residence time = 2.98 min; ESI-MS (+) m/z 967.5 (M+2H).

Preparation of Example 9368

The crude material of Example 9368 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 38.8 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.65 min; ESI-MS (+) m/z 968.7 (M+2H).

Analysis conditions B: residence time = 2.90 min; ESI-MS (+) m/z 968.4 (M+2H).

Preparation of Example 9369

The crude material of Example 9369 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.3 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.62 min; ESI-MS (+) m/z 948.6 (M+2H).

Analysis conditions B: residence time = 2.78 min; ESI-MS (+) m/z 950.8 (M+2H).

Preparation of Example 9370

The crude material of Example 9370 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 47.0 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.58 min; ESI-MS (+) m/z 952.1 (M+2H).

Analysis conditions B: residence time = 2.83 min; ESI-MS (+) m/z 951.2 (M+2H).

Preparation of Example 9371

The crude material of Example 9371 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.3 mg and the purity was estimated to be 94% according to LCMS analysis.

Analysis condition A: residence time = 1.69 min; ESI-MS (-) m/z 966.0 (M-2H).

Analysis conditions B: residence time = 2.85 min; ESI-MS (+) m/z 968.2 (M+2H).

Preparation of Example 9372

The crude material of Example 9372 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.77 min; ESI-MS (-) m/z 966.1 (M-2H).

Analysis conditions B: residence time = 2.95 min; ESI-MS (+) m/z 968.3 (M+2H).

Preparation of Example 9373

The crude material of Example 9173 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.56 min; ESI-MS (+) m/z 950.6 (M+2H).

Analysis conditions B: residence time = 2.68 min; ESI-MS (+) m/z 950.5 (M+2H).

Preparation of Example 9374

The crude material of Example 9374 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.67 min; ESI-MS (-) m/z 949.3 (M-2H).

Analysis conditions B: residence time = 2.82 min; ESI-MS (+) m/z 951.2 (M+2H).

10000 series

Examples 10012 through 10043, examples 10045 through 10052, examples 10122 through 10147, and examples 10196 through 10219 were prepared following "general synthetic sequence A."

Example 10044, Examples 10053 through 10121, and Examples 10148 through 10195 were prepared following "General Synthesis Sequence D."

Preparation of Example 10012

The crude material of Example 10012 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.9 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.54 min; ESI-MS (+) m/z 954.0 (M+2H).

Analysis conditions B: residence time = 2.72 min; ESI-MS (+) m/z 953.5 (M+2H).

ESI-HRMS (+) m/z: Calculated: 149.

Preparation of Example 10013

The crude material of Example 10013 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.2 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.46 min; ESI-MS (+) m/z 936.9 (M+2H).

Analysis condition B: residence time = 2.58 min; ESI-MS (+) m/z 937.5 (M+2H).

ESI-HRMS (+) m/z: Calculated: </RTI> </ RTI> </ RTI> </ RTI> </ RTI> <RTIgt;

Preparation of Example 10014

The crude material of Example 10014 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.44 min; ESI-MS (+) m/z 945.3 (M+2H).

Analysis conditions B: residence time = 2.56 min; ESI-MS (+) m/z 945.1 (M+2H).

ESI-HRMS (+) m/z: Calculated: 944.4 </RTI> (M+2H);

Preparation of Example 10015

The crude material of Example 10015 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.2 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.61 min; ESI-MS (+) m/z 918.5 (M+2H).

Analysis conditions B: residence time = 2.79 min; ESI-MS (+) m/z 918.5 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10016

The crude material of Example 10016 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.6 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition A: residence time = 1.61 min; ESI-MS (-) m/z 924.4 (M-2H).

Analysis conditions B: residence time = 2.77 min; ESI-MS (+) m/z 926.6 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10017

The crude material of Example 10017 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.53 min; ESI-MS (+) m/z 910.4 (M+2H).

Analysis conditions B: residence time = 2.64 min; ESI-MS (+) m/z 910.0 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10018

The crude material of Example 10018 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.5 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.45 min; ESI-MS (+) m/z 918.9 (M+2H).

Analysis conditions B: residence time = 2.61 min; ESI-MS (+) m/z 918.0 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10019

The crude material of Example 10019 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.3 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis condition A: residence time = 1.64 min; ESI-MS (+) m/z 1864.6 (M+H).

Analysis condition B: residence time = 2.79 min; ESI-MS (+) m/z 933.0 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10020

The crude material of Example 10020 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.2 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition A: retention time = 1.68min; ESI-MS (+ ) m / z 940.8 (M + 2H).

Analysis conditions B: residence time = 2.81 min; ESI-MS (+) m/z 940.6 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Examples 10021 and 10022

The crude material of Example 10021 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.2 mg and the purity was estimated to be 90% according to LCMS analysis.

Analysis condition A: residence time = 1.52 min; ESI-MS (-) m/z 922.0 (M-2H).

Analysis conditions B: residence time = 2.64 min; ESI-MS (+) m/z 924.1 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10022

The crude material of Example 10022 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.8 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition A: residence time = 1.61 min; ESI-MS (-) m/z 924.0 (M-2H).

Analysis conditions B: residence time = 2.77 min; ESI-MS (+) m/z 925.9 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10023

The crude material of Example 10023 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.61 min; ESI-MS (-) m/z 932.1 (M-2H).

Analysis conditions B: residence time = 2.76 min; ESI-MS (+) m/z 934.1 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10024

The crude material of Example 10024 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.46 min; ESI-MS (+) m/z 925.1 (M+2H).

Analysis conditions B: residence time = 2.60 min; ESI-MS (+) m/z 925.4 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10025

The crude material of Example 10025 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.66 min; ESI-MS (-) m/z 923.6 (M-2H).

Analysis conditions B: residence time = 2.82 min; ESI-MS (+) m/z 925.6 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10034

The crude material of Example 10034 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.5 mg and the purity was estimated to be 90% according to LCMS analysis.

Analysis conditions A: residence time = 1.61 min; ESI-MS (+) m/z 916.60 (M+2H).

Analysis conditions B: residence time = 2.19 min; ESI-MS (+) m/z 916.55 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10035

The crude material of Example 10035 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.68 min; ESI-MS (-) m/z 931.6 (M-2H).

Analysis conditions B: residence time = 3.10 min; ESI-MS (+) m/z 934.10 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10036

The crude material of Example 10036 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.1 mg and the purity was estimated to be 91% according to LCMS analysis.

Analysis conditions A: residence time = 1.78 min; ESI-MS (+) m/z 916.80 (M+2H).

Analysis condition B: residence time = 2.61 min; ESI-MS (+) m/z 916.80 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10037

The crude material of Example 10037 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.63 min; ESI-MS (+) m/z 924.55 (M+2H).

Analysis condition B: residence time = 2.95 min; ESI-MS (+) m/z 924.65 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10042

The crude material of Example 10042 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.68 min; ESI-MS (+) m/z 945.40 (M+2H).

Analysis condition B: residence time = 3.13 min; ESI-MS (+) m/z 945.40 (M+2H).

ESI-HRMS (+) m/z: Calculated: 149.

Preparation of Example 10043

The crude material of Example 10043 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.78 min; ESI-MS (+) m/z 931.80 (M+2H).

Analysis conditions B: residence time = 2.57 min; ESI-MS (+) m/z 932.00 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10044

The crude material of Example 10044 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.4 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.68 min; ESI-MS (+) m/z 897.95 (M+2H).

Analysis conditions B: residence time = 3.31 min; ESI-MS (+) m/z 897.85 (M+2H).

Preparation of Example 10045

The crude material of Example 10045 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.5 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.70 min; ESI-MS (+) m/z 956.3 (M+2H).

Analysis conditions B: residence time = 3.18 min; ESI-MS (+) m/z 956.3 (M+2H).

ESI-HRMS (+) m/z: Calculated: 955.9 s (M+2H);

Preparation of Example 10047

The crude material of Example 10047 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 38.6 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.71 min; ESI-MS (+) m/z 957.35 (M+2H).

Analysis condition B: residence time = 3.17 min; ESI-MS (+) m/z 957.30 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10049

The crude material of Example 10049 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 36.2 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.69 min; ESI-MS (+) m/z 932.45 (M+2H).

Analysis condition B: residence time = 3.14 min; ESI-MS (+) m/z 932.45 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10050

The crude material of Example 10050 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.0 mg and the purity was estimated to be 99% according to LCMS analysis.

Analytical conditions A: residence time = 1.70 min; ESI-MS (+) m/z 940.40 (M+2H).

Analysis conditions B: residence time = 3.14 min; ESI-MS (+) m/z 940.40 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 10051

The crude material of Example 10051 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.1 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.57 min; ESI-MS (+) m/z 923.90 (M+2H).

Analysis conditions B: residence time = 2.95 min; ESI-MS (+) m/z 923.90 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10052

The crude material of Example 10052 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.54 min; ESI-MS (+) m/z 931.90 (M+2H).

Analysis conditions B: residence time = 2.94 min; ESI-MS (+) m/z 931.90 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10053

The crude material of Example 10053 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 36.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.79 min; ESI-MS (+) m/z 924.90 (M+2H).

Analysis conditions B: residence time = 3.37 min; ESI-MS (+) m/z 925.00 (M+2H).

Preparation of Example 10054

The crude material of Example 10054 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 32.5 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.89 min; ESI-MS (+) m/z 883.95 (M+2H).

Analysis conditions B: residence time = 3.38 min; ESI-MS (+) m/z 883.85 (M+2H).

Preparation of Example 10055

The crude material of Example 10055 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.96 min; ESI-MS (+) m/z 892.15 (M+2H).

Analysis condition B: residence time = 3.09 min; ESI-MS (+) m/z 892.20 (M+2H).

Preparation of Example 10056

The crude material of Example 10056 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 41.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.91 min; ESI-MS (+) m/z 903.50 (M+2H).

Analysis condition B: residence time = 3.40 min; ESI-MS (+) m/z 903.65 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10057

The crude material of Example 10057 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.79 min; ESI-MS (+) m/z 890.70 (M+2H).

Analysis condition B: residence time = 3.26 min; ESI-MS (+) m/z 891.05 (M+2H).

Preparation of Example 10058

The crude material of Example 10058 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.92 min; ESI-MS (+) m/z 890.60 (M+2H).

Analysis condition B: residence time = 3.43 min; ESI-MS (+) m/z 890.65 (M+2H).

Preparation of Example 10059

The crude material of Example 10059 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 2.03 min; ESI-MS (+) m/z 899.00 (M+2H).

Analysis condition B: residence time = 3.15 min; ESI-MS (+) m/z 899.10 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10060

The crude material of Example 10060 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 27.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.97 min; ESI-MS (+) m/z 910.60 (M+2H).

Analysis condition B: residence time = 3.46 min; ESI-MS (+) m/z 910.60 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10061

The crude material of Example 10061 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.3 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.84 min; ESI-MS (+) m/z 897.60 (M+2H).

Analysis conditions B: residence time = 3.33 min; ESI-MS (+) m/z 897.60 (M+2H).

Preparation of Example 10062

The crude material of Example 10062 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.95 min; ESI-MS (+) m/z 897.30 (M+2H).

Analysis conditions B: residence time = 3.49 min; ESI-MS (+) m/z 897.75 (M+2H).

Preparation of Example 10063

The crude material of Example 10063 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 2.07 min; ESI-MS (+) m/z 906.05 (M+2H).

Analysis conditions B: residence time = 3.16 min; ESI-MS (+) m/z 906.20 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10064

The crude material of Example 10064 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 2.01 min; ESI-MS (+) m/z 917.85 (M+2H).

Analysis conditions B: residence time = 3.05 min; ESI-MS (+) m/z 917.70 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10065

The crude material of Example 10065 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.89 min; ESI-MS (+) m/z 904.75 (M+2H).

Analysis conditions B: residence time = 3.36 min; ESI-MS (+) m/z 904.45 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10066

The crude material of Example 10066 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.4 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 2.02 min; ESI-MS (+) m/z 937.65 (M+2H).

Analysis conditions B: residence time = 3.07 min; ESI-MS (+) m/z 937.65 (M+2H).

Preparation of Example 10067

The crude material of Example 10067 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.95 min; ESI-MS (+) m/z 910.55 (M+2H).

Analysis condition B: residence time = 3.43 min; ESI-MS (+) m/z 910.60 (M+2H).

Preparation of Example 10068

The crude material of Example 10068 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.83 min; ESI-MS (+) m/z 933.60 (M+2H).

Analysis condition B: residence time = 2.53 min; ESI-MS (+) m/z 933.60 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10069

The crude material of Example 10069 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 2.02 min; ESI-MS (+) m/z 933.20 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10070

The crude material of Example 10070 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.0 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 2.05 min; ESI-MS (+) m/z 920.00 (M+2H).

Analysis conditions B: residence time = 2.79 min; ESI-MS (+) m/z 919.60 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10071

The crude material of Example 10071 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.94 min; ESI-MS (+) m/z 918.75 (M+2H).

Analysis conditions B: residence time = 3.46 min; ESI-MS (+) m/z 919.75 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10072

The crude material of Example 10072 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.5 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.80 min; ESI-MS (+) m/z 933.55 (M+2H).

Analysis condition B: residence time = 2.48 min; ESI-MS (+) m/z 933.00 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10073

The crude material of Example 10073 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.97 min; ESI-MS (+) m/z 933.45 (M+2H).

Analysis conditions B: residence time = 2.67 min; ESI-MS (+) m/z 932.90 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10074

The crude material of Example 10074 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 2.01 min; ESI-MS (+) m/z 919.60 (M+2H).

ESI-HRMS (+) m/z: Calculated:

Preparation of Example 10075

The crude material of Example 10075 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.90 min; ESI-MS (+) m/z 919.10 (M+2H).

Analysis conditions B: residence time = 2.62 min; ESI-MS (+) m/z 919.00 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10076

The crude material of Example 10076 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.75 min; ESI-MS (+) m/z 910.85 (M+2H).

Analysis conditions B: residence time = 2.36 min; ESI-MS (+) m/z 910.60 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10077

The crude material of Example 10077 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.3 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 2.44 min; ESI-MS (+) m/z 924.65 (M+2H).

Analysis condition B: residence time = 3.37 min; ESI-MS (+) m/z 924.70 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10078

The crude material of Example 10078 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.6 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.84 min; ESI-MS (+) m/z 929.05 (M+2H).

Analysis condition B: residence time = 3.46 min; ESI-MS (+) m/z 929.20 (M+2H).

ESI-HRMS (+) m/z: Calculated: </ RTI> </ RTI> </ RTI> </ RTI> <RTIgt;

Preparation of Example 10079

The crude material of Example 10079 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.1 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.65 min; ESI-MS (+) m/z 926.55 (M+2H).

Analysis conditions B: residence time = 3.18 min; ESI-MS (+) m/z 926.60 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10080

The crude material of Example 10080 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 0.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.59 min; ESI-MS (+) m/z 897.70 (M+2H).

Analysis condition B: residence time = 3.14 min; ESI-MS (+) m/z 897.70 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10081

The crude material of Example 10081 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.3 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.72 min; ESI-MS (+) m/z 904.60 (M+2H).

Analysis conditions B: residence time = 2.32 min; ESI-MS (+) m/z 904.55 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10082

The crude material of Example 10082 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.73 min; ESI-MS (+) m/z 906.70 (M+2H).

Analysis conditions B: residence time = 3.29 min; ESI-MS (+) m/z 906.65 (M+2H).

ESI-HRMS (+) m/z: Calculated: </RTI> <RTIgt; </RTI> <RTIgt;

Preparation of Example 10083

The crude material of Example 10083 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z 905.80 (M+2H).

Analysis conditions B: residence time = 2.90 min; ESI-MS (+) m/z 905.70 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10084

The crude material of Example 10084 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.6 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.81 min; ESI-MS (+) m/z 904.45 (M+2H).

Analysis condition B: residence time = 3.36 min; ESI-MS (+) m/z 904.60 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10085

The crude material of Example 10085 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.77 min; ESI-MS (+) m/z 907.35 (M+2H).

Analysis conditions B: residence time = 3.38 min; ESI-MS (+) m/z 906.50 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10086

The crude material of Example 10086 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.63 min; ESI-MS (+) m/z 905.65 (M+2H).

Analysis condition B: residence time = 2.98 min; ESI-MS (+) m/z 905.65 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10087

The crude material of Example 10087 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.80 min; ESI-MS (+) m/z 917.45 (M+2H).

Analysis conditions B: residence time = 3.40 min; ESI-MS (+) m/z 917.70 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10088

The crude material of Example 10088 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.72 min; ESI-MS (+) m/z 919.90 (M+2H).

Analysis condition B: residence time = 3.33 min; ESI-MS (+) m/z 919.95 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10089

The crude material of Example 10089 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition A: residence time = 2.14 min; ESI-MS (-) m/z 945.30 (M-2H).

Analysis condition B: residence time = 3.19 min; ESI-MS (+) m/z 947.10 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10090

The crude material of Example 10090 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.6 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis condition B: retention time = 2.98min; ESI-MS (+ ) m / z 944.20 (M + 2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10091

The crude material of Example 10091 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.7 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis condition A: residence time = 1.75 min; ESI-MS (-) m/z 935.2 (M-2H).

Analysis conditions B: residence time = 3.01 min; ESI-MS (+) m/z 936.7 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 10092

The crude material of Example 10092 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.5 mg and the purity was estimated to be 94% according to LCMS analysis.

Analysis conditions B: residence time = 3.14 min; ESI-MS (+) m/z 961.2 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10093

The crude material of Example 10093 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.5 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition A: retention time = 1.82min; ESI-MS (- ) m / z 959.0 (M-2H).

Analysis conditions B: residence time = 3.09 min; ESI-MS (+) m/z 960.9 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10094

The crude material of Example 10094 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.4 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.93 min; ESI-MS (+) m/z 938.70 (M+2H).

Analysis condition B: residence time = 3.09 min; ESI-MS (+) m/z 939.30 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10095

The crude material of Example 10095 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.75 min; ESI-MS (+) m/z 946.30 (M+2H).

Analysis conditions B: residence time = 3.08 min; ESI-MS (+) m/z 946.60 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10096

The crude material of Example 10096 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 32.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.82 min; ESI-MS (+) m/z 959.10 (M+2H).

Analysis condition B: residence time = 3.13 min; ESI-MS (+) m/z 959.30 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10097

The crude material of Example 10097 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 44.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.60 min; ESI-MS (+) m/z 959.70 (M+2H).

Analysis condition B: residence time = 3.11 min; ESI-MS (+) m/z 959.70 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10098

The crude material of Example 10098 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 36.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.91 min; ESI-MS (+) m/z 952.10 (M+2H).

Analysis condition B: residence time = 3.30 min; ESI-MS (+) m/z 952.20 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10099

The crude material of Example 10099 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 36.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.65 min; ESI-MS (+) m/z 952.80 (M+2H).

Analysis conditions B: residence time = 2.98 min; ESI-MS (+) m/z 953.20 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10100

The crude material of Example 10100 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.5 mg and the purity was estimated to be 94% according to LCMS analysis.

Analysis conditions A: residence time = 1.83 min; ESI-MS (+) m/z 959.20 (M+2H).

Analysis condition B: residence time = 3.34 min; ESI-MS (+) m/z 959.25 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 10102

The crude material of Example 10102 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 36.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.90 min; ESI-MS (+) m/z 909.05 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10103

The crude material of Example 10103 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 27.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.88 min; ESI-MS (+) m/z 900.35 (M+2H).

Analysis condition B: retention time = 2.57min; ESI-MS (+ ) m / z 900.10 (M + 2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 10104

The crude material of Example 10104 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 40.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.94 min; ESI-MS (+) m/z 927.45 (M+2H).

Analysis condition B: residence time = 3.44 min; ESI-MS (+) m/z 927.40 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10105

The crude material of Example 10105 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 39.2 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.88 min; ESI-MS (+) m/z 901.95 (M+2H).

Analysis conditions B: residence time = 3.44 min; ESI-MS (+) m/z 902.15 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10106

The crude material of Example 10106 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 40.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.61 min; ESI-MS (+) m/z 910.20 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10107

The crude material of Example 10107 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 42.7 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.68 min; ESI-MS (+) m/z 917.50 (M+2H).

Analysis condition B: retention time = 2.75min; ESI-MS (+ ) m / z 917.10 (M + 2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10108

The crude material of Example 10108 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 31.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.84 min; ESI-MS (+) m/z 918.90 (M+2H).

Analysis condition B: residence time = 3.06 min; ESI-MS (+) m/z 918.40 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10109

The crude material of Example 10109 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 45.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.86 min; ESI-MS (+) m/z 912.40 (M+2H).

Analysis conditions B: residence time = 3.07 min; ESI-MS (+) m/z 911.50 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10110

The crude material of Example 10110 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.58 min; ESI-MS (+) m/z 919.60 (M+2H).

Analysis conditions B: residence time = 2.73 min; ESI-MS (+) m/z 919.50 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10111

The crude material of Example 10111 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.68 min; ESI-MS (+) m/z 925.95 (M+2H).

Analysis conditions B: residence time = 3.21 min; ESI-MS (+) m/z 926.00 (M+2H).

ESI-HRMS (+) m/z: Calculated: s.

Preparation of Example 10112

The crude material of Example 10112 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 41.6 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.89 min; ESI-MS (+) m/z 916.50 (M+2H).

Analysis condition B: residence time = 3.11 min; ESI-MS (+) m/z 916.50 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10113

The crude material of Example 10113 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 31.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.96 min; ESI-MS (+) m/z 909.40 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10114

The crude material of Example 10114 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 39.5 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.63 min; ESI-MS (+) m/z 916.8 (M+2H).

Analysis conditions B: residence time = 2.78 min; ESI-MS (+) m/z 917.0 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10115

The crude material of Example 10115 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 31.5 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.72 min; ESI-MS (+) m/z 923.95 (M+2H).

Analysis condition B: residence time = 3.26 min; ESI-MS (+) m/z 923.95 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10116

The crude material of Example 10116 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 35.8 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.67 min; ESI-MS (+) m/z 951.90 (M+2H).

Analysis conditions B: residence time = 3.18 min; ESI-MS (+) m/z 951.90 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10117

The crude material of Example 10117 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 32.9 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.62 min; ESI-MS (+) m/z 950.70 (M+2H).

Analysis conditions B: residence time = 2.80 min; ESI-MS (+) m/z 950.10 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 10118

The crude material of Example 10118 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.8 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.84 min; ESI-MS (+) m/z 944.45 (M+2H).

Analysis condition B: residence time = 3.34 min; ESI-MS (+) m/z 944.35 (M+2H).

ESI-HRMS (+) m/z: Calculated: 149.

Preparation of Example 10119

The crude material of Example 10119 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 32.8 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.89 min; ESI-MS (+) m/z 942.45 (M+2H).

Analysis condition B: residence time = 3.41 min; ESI-MS (+) m/z 942.35 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10120

The crude material of Example 10120 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 47.5 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.72 min; ESI-MS (+) m/z 944.90 (M+2H).

Analysis condition B: residence time = 3.41 min; ESI-MS (+) m/z 944.85 (M+2H).

ESI-HRMS (+) m/z: Calculated: 944.4311 (M+2H);

Preparation of Example 10121

The crude material of Example 10121 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 48.0 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.79 min; ESI-MS (+) m/z 942.65 (M+2H).

Analysis conditions B: residence time = 2.47 min; ESI-MS (+) m/z 942.80 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10122

The crude material of Example 10122 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.9 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.67 min; ESI-MS (+) m/z 949.80 (M+2H).

Analysis conditions B: residence time = 2.73 min; ESI-MS (+) m/z 949.60 (M+2H).

ESI-HRMS (+) m/z: Calculated: </RTI> <RTI ID=0.0></RTI> </RTI> <RTIgt;

Preparation of Example 10123

The crude material of Example 10123 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.5 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis condition A: retention time = 1.61min; ESI-MS (+ ) m / z 957.15 (M + 2H).

Analysis conditions B: residence time = 2.27 min; ESI-MS (+) m/z 957.30 (M+2H).

Preparation of Example 10126

The crude material of Example 10126 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.46 min; ESI-MS (+) m/z 956.60 (M+2H).

Analysis conditions B: residence time = 2.50 min; ESI-MS (+) m/z 956.60 (M+2H).

Preparation of Example 10127

The crude material of Example 10127 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.45 min; ESI-MS (+) m/z 942.95 (M+2H).

Analysis conditions B: residence time = 2.89 min; ESI-MS (+) m/z 942.80 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10128

The crude material of Example 10128 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.45 min; ESI-MS (+) m/z 943.45 (M+2H).

Analysis condition B: residence time = 2.84 min; ESI-MS (+) m/z 943.10 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10129

The crude material of Example 10129 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.0 mg and the purity was estimated to be 92% according to LCMS analysis.

Analysis conditions A: residence time = 1.51 min; ESI-MS (+) m/z 935.65 (M+2H).

Analysis condition B: residence time = 3.04 min; ESI-MS (+) m/z 935.60 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10130

The crude material of Example 10130 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.2 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.49 min; ESI-MS (+) m/z 935.80 (M+2H).

Analysis conditions B: residence time = 2.96 min; ESI-MS (+) m/z 936.15 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10131

The crude material of Example 10131 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.7 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.53 min; ESI-MS (+) m/z 935.10 (M+2H).

Analysis conditions B: residence time = 2.94 min; ESI-MS (+) m/z 936.20 (M+2H).

ESI-HRMS (+) m/z: Calculated: </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> <RTIgt;

Preparation of Example 10132

The crude material of Example 10132 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.49 min; ESI-MS (+) m/z 935.55 (M+2H).

Analysis condition B: residence time = 2.88 min; ESI-MS (+) m/z 935.65 (M+2H).

ESI-HRMS (+) m/z: Calculated: </ RTI> </ RTI> </ RTI> <RTIgt;

Preparation of Example 10136

The crude material of Example 10136 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.8 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.27 min; ESI-MS (+) m/z 951.00 (M+2H).

Analysis conditions B: residence time = 2.64 min; ESI-MS (+) m/z 951.20 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10137

The crude material of Example 10137 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.3 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.42 min; ESI-MS (+) m/z 928.30 (M+2H).

Analysis conditions B: residence time = 2.52 min; ESI-MS (+) m/z 928.00 (M+2H).

ESI-HRMS (+) m/z: calc.: 927.

Preparation of Example 10138

The crude material of Example 10138 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.39 min; ESI-MS (+) m/z 929.40 (M+2H).

Analysis conditions B: residence time = 2.83 min; ESI-MS (+) m/z 929.75 (M+2H).

Preparation of Example 10139

The crude material of Example 10139 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.4 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition A: retention time = 1.37min; ESI-MS (+ ) m / z 929.60 (M + 2H).

Analysis conditions B: residence time = 1.96 min; ESI-MS (+) m/z 929.70 (M+2H).

Preparation of Example 10140

The crude material of Example 10140 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z 936.20 (M+2H).

Analysis conditions B: residence time = 2.13 min; ESI-MS (+) m/z 936.20 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10141

The crude material of Example 10141 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 27.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.36 min; ESI-MS (+) m/z 950.60 (M+2H).

Analysis condition B: residence time = 1.79 min; ESI-MS (+) m/z 950.60 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10142

The crude material of Example 10142 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition B: residence time = 1.51 min; ESI-MS (+) m/z 951.90 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10143

The crude material of Example 10143 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.1 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.45 min; ESI-MS (+) m/z 928.40 (M+2H).

Analysis conditions B: residence time = 2.54 min; ESI-MS (+) m/z 928.30 (M+2H).

ESI-HRMS (+) m/z: calc.: 927.

Preparation of Example 10144

The crude material of Example 10144 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.36 min; ESI-MS (+) m/z 929.9 (M+2H).

Analysis condition B: residence time = 2.41 min; ESI-MS (+) m/z 930.5 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10145

The crude material of Example 10145 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.4 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.41 min; ESI-MS (+) m/z 929.55 (M+2H).

Analysis condition B: retention time = 1.99min; ESI-MS (+ ) m / z 930.65 (M + 2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10146

The crude material of Example 10146 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.42 min; ESI-MS (+) m/z 936.8 (M+2H).

Analysis condition B: residence time = 2.53 min; ESI-MS (+) m/z 936.7 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10147

The crude material of Example 10147 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.4 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.23 min; ESI-MS (+) m/z 951.2 (M+2H).

Analysis conditions B: residence time = 2.33 min; ESI-MS (+) m/z 951.6 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10148

The crude material of Example 10148 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.7 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition B: residence time = 1.71 min; ESI-MS (-) m/z 950.9 (M-2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10149

The crude material of Example 10149 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.2 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis condition A: residence time = 1.73 min; ESI-MS (-) m/z 943.5 (M-2H).

Analysis conditions B: residence time = 2.89 min; ESI-MS (+) m/z 945.4 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10150

The crude material of Example 10150 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.7 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis condition A: residence time = 1.61 min; ESI-MS (-) m/z 944.2 (M-2H).

Analysis conditions B: residence time = 2.84 min; ESI-MS (+) m/z 946.4 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10151

The crude material of Example 10151 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.59 min; ESI-MS (-) m/z 937.3 (M-2H).

Analysis conditions B: residence time = 2.83 min; ESI-MS (+) m/z 939.8 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10152

The crude material of Example 10152 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.72 min; ESI-MS (-) m/z 928.1 (M-2H).

Analysis condition B: residence time = 3.06 min; ESI-MS (+) m/z 930.0 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10153

The crude material of Example 10153 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.68 min; ESI-MS (-) m/z 952.6 (M-2H).

Analysis conditions B: residence time = 2.95 min; ESI-MS (+) m/z 954.8 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10154

The crude material of Example 10154 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 36.0 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z 936.7 (M+2H).

Analysis conditions B: residence time = 2.74 min; ESI-MS (+) m/z 937.5 (M+2H).

ESI-HRMS (+) m/z: Found: </RTI> </RTI>

Preparation of Example 10155

The crude material of Example 10155 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 31.4 mg and the purity was estimated to be 94% according to LCMS analysis.

Analysis condition A: residence time = 1.54 min; ESI-MS (-) m/z 928.6 (M-2H).

Analysis conditions B: residence time = 2.69 min; ESI-MS (+) m/z 930.3 (M+2H).

ESI-HRMS (+) m/z: Calculated: </RTI> </RTI>

Preparation of (R)-4-(((9H-茀-9-yl)methoxy)carbonyl)morpholine-3-carboxylic acid

Add sodium bicarbonate (1.922 g, 22.88 mmol) and FMOC-OSU (5.14) to a solution of (R)-morpholine-3-carboxylic acid (2 g, 15.25 mmol) in THF (50.8 mL) and water (25.4 mL) g, 15.25 mmol). The resulting mixture was stirred for 28 hours. After removal of the THF, the white suspension was diluted with saturated NaHCO3 / water and diethyl ether. Filter through diatomaceous earth. Wash with water and ether. The ether and aqueous layers were separated. The aqueous layer was acidified with 1N HCl, and extracted with ethyl acetate, washed with brine, dried over Na ₂ SO _4, and concentrated to give 3.19g of a white solid (R) -4 - ((( 9H- fluorene-9-yl ) methoxy)carbonyl)morpholine-3-carboxylic acid.

Preparation of Example 10156

The crude material of Example 10156 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.1 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.60 min; ESI-MS (-) m/z 943.3 (M-2H).

Analysis conditions B: residence time = 2.83 min; ESI-MS (+) m/z 945.8 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10157

The crude material of Example 10157 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.1 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis condition A: residence time = 1.59 min; ESI-MS (-) m/z 936.6 (M-2H).

Analysis condition B: residence time = 2.81 min; ESI-MS (+) m/z 938.7 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10158

The crude material of Example 10158 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.1 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.77 min; ESI-MS (+) m/z 937.4 (M+2H).

Analysis conditions B: residence time = 2.99 min; ESI-MS (+) m/z 937.4 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10159

The crude material of Example 10159 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.73 min; ESI-MS (+) m/z 960.4 (M+2H).

Analysis conditions B: residence time = 3.01 min; ESI-MS (+) m/z 962.5 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10160

The crude material of Example 10161 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.6 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.80 min; ESI-MS (+) m/z 968.80 (M+2H).

Analysis condition B: residence time = 3.44 min; ESI-MS (+) m/z 968.80 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 10161

The crude material of Example 10161 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.4 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.65 min; ESI-MS (+) m/z 944.65 (M+2H).

Analysis conditions B: residence time = 3.20 min; ESI-MS (+) m/z 944.80 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10162

The crude material of Example 10162 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 33.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.85 min; ESI-MS (+) m/z 944.45 (M+2H).

ESI-HRMS (+) m/z: Calculated: 149.

Preparation of Example 10163

The crude material of Example 10163 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.59 min; ESI-MS (+) m/z 950.30 (M+2H).

Analysis condition B: residence time = 2.90 min; ESI-MS (+) m/z 950.70 (M+2H).

ESI-HRMS (+) m/z: Calculated: 149.

Preparation of Example 10164

The crude material of Example 10164 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.6 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.75 min; ESI-MS (+) m/z 958.10 (M+2H).

Analysis conditions B: residence time = 2.98 min; ESI-MS (+) m/z 958.00 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10165

The crude material of Example 10165 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.2 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.63 min; ESI-MS (+) m/z 945.40 (M+2H).

Analysis condition B: retention time = 2.85min; ESI-MS (+ ) m / z 944.70 (M + 2H).

ESI-HRMS (+) m/z: Calculated: </ RTI> </ RTI> </ RTI> </ RTI> <RTIgt;

Preparation of Example 10166

The crude material of Example 10166 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.73 min; ESI-MS (+) m/z 951.50 (M+2H).

Analysis conditions B: residence time = 2.98 min; ESI-MS (+) m/z 951.00 (M+2H).

Preparation of Example 10167

The crude material of Example 10167 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 48.2 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.61 min; ESI-MS (+) m/z 949.50 (M+2H).

Analysis condition B: retention time = 2.77min; ESI-MS (+ ) m / z 949.50 (M + 2H).

Preparation of Example 10168

The crude material of Example 10168 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.74 min; ESI-MS (+) m/z 967.80 (M+2H).

Analysis condition B: residence time = 3.10 min; ESI-MS (+) m/z 968.10 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 10169

The crude material of Example 10169 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.9 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis condition A: retention time = 1.75min; ESI-MS (+ ) m / z 937.10 (M + 2H).

Analysis condition B: residence time = 3.17 min; ESI-MS (+) m/z 937.30 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10170

The crude material of Example 10170 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.70 min; ESI-MS (+) m/z 919.20 (M+2H).

Analysis conditions B: residence time = 2.39 min; ESI-MS (+) m/z 920.05 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10171

The crude material of Example 10171 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 35.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.59 min; ESI-MS (+) m/z 935.50 (M+2H).

Analysis conditions B: residence time = 2.93 min; ESI-MS (+) m/z 935.50 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10172

The crude material of Example 10172 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 32.9 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition A: residence time = 1.69 min; ESI-MS (-) m/z 927.65 (M-2H).

Analysis conditions B: residence time = 2.35 min; ESI-MS (+) m/z 929.65 (M+2H).

ESI-HRMS (+) m/z: Calculated: </RTI> </RTI>

Preparation of Example 10173

The crude material of Example 10173 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.6 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: retention time = 1.45min; ESI-MS (+ ) m / z 938.0 (M + 2H).

Analysis condition B: retention time = 2.70min; ESI-MS (+ ) m / z 938.3 (M + 2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10174

The crude material of Example 10174 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.7 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.58 min; ESI-MS (+) m/z 944.1 (M+2H).

Analysis conditions B: residence time = 2.95 min; ESI-MS (+) m/z 943.5 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10175

The crude material of Example 10175 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 38.7 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis condition A: residence time = 1.89 min; ESI-MS (-) m/z 918.8 (M-2H).

Analysis condition B: residence time = 3.34 min; ESI-MS (+) m/z 921.15 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10176

The crude material of Example 10176 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.1 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis condition A: residence time = 1.37 min; ESI-MS (-) m/z 973.9 (M-2H).

Analysis conditions B: residence time = 2.57 min; ESI-MS (+) m/z 976.1 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10177

The crude material of Example 10177 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.6 mg and the purity was estimated to be 96% according to LCMS analysis.

Analytical conditions A: residence time = 1.55 min ; ESI-MS (+) m/z 959.5 (M+2H).

Analysis conditions B: residence time = 2.95 min; ESI-MS (+) m/z 959.2 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10178

The crude material of Example 10178 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B : 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 46.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.49 min; ESI-MS (+) m/z 952.4 (M+2H).

Analysis conditions B: residence time = 2.81 min; ESI-MS (+) m/z 952.5 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10179

The crude material of Example 10179 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.74 min; ESI-MS (+) m/z 976.5 (M+2H).

Analysis conditions B: residence time = 3.12 min; ESI-MS (+) m/z 976.1 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10180

The crude material of Example 10180 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.79 min; ESI-MS (+) m/z 975.7 (M+2H).

Analysis conditions B: residence time = 3.37 min; ESI-MS (+) m/z 975.8 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10181

The crude material of Example 10181 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.5 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.77 min; ESI-MS (+) m/z 960.70 (M+2H).

Analysis condition B: residence time = 3.37 min; ESI-MS (+) m/z 960.75 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 10182

The crude material of Example 10182 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.80 min; ESI-MS (+) m/z 936.25 (M+2H).

Analysis conditions B: residence time = 3.47 min; ESI-MS (+) m/z 936.20 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10183

The crude material of Example 10183 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.72 min; ESI-MS (+) m/z 960.75 (M+2H).

Analysis condition B: residence time = 3.29 min; ESI-MS (+) m/z 960.75 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 10184

The crude material of Example 10184 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.5 mg and the purity was estimated to be 94% according to LCMS analysis.

Analysis conditions A: residence time = 1.74 min; ESI-MS (+) m/z 944.20 (M+2H).

Analysis conditions B: residence time = 3.37 min; ESI-MS (+) m/z 944.20 (M+2H).

ESI-HRMS (+) m/z: Calculated: 149.

Preparation of Example 1018.5

The crude material of Example 10185 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.6 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.71 min; ESI-MS (+) m/z 960.65 (M+2H).

Analysis condition B: residence time = 3.23 min; ESI-MS (+) m/z 960.70 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 10186

The crude material of Example 10186 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 31.7 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.72 min; ESI-MS (+) m/z 960.70 (M+2H).

Analysis condition B: residence time = 3.30 min; ESI-MS (+) m/z 960.70 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10187

The crude material of Example 10187 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 50.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.82 min; ESI-MS (+) m/z 960.70 (M+2H).

Analysis condition B: residence time = 3.42 min; ESI-MS (+) m/z 960.75 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 10188

The crude material of Example 10188 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.67 min; ESI-MS (+) m/z 943.65 (M+2H).

Analysis condition B: residence time = 3.18 min; ESI-MS (+) m/z 943.65 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10189

The crude material of Example 10189 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.50 min; ESI-MS (+) m/z 952.0 (M+2H).

Analysis conditions B: residence time = 2.74 min; ESI-MS (+) m/z 952.0 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10190

The crude material of Example 10190 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.77 min; ESI-MS (-) m/z 935.2 (M-2H).

Analysis condition B: residence time = 2.97 min; ESI-MS (-) m/z 934.3 (M-2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10191

The crude material of Example 10191 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 38.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.79 min; ESI-MS (-) m/z 934.1 (M-2H).

Analysis conditions B: residence time = 2.99 min; ESI-MS (+) m/z 936.1 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10192

The crude material of Example 10192 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.58 min; ESI-MS (-) m/z 942.1 (M-2H).

Analysis conditions B: residence time = 2.66 min; ESI-MS (+) m/z 943.7 (M+2H).

ESI-HRMS (+) m/z: Calculated: s.

Preparation of Example 1019.3

The crude material of Example 10193 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 51.5 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis condition A: residence time = 1.62 min; ESI-MS (-) m/z 941.6 (M-2H).

Analysis conditions B: residence time = 2.68 min; ESI-MS (+) m/z 943.7 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10194

The crude material of Example 10194 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.82 min; ESI-MS (-) m/z 942.3 (M-2H).

Analysis conditions B: residence time = 2.97 min; ESI-MS (+) m/z 943.9 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 1019.5

The crude material of Example 10195 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.88 min; ESI-MS (-) m/z 967.4 (M-2H).

Analysis conditions B: residence time = 3.01 min; ESI-MS (+) m/z 968.8 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10196

The crude material of Example 10196 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 5.3 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.61 min; ESI-MS (+) m/z 895.7 (M+2H).

Analysis conditions B: residence time = 2.28 min; ESI-MS (+) m/z 895.2 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10197

The crude material of Example 10197 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.24 min; ESI-MS (+) m/z 902.15 (M+2H).

Analysis condition B: residence time = 1.83 min; ESI-MS (+) m/z 902.10 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10198

The crude material of Example 10198 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.24 min; ESI-MS (+) m/z 909.05 (M+2H).

Analysis conditions B: residence time = 2.58 min; ESI-MS (+) m/z 909.05 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10199

The crude material of Example 10199 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 35.4 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.61 min; ESI-MS (+) m/z 903.4 (M+2H).

Analysis conditions B: residence time = 2.22 min; ESI-MS (+) m/z 903.2 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10200

The crude material of Example 10200 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.7 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.47 min; ESI-MS (+) m/z 910.3 (M+2H).

Analysis conditions B: residence time = 2.15 min; ESI-MS (+) m/z 910.3 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10201

The crude material of Example 10201 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 45.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.54 min; ESI-MS (+) m/z 917.4 (M+2H).

Analysis conditions B: residence time = 2.22 min; ESI-MS (+) m/z 917.7 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10202

The crude material of Example 10202 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.45 min; ESI-MS (+) m/z 888.3 (M+2H).

Analysis conditions B: residence time = 2.18 min; ESI-MS (+) m/z 888.5 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 1020.3

The crude material of Example 10203 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.8 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z 895.6 (M+2H).

Analysis conditions B: residence time = 2.12 min; ESI-MS (+) m/z 895.3 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10204

The crude material of Example 10204 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.8 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.46 min; ESI-MS (+) m/z 902.7 (M+2H).

Analysis conditions B: residence time = 2.11 min; ESI-MS (+) m/z 902.3 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10205

The crude material of Example 10205 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.5 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.51 min; ESI-MS (+) m/z 896.1 (M+2H).

Analysis conditions B: residence time = 2.12 min; ESI-MS (+) m/z 896.4 (M+2H).

ESI-HRMS (+) m/z: Calcd.: 495.

Preparation of Example 10206

The crude material of Example 10206 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.42 min; ESI-MS (+) m/z 903.5 (M+2H).

Analysis conditions B: residence time = 2.07 min; ESI-MS (+) m/z 903.7 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10207

The crude material of Example 10207 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.2 mg and the purity was estimated to be 94% according to LCMS analysis.

Analysis conditions A: residence time = 1.46 min; ESI-MS (+) m/z 910.4 (M+2H).

Analysis condition B: residence time = 2.05 min; ESI-MS (+) m/z 910.4 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10208

The crude material of Example 10208 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 9.1 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.51 min; ESI-MS (+) m/z 881.9 (M+2H).

Analysis conditions B: residence time = 2.29 min; ESI-MS (+) m/z 882.1 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10209

The crude material of Example 10209 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.3 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.52 min; ESI-MS (+) m/z 882.3 (M+2H).

Analysis conditions B: residence time = 2.25 min; ESI-MS (+) m/z 882.4 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10210

The crude material of Example 10210 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.34 min; ESI-MS (+) m/z 874.50 (M+2H).

Analysis condition B: residence time = 1.95 min; ESI-MS (+) m/z 874.50 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10211

The crude material of Example 10211 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.57 min; ESI-MS (+) m/z 890.4 (M+2H).

Analysis conditions B: residence time = 2.29 min; ESI-MS (+) m/z 889.8 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10212

The crude material of Example 10212 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.56 min; ESI-MS (+) m/z 890.6 (M+2H).

Analysis condition B: residence time = 2.60 min; ESI-MS (+) m/z 890.35 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10213

The crude material of Example 10213 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.48 min; ESI-MS (+) m/z 882.7 (M+2H).

Analysis conditions B: residence time = 2.26 min; ESI-MS (+) m/z 882.8 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10214

The crude material of Example 10214 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.8 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.31 min; ESI-MS (+) m/z 888.80 (M+2H).

Analysis conditions B: residence time = 2.72 min; ESI-MS (+) m/z 888.85 (M+2H).

ESI-HRMS (+) m/z: calcd.

Preparation of Example 10215

The crude material of Example 10215 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.9 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.46 min; ESI-MS (+) m/z 889.4 (M+2H).

Analysis condition B: residence time = 2.24 min; ESI-MS (+) m/z 889.6 (M+2H).

ESI-HRMS (+) m/z: calc.: 189.

Preparation of Example 10216

The crude material of Example 10216 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.8 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.31 min; ESI-MS (+) m/z 881.45 (M+2H).

Analysis conditions B: residence time = 2.72 min; ESI-MS (+) m/z 881.70 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10217

The crude material of Example 10217 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.29 min; ESI-MS (+) m/z 896.60 (M+2H).

Analysis conditions B: residence time = 2.65 min; ESI-MS (+) m/z 896.55 (M+2H).

Preparation of Example 10218

The crude material of Example 10218 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.46 min; ESI-MS (-) m/z 895.30 (M-2H).

Analysis condition B: residence time = 2.20 min; ESI-MS (-) m/z 895.60 (M-2H).

ESI-HRMS (+) m/z: calc.: 896. s.

Preparation of Example 10219

The crude material of Example 10219 was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.29 min; ESI-MS (-) m/z 889.45 (M-2H).

Analysis condition B: residence time = 1.90 min; ESI-MS (-) m/z 890.35 (M-2H).

ESI-HRMS (+) m/z: Calcd.

Preparation of Example 10530

Example 10530 was prepared following "general synthesis sequence A" but using "cyclization method B" instead of "cyclization method A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 25-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation.

The yield of the product was 1.3 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 2.01 min; ESI-MS (+) m/z 915.4 (M+2H).

Analysis condition B: residence time = 2.94 min; ESI-MS (+) m/z 915.8 (M+2H.)

Preparation of Example 10531

Example 10531 was prepared following "general synthesis sequence A", but using "cyclization method B" instead of "cyclization method A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.56 min; ESI-MS (+) m/z 922.7 (M+2H).

Analysis conditions B: residence time = 2.94 min; ESI-MS (+) m/z 922.5 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10532

Example 10532 was prepared following "general synthesis sequence A" but using "cyclization method B" instead of "cyclization method A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.8 mg, and The purity estimate was 98% based on LCMS analysis.

Analysis conditions A: residence time = 1.77 min; ESI-MS (+) m/z 887.2 (M+2H).

Analysis condition B: residence time = 3.16 min; ESI-MS (+) m/z 887.2 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10533

Example 10533 was prepared following "General Synthetic Sequence A" with the following changes: "Symphony Method A: Single Coupling Procedure" was modified to extend the stirring time for the coupling of the Fmoc-Val-OH coupling step from 15 minutes to 3 hour. The Fmoc-Phe-OH coupling step uses "Symphony Method A: Single Coupling Procedure", but during the coupling step, the stirring time is extended from 15 min to 1 hour. "Overall removal of protecting group method A" was modified as follows: "Removal of protecting group solution" from [trifluoroacetic acid (22 mL), phenol (1.325 g), water (1.25 mL) and triisopropyl decane (0.5 mL) The solution was replaced with [trifluoroacetic acid (23.75 mL), 1,4-dithio-DL-threitol (625 mg), triisopropyl decane (0.625 mL)] liquid. Replace "Circularization Method A" with "Circularization Method B". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.70 min; ESI-MS (+) m/z 905.9 (M+2H).

Analysis conditions B: residence time = 3.27 min; ESI-MS (+) m/z 905.9 (M+2H).

Preparation of Example 10534

Example 10534 was prepared following "general synthesis sequence A" but using "cyclization method B" instead of "cyclization method A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-85% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.56 min; ESI-MS (+) m/z 951.5 (M+2H).

Analysis conditions B: residence time = 2.94 min; ESI-MS (+) m/z 951.4 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10535

Example 10535 was prepared according to the method used in Example 10533. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.76 min; ESI-MS (+) m/z 932.0 (M+2H).

Analysis conditions B: residence time = 3.34 min; ESI-MS (+) m/z 931.9 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10536

Example 10536 was prepared following "general synthesis sequence A" but using "cyclization method B" instead of "cyclization method A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 25-65% B for 30 minutes, followed by 100% B for 3 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 2.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.35 min; ESI-MS (+) m/z 927.8 (M+2H).

Analysis conditions B: residence time = 2.62 min; ESI-MS (+) m/z 927.6 (M+2H).

Preparation of Example 10539

Example 10539 was prepared following "general synthesis sequence A" but using "cyclization method B" instead of "cyclization method A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water + 0.1% trifluoroacetic acid; gradient: 25-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.2 mg, and the root According to LCMS analysis, the purity was estimated to be 96%.

Analysis conditions A: residence time = 1.19 min; ESI-MS (+) m/z 906.3 (M+2H).

Analysis conditions B: residence time = 2.32 min; ESI-MS (+) m/z 906.6 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10540

Example 10540 was prepared following "general synthesis sequence A" but using "cyclization method B" instead of "cyclization method A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, It was then held at 100% B for 3 minutes; flow rate: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.71 min; ESI-MS (+) m/z 934.6 (M+2H).

Analysis conditions B: residence time = 2.52 min; ESI-MS (+) m/z 934.3 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10542

Example 10542 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 150 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 3.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.92 min; ESI-MS (+) m/z 946.4 (M+2H).

Analysis conditions B: residence time = 2.91 min; ESI-MS (+) m/z 946.4 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10543

Example 10543 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.0 mg and the purity was estimated to be 93% according to LCMS analysis.

Analysis conditions A: residence time = 1.67 min; ESI-MS (+) m/z 919.9 (M+2H).

Analysis condition B: residence time = 3.26 min; ESI-MS (+) m/z 919.9 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10544

Example 10544 was prepared following "general synthesis sequence A" but using "cyclization method B" instead of "cyclization method A". The crude material was purified by preparative LC/MS using the following conditions. Tube: column: XBridge C18, 19 × 200mm, 5μm particles; mobile phase A: 5:95 methanol: water + 10mM ammonium acetate; mobile phase B: 95:5 methanol: water + 10mM ammonium acetate; gradient: 35-75 % B lasted 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.4 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.36 min; ESI-MS (+) m/z 936.4 (M+2H).

Analysis conditions B: residence time = 2.14 min; ESI-MS (+) m/z 936.4 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10551 and Example 10640

Example 10551 and Example 10640 were prepared according to the method outlined in Example 10541, but in which the dipeptide fragment Fmoc-L-Val-L- ^m Phe-OH was used in place of the dipeptide fragment Fmoc-L-cyclopentyl Gly-L- ^m Phe -OH. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The diastereomers were separated and dried via EtOAc (EtOAc: EtOAc: EtOAc: EtOAc: EtOAc: According to LCMS analysis, the purity was estimated to be 98%.

Example 10551:

Analysis conditions A: residence time = 1.67 min; ESI-MS (+) m/z 966.9 (M+2H).

Analysis condition B: residence time = 3.28 min ; ESI-MS (+) m/z 966.9 (M+2H).

ESI-HRMS (+) m/z : calc.

Example 10640:

Analysis conditions A: residence time = 1.78 min; ESI-MS (+) m/z 967.0 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10552

Example 10552 was prepared following "general synthesis sequence A" but using "cyclization method B" instead of "cyclization method A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.7 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.70 min; ESI-MS (+) m/z 956.9 (M+2H).

Analysis conditions B: residence time = 2.47 min; ESI-MS (+) m/z 956.9 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10553

Example 10553 was prepared following "general synthesis sequence A" but using "cyclization method B" instead of "cyclization method A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.55 min ; ESI-MS (+) m/z 928.2 (M+2H).

Analysis conditions B: residence time = 2.21 min; ESI-MS (+) m/z 928.1 (M+2H).

ESI-HRMS (+) m/z : calc.: 927.

Preparation of Example 10554

Example 10554 was prepared following "general synthesis sequence A" but using "cyclization method B" instead of "cyclization method A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 25-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.8 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.87 min; ESI-MS (+) m/z 948.8 (M+2H).

Analysis conditions B: residence time = 2.72 min; ESI-MS (+) m/z 948.3 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10555

Example 10555 was prepared following "general synthesis sequence A" but using "cyclization method B" instead of "cyclization method A". The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.89 min; ESI-MS (+) m/z 955.3 (M+2H).

Analysis conditions B: residence time = 2.75 min; ESI-MS (+) m/z 955.6 (M+2H).

ESI-HRMS (+) m/z : Calcd.

Preparation of Example 10556

Example 10556 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 60-100% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 18.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.94 min; ESI-MS (+) m/z 958.8 (M+2H).

Analysis conditions B: residence time = 2.78 min; ESI-MS (+) m/z 958.8 (M+2H).

Preparation of Example 10557

Example 10557 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 60-100% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.94 min; ESI-MS (+) m/z 946.0 (M+2H).

Analysis conditions B: residence time = 2.77 min; ESI-MS (+) m/z 946.0 (M+2H).

Preparation of Example 10558

Example 10558 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 7.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.93 min; ESI-MS (+) m/z 958.9 (M+2H).

Analysis conditions B: residence time = 2.75 min; ESI-MS (+) m/z 958.9 (M+2H).

Preparation of Example 10559

Example 10559 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.92 min; ESI-MS (+) m/z 965.9 (M+2H).

Analysis conditions B: residence time = 2.77 min; ESI-MS (+) m/z 966.1 (M+2H).

Preparation of Example 10560

Example 10560 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 60-100% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 35.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 2.08 min; ESI-MS (+) m/z 966.0 (M+2H).

Analysis conditions B: residence time = 2.87 min; ESI-MS (+) m/z 966.1 (M+2H).

Preparation of Example 10561

Example 10561 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.2 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 2.14 min; ESI-MS (+) m/z 951.4 (M+2H).

Analysis conditions B: residence time = 2.85 min; ESI-MS (+) m/z 950.6 (M+2H).

Preparation of Example 10562

Example 10562 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.80 min; ESI-MS (+) m/z 951.0 (M+2H).

Analysis condition B: residence time = 3.32 min; ESI-MS (+) m/z 950.3 (M+2H).

Preparation of Example 1056.3

Example 10563 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 32.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.77 min; ESI-MS (+) m/z 958.1 (M+2H).

Analysis conditions B: residence time = 3.33 min; ESI-MS (+) m/z 958.4 (M+2H).

Preparation of Example 10564

Example 10564 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.88 min; ESI-MS (+) m/z 957.9 (M+2H).

Analysis conditions B: residence time = 2.52 min; ESI-MS (+) m/z 957.9 (M+2H).

Preparation of Example 10565

Example 10565 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.7 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.84 min; ESI-MS (+) m/z 937.6 (M+2H).

Analysis conditions B: residence time = 3.38 min; ESI-MS (+) m/z 937.6 (M+2H).

Preparation of Example 10566

Example 10566 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.1 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.70 min; ESI-MS (+) m/z 952.4 (M+2H).

Analysis conditions B: residence time = 3.36 min; ESI-MS (+) m/z 952.5 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10567

Example 10567 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 15.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.70 min; ESI-MS (+) m/z 945.4 (M+2H).

Analysis conditions B: residence time = 3.35 min; ESI-MS (+) m/z 945.4 (M+2H).

ESI-HRMS (+) m/z : Calculated: </RTI></RTI></RTI><RTIgt;

Preparation of Example 10568

Example 10568 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.8 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.81 min; ESI-MS (+) m/z 944.5 (M+2H).

Analysis conditions B: residence time = 3.45 min; ESI-MS (+) m/z 944.5 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10569

Example 10569 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.3 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.79 min; ESI-MS (+) m/z 937.2 (M+2H).

Analysis conditions B: residence time = 3.39 min; ESI-MS (+) m/z 937.6 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10570

Example 10570 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.77 min; ESI-MS (+) m/z 944.0 (M+2H).

Analysis condition B: residence time = 3.39 min; ESI-MS (+) m/z 943.9 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10571

Example 10571 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.7 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.83 min; ESI-MS (+) m/z 937.2 (M+2H).

Analysis conditions B: residence time = 3.44 min; ESI-MS (+) m/z 937.4 (M+2H).

ESI-HRMS (+) m/z : </RTI> : < / RTI></RTI><RTIgt;

Preparation of Example 10572

Example 10572 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.9 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.76 min; ESI-MS (+) m/z 930.0 (M+2H).

Analysis condition B: residence time = 3.31 min; ESI-MS (+) m/z 930.4 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10573

Example 10573 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.8 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.72 min; ESI-MS (+) m/z 930.8 (M+2H).

Analysis conditions B: residence time = 3.25 min; ESI-MS (+) m/z 930.9 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10574

Example 10574 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.1 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.71 min; ESI-MS (+) m/z 944.4 (M+2H).

Analysis condition B: residence time = 3.24 min; ESI-MS (+) m/z 944.3 (M+2H).

ESI-HRMS (+) m/z : Calculated: < / RTI></RTI></RTI></RTI><RTIgt;

Preparation of Example 10575

Example 10575 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.0 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.67 min; ESI-MS (+) m/z 951.4 (M+2H).

Analysis conditions B: residence time = 3.22 min; ESI-MS (+) m/z 951.4 (M+2H).

ESI-HRMS (+) m/z : Calculated: 950.9050 (M+2H);

Preparation of Example 10576

Example 10576 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 31.8 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.77 min; ESI-MS (+) m/z 957.9 (M+2H).

Analysis conditions B: residence time = 3.30 min; ESI-MS (+) m/z 957.9 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10577

Example 10577 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.0 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.94 min; ESI-MS (+) m/z 943.4 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10078

Example 10078 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.3 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.95 min; ESI-MS (+) m/z 936.3 (M+2H).

ESI-HRMS (+) m / z: Calcd: 935.9361 (M + 2H); Found: 935.9335 (M + 2H).

Preparation of Example 10579

Example 10579 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.9 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.94 min; ESI-MS (+) m/z 950.1 (M+2H).

Analysis conditions B: residence time = 3.46 min; ESI-MS (+) m/z 950.1 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10580

Example 10580 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.2 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.81 min; ESI-MS (+) m/z 951.0 (M+2H).

Analysis conditions B: residence time = 3.34 min; ESI-MS (+) m/z 950.1 (M+2H).

ESI-HRMS (+) m/z : Calcd.

Preparation of Example 10581

Example 10581 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 39.7 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.94 min; ESI-MS (+) m/z 936.3 (M+2H).

Analysis conditions B: residence time = 3.46 min; ESI-MS (+) m/z 936.4 (M+2H).

ESI-HRMS (+) m/z : </RTI> : < / RTI></RTI></RTI><RTIgt;

Preparation of Example 1058.2

Example 10582 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 27.1 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.95 min; ESI-MS (+) m/z 929.3 (M+2H).

ESI-HRMS (+) m/z : Calculated: < / RTI></RTI></RTI></RTI><RTIgt;

Preparation of Example 10583

Example 10583 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.8 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.95 min; ESI-MS (+) m/z 929.9 (M+2H).

Analysis conditions B: residence time = 3.46 min; ESI-MS (+) m/z 929.9 (M+2H).

Preparation of Example 10584

Example 10584 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.3 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.91 min; ESI-MS (+) m/z 943.4 (M+2H).

Analysis conditions B: residence time = 3.43 min; ESI-MS (+) m/z 943.4 (M+2H).

Preparation of Example 10585

Example 10585 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.0 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis condition A: retention time = 1.48min; ESI-MS (+ ) m / z 972.3 (M + 2H).

Analysis conditions B: residence time = 3.12 min; ESI-MS (+) m/z 972.5 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10586

Example 10586 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 28.7 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.41 min; ESI-MS (+) m/z 978.7 (M+2H).

Analysis conditions B: residence time = 2.77 min; ESI-MS (+) m/z 978.7 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10587

Example 10587 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.4 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.98 min; ESI-MS (+) m/z 936.4 (M+2H).

Analysis conditions B: residence time = 3.46 min; ESI-MS (+) m/z 936.4 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10588

Example 10588 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5; 95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.98 min; ESI-MS (-) m/z 949.9 (M-2H).

Analysis conditions B: residence time = 2.93 min; ESI-MS (+) m/z 951.9 (M+2H).

Preparation of Example 10589

Example 10589 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.1 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.70 min; ESI-MS (+) m/z 944.9 (M+2H).

Analysis conditions B: residence time = 2.88 min; ESI-MS (+) m/z 944.9 (M+2H).

Preparation of Example 10590

Example 10590 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-60% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.4 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.71 min; ESI-MS (+) m/z 959.2 (M+2H).

Analysis conditions B: residence time = 2.90 min; ESI-MS (+) m/z 958.7 (M+2H).

Preparation of Example 10591

Example 10591 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 8.2 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis condition A: residence time = 1.71 min; ESI-MS (-) m/z 957.5 (M-2H).

Analysis condition B: residence time = 2.90 min; ESI-MS (-) m/z 957.5 (M-2H).

Preparation of Example 10592

Example 10592 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 13.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.70 min; ESI-MS (+) m/z 966.3 (M+2H).

Analysis conditions B: residence time = 2.91 min; ESI-MS (+) m/z 966.2 (M+2H).

Preparation of Example 10593

Example 10593 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.70 min; ESI-MS (-) m/z 949.5 (M-2H).

Analysis conditions B: residence time = 2.87 min; ESI-MS (+) m/z 952.0 (M+2H).

Preparation of Example 10594

Example 10594 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 35-75% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-65% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.2 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.73 min; ESI-MS (+) m/z 958.9 (M+2H).

Analysis conditions B: residence time = 3.3 min; ESI-MS (+) m/z 958.9 (M+2H).

Preparation of Example 10595

Example 10595 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 6.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.59 min; ESI-MS (+) m/z 966.2 (M+2H).

Analysis conditions B: residence time = 2.77 min; ESI-MS (+) m/z 966.0 (M+2H).

Preparation of Example 10596

Example 10596 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.80 min; ESI-MS (-) m/z 951.0 (M-2H).

Analysis conditions B: residence time = 2.97 min; ESI-MS (+) m/z 952.4 (M+2H).

Preparation of Example 10597

Example 10597 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 19.2 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.73 min; ESI-MS (-) m/z 950.5 (M-2H).

Analysis conditions B: residence time = 2.93 min; ESI-MS (+) m/z 952.2 (M+2H).

Preparation of Example 10598

Example 10598 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 50-100% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 24.5 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.75 min; ESI-MS (+) m/z 959.5 (M+2H).

Analysis conditions B: residence time = 2.97 min; ESI-MS (+) m/z 959.7 (M+2H).

Preparation of Example 10599

Example 10599 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 34.2 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.77 min; ESI-MS (-) m/z 948.0 (M-2H).

Analysis conditions B: residence time = 2.82 min; ESI-MS (+) m/z 949.6 (M+2H).

Preparation of Example 10600

Example 10600 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.75 min; ESI-MS (-) m/z 942.6 (M-2H).

Analysis conditions B: residence time = 2.98 min; ESI-MS (+) m/z 943.5 (M+2H).

Preparation of Example 10601

Example 10601 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.67 min; ESI-MS (+) m/z 950.8 (M+2H).

Analysis conditions B: residence time = 2.98 min; ESI-MS (+) m/z 950.6 (M+2H).

Preparation of Example 10602

Example 10602 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.18 min; ESI-MS (-) m/z 955.6 (M-2H).

Analysis conditions B: residence time = 2.84 min; ESI-MS (+) m/z 956.9 (M+2H).

Preparation of Example 10603

Example 10603 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 31.9 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.75 min; ESI-MS (+) m/z 950.5 (M+2H).

Analysis condition B: retention time = 2.96min; ESI-MS (+ ) m / z 949.9 (M + 2H).

Preparation of Example 10604

Example 10604 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.3 mg and the purity was estimated to be 94% according to LCMS analysis.

Analysis conditions A: residence time = 1.74 min; ESI-MS (-) m/z 934.5 (M-2H).

Analysis conditions B: residence time = 2.95 min; ESI-MS (+) m/z 936.7 (M+2H).

Preparation of Example 10605

Example 10605 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 55-95% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 30-70% B for 15 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.80 min; ESI-MS (-) m/z 943.0 (M-2H).

Analysis conditions B: residence time = 3.06 min; ESI-MS (+) m/z 945.5 (M+2H).

Preparation of Example 10606

Example 10606 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 30-70% B for 20 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.6 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.73 min; ESI-MS (+) m/z 938.4 (M+2H).

Analysis conditions B: residence time = 2.94 min; ESI-MS (+) m/z 938.5 (M+2H).

Preparation of Example 10607

Example 10607 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. Production The yield was 3.4 mg and the purity was estimated to be 95% according to LCMS analysis.

Analysis conditions A: residence time = 1.62 min; ESI-MS (+) m/z 959.5 (M+2H).

Analysis conditions B: residence time = 2.97 min; ESI-MS (+) m/z 959.3 (M+2H).

ESI-HRMS (+) m/z : calc.: 958.

Preparation of Example 10608

Example 10608 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 39.9 mg, and according to LCMS The purity is estimated to be 100%.

Analysis conditions A: residence time = 1.57 min; ESI-MS (-) m/z 920.5 (M-2H).

Analysis condition B: residence time = 3.10 min; ESI-MS (+) m/z 922.5 (M+2H).

ESI-HRMS (+) m / z: Calcd: 922.4048 (M + 2H); Found: 922.4025 (M + 2H).

Preparation of Example 10609

Example 10609 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 20 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.7 mg, and according to LCMS The purity is estimated to be 99%.

Analysis conditions A: residence time = 1.60 min; ESI-MS (+) m/z 908.4 (M+2H).

Analysis conditions B: residence time = 3.06 min; ESI-MS (+) m/z 908.4 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10610

Example 10610 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 5-45% B for 20 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 26.8 mg, and according to LCMS The purity was estimated to be 96%.

Analysis condition A: residence time = 1.63 min; ESI-MS (-) m/z 937.4 (M-2H).

Analysis condition B: residence time = 2.25 min; ESI-MS (-) m/z 936.5 (M-2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10611

Example 10611 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 17.7 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.40 min; ESI-MS (-) m/z 973.4 (M-2H).

Analysis conditions B: residence time = 2.64 min; ESI-MS (+) m/z 975.4 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10612

Example 10612 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 11.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis condition A: residence time = 1.58 min; ESI-MS (-) m/z 958.1 (M- 2H).

Analysis conditions B: residence time = 2.92 min; ESI-MS (+) m/z 960.5 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10613

Example 10613 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 12.2 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.76 min; ESI-MS (+) m/z 945.4 (M+2H).

Analysis conditions B: residence time = 3.20 min; ESI-MS (+) m/z 945.7 (M+2H).

ESI-HRMS (+) m/z : Calculated: 149.

Preparation of Example 10614

Example 10614 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was further purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM acetic acid Ammonium; Gradient: 40-80% B for 20 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 16.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition A: residence time = 1.59 min; ESI-MS (-) m/z 965.6 (M-2H).

Analysis conditions B: residence time = 2.90 min; ESI-MS (+) m/z 967.4 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10615

Example 10615 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B lasts 30 minutes, then 100% B for 5 minutes; flow rate: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 21.1 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis condition A: residence time = 1.72 min; ESI-MS (-) m/z 948.6 (M-2H).

Analysis condition B: residence time = 2.88 min; ESI-MS (-) m/z 948.0 (M-2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10616

Example 10616 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. Combine the fractions containing the desired product It was dried by centrifugal evaporation. The yield of the product was 29.3 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis condition A: residence time = 1.74 min; ESI-MS (-) m/z 942.0 (M-2H).

Analysis condition B: residence time = 2.99 min; ESI-MS (-) m/z 942.0 (M-2H).

ESI-HRMS (+) m/z : Calculated: < / RTI></RTI></RTI><RTIgt;

Preparation of Example 10617

Example 10617 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. Combine the fractions containing the desired product It was dried by centrifugal evaporation. The yield of the product was 18.6 mg and its purity was estimated to be 100% according to LCMS analysis.

Analysis conditions B: residence time = 2.78 min; ESI-MS (-) m/z 934.4 (M-2H).

Preparation of Example 10618

Example 10618 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 37.3 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.73 min; ESI-MS (+) m/z 951.2 (M+2H).

Analysis conditions B: residence time = 2.98 min; ESI-MS (+) m/z 950.4 (M+2H).

Preparation of Example 10619

Example 10619 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.65 min; ESI-MS (-) m/z 955.5 (M-2H).

Analysis conditions B: residence time = 2.84 min; ESI-MS (+) m/z 957.6 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10620

Example 10620 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.68 min; ESI-MS (-) m/z 656.6 (M-2H).

Analysis conditions B: residence time = 2.73 min; ESI-MS (-) m/z 955.9 (M-2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10621

Example 10621 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 27.5 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.62 min; ESI-MS (-) m/z 957.6 (M-2H).

Analysis conditions B: residence time = 2.82 min; ESI-MS (+) m/z 958.7 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10622

Example 10622 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 35.6 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.92 min; ESI-MS (+) m/z 936.5 (M+2H).

Analysis condition B: residence time = 3.11 min; ESI-MS (-) m/z 933.5 (M-2H).

Preparation of Example 10623

Example 10623 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 39.9 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.90 min; ESI-MS (+) m/z 930.0 (M+2H).

Analysis condition B: residence time = 3.11 min; ESI-MS (-) m/z 927.0 (M-2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10624

Example 10624 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 27.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions B: residence time = 2.98 min; ESI-MS (+) m/z 943.4 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10625

Example 10625 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 30.6 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.89 min; ESI-MS (+) m/z 936.5 (M+2H).

Analysis conditions B: residence time = 3.11 min; ESI-MS (+) m/z 935.9 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10626

Example 10626 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 20-60% B for 20 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 4.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.82 min; ESI-MS (-) m/z 942.5 (M-2H).

Analysis conditions B: residence time = 3.08 min; ESI-MS (+) m/z 943.6 (M+2H).

Preparation of Example 10627

Example 10627 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.9 mg and the purity was estimated to be 97% according to LCMS analysis.

Analysis conditions A: residence time = 1.79 min; ESI-MS (+) m/z 944.9 (M+2H).

Analysis conditions B: residence time = 3.04 min; ESI-MS (+) m/z 944.1 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10628

Example 10628 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 32.3 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.70 min; ESI-MS (-) m/z 950.8 (M-2H).

Analysis conditions B: residence time = 2.91 min; ESI-MS (+) m/z 951.6 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10629

Example 10629 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 14.0 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions B: residence time = 2.85 min; ESI-MS (+) m/z 950.0 (M+2H).

Analysis conditions C: residence time = 1.86 min; ESI-MS (+) m/z 950.7 (M+2H).

ESI-HRMS (+) m/z : Calcd.

Preparation of Example 10630

Example 10630 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 36.4 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.64 min; ESI-MS (-) m/z 949.0 (M-2H).

Analysis conditions B: residence time = 2.79 min; ESI-MS (+) m/z 950.6 (M+2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10631

Example 10631 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 23.2 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.57 min; ESI-MS (-) m/z 955.9 (M-2H).

Analysis condition B: residence time = 2.79 min; ESI-MS (-) m/z 956.1 (M-2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10632

Example 10632 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 15-55% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 20.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.64 min; ESI-MS (-) m/z 949.2 (M-2H).

Analysis conditions B: residence time = 2.89 min; ESI-MS (-) m/z 948.4 (M-2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10633

Example 10633 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 40-80% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 27.8 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.56 min; ESI-MS (-) m/z 957.4 (M-2H).

Analysis condition B: residence time = 2.67 min; ESI-MS (-) m/z 956.0 (M-2H).

ESI-HRMS (+) m/z : calcd.

Preparation of Example 10634

Example 10634 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 32.1 mg and the purity was estimated to be 100% according to LCMS analysis.

Analysis conditions A: residence time = 1.61 min; ESI-MS (-) m/z 956.7 (M-2H).

Analysis conditions B: residence time = 2.74 min; ESI-MS (+) m/z 958.2 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10635

Example 10635 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile : water + 10 mM ammonium acetate; gradient: 10-50% B for 30 minutes, then held at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 25.3 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.53 min; ESI-MS (-) m/z 956.6 (M-2H).

Analysis conditions B: residence time = 2.69 min; ESI-MS (+) m/z 959.5 (M+2H).

ESI-HRMS (+) m/z : calc.

Preparation of Example 10636

Example 10636 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 22.0 mg and the purity was estimated to be 96% according to LCMS analysis.

Analysis conditions A: residence time = 1.51 min; ESI-MS (-) m/z 964.5 (M-2H).

Analysis conditions B: residence time = 2.69 min; ESI-MS (+) m/z 965.6 (M+2H).

ESI-HRMS (+) m/z : Calculated: </RTI></RTI>

Preparation of Example 10637

Example 10637 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 45-85% B for 30 minutes, then at 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 35.4 mg and the purity was estimated to be 98% according to LCMS analysis.

Analysis conditions A: residence time = 1.51 min; ESI-MS (+) m/z 966.5 (M+2H).

Analysis conditions B: residence time = 2.57 min; ESI-MS (+) m/z 966.6 (M+2H).

ESI-HRMS (+) m/z : calc.: s.

Preparation of Example 10638

Example 10638 was prepared according to the method outlined in Example 10537. The crude material was purified via preparative LC/MS using the following conditions: column: XBridge C18, 19 x 200 mm, 5 μm particles; mobile phase A: 5:95 methanol: water + 10 mM ammonium acetate; mobile phase B: 95:5 methanol : water + 10 mM ammonium acetate; gradient: 50-90% B for 30 minutes, followed by 100% B for 5 minutes; flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 29.6 mg and the purity was estimated to be 99% according to LCMS analysis.

Analysis conditions A: residence time = 1.63 min; ESI-MS (-) m/z 956.9 (M-2H).

Analysis condition B: residence time = 2.81 min; ESI-MS (-) m/z 956.5 (M-2H).

ESI-HRMS (+) m/z : calcd.

Analytical LCMS conditions XX:

Column: Waters Xbridge C18, 2.1×50 mm; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; temperature: 35 ° C; gradient: 0- 100% B lasted 8 minutes, then held at 100% B for 1 minute; flow rate: 0.8 mL/min; detection: 220 nm UV.

Preparation of Example 11001

Example 11001 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method A: Resin Swelling Procedure ", "Symphony Method A: Standard Coupling Procedure ", "Symphony Method" A: secondary amine coupling procedure B ", " custom amino acid coupling procedure ", " chloroacetic acid coupling procedure B ", " total removal of protecting group method C " and " cyclization method D ". (S)-2-(((9H-茀-9-yl)methoxy)carbonyl))amino)-3,3-dimethylbutyric acid is used in the " custom amino acid coupling procedure ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 30 x 100 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 15-55% B for 25 minutes, followed by 100% B for 5 minutes; flow: 30 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 10.2 mg and the purity was estimated to be 98.2% according to LCMS analysis.

Analytical LCMS conditions XX: retention time = 4.66 min; ESI-MS (+) m/z 949.52 (M+2H).

Preparation of Example 11002

Example 11002 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method A: Resin Swelling Procedure ", "Symphony Method A: Standard Coupling Procedure ", "Symphony Method" A: secondary amine coupling procedure B ", " custom amino acid coupling procedure ", " chloroacetic acid coupling procedure B ", " total removal of protecting group method C " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 30 x 100 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 15-55% B for 25 minutes, then at 100% B for 5 minutes; flow: 30 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 38 mg and the purity was estimated to be 93.6% according to LCMS analysis.

Analytical LCMS conditions XX: retention time = 3.64 min; ESI-MS (+) m/z 941.70 (M+2H).

ESI-HRMS (+) m/z: Calculated: 149.

Preparation of Example 11003

Example 11003 was prepared following the general synthetic sequence described for the preparation of Example 0001, which consisted of the following general procedure: "Symphony Method A: Resin Swelling Procedure ", "Symphony Method A: Standard Coupling Procedure ", "Symphony Method" A: secondary amine coupling procedure B ", " custom amino acid coupling procedure ", " chloroacetic acid coupling procedure B ", " total removal of protecting group method C " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 30 x 100 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 15-55% B for 25 minutes, then at 100% B for 5 minutes; flow: 30 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 50 mg and the purity was estimated to be 95.6% according to LCMS analysis.

Analytical LCMS conditions XX: retention time = 3.55 min; ESI-MS (+) m/z 976.30 (M+2H).

ESI-HRMS (+) m/z: calc.

Preparation of Example 11004

Was prepared following the general synthetic sequences for Example 11 004 Example of the preparation of 0001, the general synthetic sequence consisting of the following general procedure: "Symphony Method A: resin swollen program", "Symphony Method A: Standard coupling procedures", "method Symphony A: secondary amine coupling procedure B ", " custom amino acid coupling procedure ", " chloroacetic acid coupling procedure B ", " total removal of protecting group method C " and " cyclization method D ".

The crude material was purified via preparative LC/MS using the following conditions: column: Waters XBridge C18, 30 x 100 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 Acetonitrile: water + 10 mM ammonium acetate; gradient: 15-55% B for 25 minutes, followed by 100% B for 5 minutes; flow: 30 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of the product was 70 mg and the purity was estimated to be 96.5% based on LCMS analysis.

Analytical LCMS conditions XX: residence time = 3.78 min; ESI-MS (+) m/z 942.30 (M+2H).

ESI-HRMS (+) m/z: Calcd.

Method for testing the ability of macrocyclic peptides to compete with PD-1 for binding to PD-L1 using homogenous time-lapse fluorescence (HTRF) binding assay

The present invention was studied using PD-1/PD-L1 homogeneous time difference fluorescence (HTRF) binding assay The ability of macrocyclic peptides to bind to PD-L1.

method

Homogeneous Time Difference Fluorescence (HTRF) analysis of soluble PD-1 binding to soluble PD-L1. Soluble PD-1 and soluble PD-L1 are proteins with a carboxy-terminal truncation that are truncated to remove the transmembrane region and fused to a heterologous sequence, particularly to the Fc portion of the human immunoglobulin G sequence (Ig). Or hexameric histidine epitope tag (His). All binding studies were performed using HTRF assay buffer consisting of dPBS supplemented with 0.1% (w/v) bovine serum albumin and 0.05% (v/v) Tween-20. In the PD-1-Ig/PD-L1-His binding assay, the inhibitor was pre-incubated with PD-L1-His (10 nM final concentration) in 4 μl of assay buffer for 15 minutes, followed by the addition of PD-1-Ig (20 nM). 1 μl of assay buffer at final concentration) and further incubated for 15 minutes. A PD-L1 fusion protein derived from human, cynomolgus monkey or mouse is used. HTRF detection was achieved using an anti-Ig monoclonal antibody (1 nM final concentration) labeled with a cryptate compound and an allophycocyanin (APC)-labeled anti-His monoclonal antibody (20 nM final concentration). The antibody was diluted in HTRF detection buffer and 5 [mu]l was dispensed onto the binding reaction. The reaction was allowed to equilibrate for 30 minutes and the signal was obtained using an EnVision fluorometer (665 nm / 620 nm ratio). Establish PD-1-Ig/PD-L2-His (20nM and 5nM, respectively), CD80-His/PD-L1-Ig (100nM and 10nM, respectively) and CD80-His/CTLA4-Ig (10nM and 5nM, respectively) ) Other combinations of analysis between. A competition study between biotinylated compound No. 71 and human PD-L1-His was performed as follows. The macrocyclic peptide inhibitor was preincubated with PD-L1-His (10 nM final) in 4 μl of assay buffer for 60 minutes, followed by the addition of 1 μl of assay buffer containing biotinylated compound No. 71 (0.5 nM final concentration). Binding was allowed for 30 minutes, followed by the addition of 5 μl of HTRF buffer containing streptavidin-labeled streptavidin (2.5 pM final concentration) and APC-labeled anti-His (20 nM final concentration). The reaction was allowed to equilibrate for 30 minutes and the signal was obtained using an EnVision fluorometer (665 nm / 620 nm ratio).

Recombinant protein. Cleavage of human PD-1 (amino acid 25-167) [hPD-1(25-167)-3S-IG] with a C-terminal human Ig epitope tag and having a C-terminal His antigen The base-labeled human PD-L1 (amino acid 18-239) [hPD-L1(19-239)-tobacco vein plaque protease cleavage site (TVMV)-His] is expressed in HEK293T cells and sequentially by rProteinA affinity Purification by chromatography and size exclusion chromatography. Human PD-L2-His (Sino Biologicals), CD80-His (Sino Biologicals), and CTLA4-Ig (RnD Systems) were all obtained from commercial sources.

Recombinant human PD-1-Ig sequence

(SEQ ID NO : 1)

Recombinant human PD-L1-TVMV-His (PD-L1-His) sequence

(SEQ ID NO : 2)

The results are shown in Table 1. As shown, the macrocyclic peptide of the present invention showed strong inhibition of the activity of PD-1-Ig binding to PD-L1-TVMV-His (PD-L1-His). A = 0.001 - 0.0099 μM; B = 0.01 - 0.0999 μM; C = 0.10 - 0.99 μM; D = 1-10 μM.

The subject matter described and claimed herein can be modified and altered in light of the above teachings. Therefore, it is to be understood that within the scope of the appended claims, the subject matter described in the claims can be practiced otherwise.

The scope of the present invention is not limited by the embodiments disclosed herein, and such embodiments are intended to be a single description of the individual aspects of the invention, and any aspects that are functionally equivalent are within the scope of the invention. In addition to the teachings herein, various modifications of the models and methods of the present invention are readily apparent to those skilled in the art in light of the above description and teachings, and such modifications are also intended to be within the scope of the present invention. Such refinements or other embodiments may be practiced without departing from the true scope and spirit of the invention.

All references cited in this document (including patents, patent applications, journal articles, abstracts, laboratory manuals, books, GENBANK® registration numbers, SWISS-PROT® registration numbers or other disclosures) are all cited in full. The manner is incorporated herein. In addition, the papers accompanying the presented sequence listing, in addition to their respective computer readable forms, are also incorporated herein by reference in their entirety.

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<120> Macrocyclic inhibitors of PD-1/PD-L1 and CD80(B7-1)/PD-L1 protein/protein interactions

<130> 12386-WO-PCT

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<170> PatentIn version 3.5

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<211> 384

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Claims (13)

A compound selected from the group consisting of: Example 3210, Example 3210, Example 3211, Example 3212, Example 3213, Example 3216, Example 3217, Example 3218, Example 3219, Example 3220, Example 3221, Example 3222, Example 3223, Example 3224, Example 3225, instance 3226, instance 3227, instance 3228, instance 3229, instance 3230, instance 3231, instance 3232, instance 3233, instance 3234, instance 3235, instance 3236, instance 3237, instance 3238, instance 3239, instance 3240, instance 3241 Example 3242, Example 3243, Example 3244, Example 3245, Example 3246, Example 3614, Example 3616, Example 3617, Example 3618, Example 3628, Example 3637, Example 3638, Example 3639, Example 3640, Example 3641, Example 3642, Example 3643 Example 3644, Example 3645, Example 3646, Example 3647, Example 3648, Example 9166, Example 9167, Example 9188, Example 9189, Example 9190, Example 9191, Example 9112, Example 9193, Example 9194, Example 9195, Example 9196, Example 9213, instance 9214, instance 9215, instance 9216, instance 9217, instance 9218, instance 9219, instance 9220, instance 9221, instance 9222, instance 92 23. Example 9224, Example 9225, Example 9226, Example 9227, Example 9228, Example 9229, Example 9230, Example 9231, Example 9232, Example 9233, Example 9234, Example 9235, Example 9236, Example 9237, Example 9238, Example 9239, Example 9240, instance 9241, instance 9242, instance 9243, instance 9244, instance 9245, instance 9246, instance 9247, instance 9248, instance 9249, instance 9250, instance 9251, instance 9252, instance 9253, instance 9254, instance 9255, instance 9256 , instance 9257, instance 9258, instance 9259, instance 9260, instance 9261, instance 9262, instance 9263, instance 9264, instance 9265, instance 9266, instance 9267, instance 9268, instance 9269, instance 9270, instance 9271, instance 9272, instance 9273, instance 9274, instance 9275, instance 9276, instance 9277, instance 9278, instance 9279, instance 9280, Example 9281, instance 9282, instance 9283, instance 9284, instance 9285, instance 9286, instance 9287, instance 9288, instance 9289, instance 9290, instance 9291, instance 9292, instance 9293, instance 9294, instance 9295, instance 9296, instance 9297 , instance 9298, instance 9299, instance 9300, instance 9301, instance 9302, instance 9303, instance 9304, instance 9305, instance 9306, instance 9307, instance 9308, instance 9309, instance 9310, instance 9311, instance 9132, instance 9313, instance 9314, instance 9315, instance 9316, instance 9317, instance 9318, instance 9319, instance 9320, instance 9321, instance 9322, instance 9323, instance 9324, instance 9325, instance 9326, instance 9327, instance 9328, instance 9329, instance 9330, Example 9131, instance 9332, instance 9333, instance 9334, instance 9335, instance 9336, instance 9337, instance 9338, instance 9345, instance 9346, real 9347, Example 9348, Example 9349, Example 9350, Example 9351, Example 9352, Example 9353, Example 9354, Example 9355, Example 9356, Example 9357, Example 9358, Example 9359, Example 9360, Example 9361, Example 9362, Example 9363, Example 9364, instance 9365, instance 9366, instance 9367, instance 9368, instance 9369, instance 9370, instance 9371, instance 9372, instance 9373, instance 9374, instance 10053, instance 10054, instance 10055, instance 10056, instance 10057, instance 10058 , instance 10059, instance 10060, instance 10061, instance 10062, instance 10063, instance 10064, instance 10065, instance 10066, instance 10067, instance 10068, instance 10069, instance 10070, instance 10071, instance 10072, instance 10073, instance 10074, instance 10075, instance 10076, instance 10077, instance 10078, instance 10079, instance 10080, instance 10081, instance 10082, instance 10083, instance 10084, instance 10085, instance 10086, instance 10087, instance 10088, instance 10089, instance 10090, instance 10091, instance 10092, instance 10093, instance 10094, instance 10095, Example 10096, instance 10097, instance 10098, instance 10099, instance 10100, instance 10102, instance 10103, instance 10104, instance 10105, instance 10106, instance 10107, instance 10108, instance 10109, instance 10110, instance 10111, instance 10112, instance 10113 Example 10114, instance 10115, instance 10116, instance 10117, instance 10118, instance 10119, instance 10120, instance 10121, instance 10148, instance 10149, instance 10150, instance 10151, instance 10152, instance 10153, instance 10154, instance 10155, instance 10156, instance 10157, instance 10158, instance 10159, instance 10160, instance 10161, instance 10162, instance 10163, instance 10164, instance 10165, instance 10166, instance 10167, instance 10168, instance 10169, instance 10170, instance 10171, instance 10172, Example 10173, Example 10174, Example 10175, Example 10176, Example 10177 Example 10178, instance 10179, instance 10180, instance 10181, instance 10182, instance 10183, instance 10184, instance 10185, instance 10186, instance 10187, instance 10188, instance 10189, instance 10190, instance 10191, instance 10192, instance 10193, instance 10194 Example 10195, instance 10533, instance 10535, instance 10542, instance 10543, instance 10551, instance 10640, instance 10556, instance 10557, instance 10558, instance 10559, instance 10560, instance 10561, instance 10562, instance 10563, instance 10564, instance 10565, example 10566, example 10567, example 10568, example 10569, example 10570, instance 10571, instance 10572, instance 10573, instance 10574, instance 10575, instance 10576, instance 10577, instance 10078, instance 10579, instance 10580, instance 10581, Example 10582, example 10583, example 10584, example 10585, example 10586, example 10587, example 10588, example 10589, example 10590, instance 10591, instance 10592, instance 10593, instance 10594, instance 10595, instance 10596, instance 10597, instance 10598, instance 10599, instance 10600, instance 10601, instance 10602, instance 10603, instance 10604, instance 10605, instance 10606, Example 10607, instance 10608, instance 10609, instance 10611, instance 10610, instance 10612, instance 10613, instance 10614, instance 10615, instance 10616, instance 10617, instance 10618, instance 10619, instance 10620, instance 10621, instance 10622, instance 10623 , instance 10624, instance 10625, instance 10626, instance 10627, instance 10628, instance 10629, instance 10630, instance 10631, instance 10632, instance 10633, instance 10634, instance 10635, instance 10636, example 10637, and example 10638; Acceptable salt. A compound selected from the group consisting of example 1240, instance 1241, instance 1244, instance 1245, instance 1246, instance 1247, instance 1248, instance 1250, instance 1251, instance 1252, instance 1255, instance 1256, instance 1257, instance 1258, instance 1259 Example 1260, instance 1261, instance 1262, instance 1272, instance 1273, instance 1275, instance 1276, instance 1277, instance 1278, instance 1279, instance 1280, instance 1281, instance 1282, instance 1283, instance 1285, instance 1289, instance 1306, instance 1309, instance 1501, instance 1502, instance 1503, instance 1504, instance 1505, instance 1506, instance 1507, instance 1508, instance 1509, instance 1510, instance 1511, instance 1512, instance 1513, instance 1514, instance 1519, Example 1520, Example 1522, Example 1523, Example 1525, Example 1526, Example 1528, Example 1529, Example 1530, Example 1531, Example 1532, Example 1533, Example 1534, Example 1535, Example 1536, Example 1537, Example 1538, Example 1539 , instance 1542, instance 5148, instance 5150, instance 5151, instance 5152, instance 5154, instance 5156, instance 5157, instance 5158, instance 5159, instance 5160, instance 5161, instance 5162, instance 5164, instance 5165, instance 5167, instance 5169, instance 5170, instance 5174, instance 5175, instance 5177, instance 5178, instance 5179, Example 5182, instance 5183, instance 5184, instance 5186, instance 5189, instance 5190, instance 5191, instance 5196, instance 5198, instance 5200, instance 5201, instance 5202, instance 5204, instance 5205, instance 5207, instance 5208, instance 5209 , instance 5210, instance 5211, instance 5214, instance 5215, instance 5216, instance 5218, instance 5219, instance 5221, instance 5223, instance 5224, instance 5225, instance 5228, instance 5229, instance 5230, instance 5232, instance 5233, instance 5234, instance 5236, instance 5237, instance 5240, instance 5242, instance 5243, instance 5245, instance 5246, instance 5247, instance 5248, instance 5249, instance 5251, instance 5252, instance 5255, instance 5256, instance 5257, instance 5260, Example 5261, instance 5262, instance 5263, instance 5264, instance 5265, instance 6168, instance 6262, instance 6272, instance 7067, instance 7068, real 7069, instance 7070, instance 7071, instance 7072, instance 7073, instance 7075, instance 7077, instance 7078, instance 7079, instance 7080, instance 7081, instance 7082, instance 7083, instance 7084, instance 7085, instance 7806, instance 7807, Example 7088, instance 7088, instance 7089, instance 7090, preparation instance 7091, instance 7092, instance 7093, instance 7094, instance 7905, instance 7096, instance 7097, instance 7908, instance 7099, instance 7100, instance 7101, instance 7102, instance 7103, instance 7104, instance 7105, instance 7106, instance 7107, instance 7108, instance 7109, instance 7110, instance 7111, instance 7112, instance 7113, instance 7114, instance 7115, instance 7116, instance 7117, instance 7118, instance 7119, Example 7120, instance 7121, instance 7122, instance 7123, instance 7124, instance 7125, instance 7126, instance 7127, instance 7128, instance 7129, instance 7130, instance 7131, instance 7132, instance 7133, instance 7134, instance 7135, instance 7136, instance 7137, instance 7138, instance 7139, instance 7152, instance 9115, Example 9116, instance 9117, instance 9118, instance 9119, instance 9120, instance 9121, instance 9122, instance 9203, instance 9124, instance 9125, instance 9126, instance 9127, instance 9128, instance 9129, instance 9130, instance 9131, instance 9132 Example 9133, instance 9134, instance 9135, instance 9136, instance 9137, instance 9138, instance 9139, instance 9140, instance 9141, instance 9142, instance 9143, instance 9144, instance 9145, instance 9146, instance 9147, instance 9148, instance 9149, instance 9150, instance 9155, instance 9156, instance 9157, instance 9158, instance 9159, instance 9160, instance 9161, instance 9162, instance 9163, instance 9164, instance 9165, instance 9168, instance 9169, instance 9170, instance 9171 Example 9172, instance 9173, instance 9174, instance 9175, instance 9176, instance 9177, instance 9178, instance 9179, instance 9180, instance 9181, real 9182, instance 9183, instance 9184, instance 9185, instance 9186, instance 9187, instance 10012, instance 10013, instance 10014, instance 10015, 10016, instance 10017, instance 10018, instance 10019, instance 10020, instance 10021, instance 10022, instance 10023, instance 10024, instance 10025, instance 10034, instance 10035, instance 10036, instance 10037, instance 10042, instance 10043, instance 10044, instance 10045, instance 10047, instance 10049, instance 10050, instance 10051, instance 10052, instance 10122, Example 10123, instance 10126, instance 10127, instance 10128, instance 10129, instance 10130, instance 10131, instance 10132, instance 10136, instance 10137, instance 10138, instance 10139, instance 10140, instance 10141, instance 10142, instance 10143, instance 10144 , instance 10145, instance 10146, instance 10147, instance 10196, instance 10197, instance 10198, instance 10199, instance 10200, instance 10201, instance 10202, instance 10203, instance 10203, instance 10204, instance 10205, instance 10206, instance 10207, instance 10208, instance 10209, instance 10210, instance 10211, Instance 10212, instance 10213, instance 10214, instance 10215, instance 10216, instance 10217, instance 10218, instance 10219, instance 10530, instance 10531, instance 10532, instance 10534, instance 10536, instance 10539, instance 10540, instance 10544, instance 10552 Example 10553, Example 10554, Example 10555, Example 11001, Example 11002, Example 11003, and Example 11004; or a pharmaceutically acceptable salt thereof. A compound selected from the group consisting of example 1290, example 1291, instance 1292, instance 1293, instance 1294, instance 1295, instance 1296, instance 1297, instance 1298, instance 1299, instance 1300, instance 1301, instance 1302, instance 1303, instance 1304 , instance 1305, instance 5153, instance 5155, instance 5163, instance 5166, instance 5168, instance 5171, instance 5176, instance 5180, instance 5181, instance 5187, instance 5112, instance 5199, instance 5203, instance 5213, instance 5220, instance 5222, instance 5226, instance 5227, instance 5231, instance 5238, instance 5241, instance 5253, instance 5254, instance 5259, instance 6118, instance 6119, instance 6128, instance 6134, instance 6135, instance 6136, instance 6137, instance 6140, Example 6141, instance 6146, instance 6156, instance 6158, instance 6162, instance 6170, instance 6172, instance 6176, instance 6180, instance 6183, instance 6184, instance 6185, instance 6189, instance 6201, instance 6202, instance 6203, instance 6204 , instance 6207, instance 6209, instance 6213, instance 6216, instance 6224, instance 6228, instance 6236, instance 6244, instance 6247, instance 6249 6250 examples, examples 6258, instance 6266, instance 6267, instance 6269, instance 6274, instance 6276, instance 6283, instance 6284, instance 6288, instance 6289, instance 6293, instance 6296, instance 6297, instance 6301, instance 6303, instance 6309, instance 6310, Example 6324, instance 6131, instance 6334, instance 6335, instance 6338, instance 6344, instance 6347, instance 6350, instance 6631, instance 6363, instance 6365, instance 6369, instance 6370, instance 6372, instance 6380, instance 6383, instance 6384 , instance 6389, instance 6390, instance 6391, instance 6392, instance 6393, instance 6394, instance 6395, instance 6396, instance 6397, instance 6398, instance 6399, instance 6400, instance 6401, instance 6402, instance 6403, instance 6404, instance 6405, instance 6406, instance 6407, instance 6408, instance 6409, instance 6410, instance 6411, instance 6412, instance 6415, instance 6416, instance 7140, instance 7141, instance 7142, instance 7143, instance 7144, instance 7145, instance 7146, Example 7147, Example 7148, Example 7151, Example 7153, and Example 7154. A use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for enhancing, stimulating and/or enhancing an immune response. The use of claim 4, wherein the medicament further comprises another medicament, or is administered before, after or at the same time as the other medicament. The use of claim 5, wherein the other agent is an antimicrobial agent, an antiviral agent, a cytotoxic agent, and/or an immune response modifier. A use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for inhibiting the growth, proliferation and/or metastasis of cancer cells. The use of claim 7, wherein the cancer is selected from the group consisting of melanoma, renal cell carcinoma, squamous non-small cell lung cancer (NSCLC), non-squamous NSCLC, colorectal cancer, castration-refractory prostate cancer, ovarian cancer, gastric cancer, Hepatocellular carcinoma, pancreatic cancer, head and neck scales Cellular cancer, esophageal cancer, gastrointestinal cancer and breast cancer, and hematological malignancies. A use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for the treatment of an infectious disease. The use of claim 9, wherein the infectious disease is caused by a virus. The use of claim 10, wherein the virus is selected from the group consisting of HIV, hepatitis A, hepatitis B, hepatitis C, herpes virus, and influenza. A use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for the treatment of septic shock. Use of a compound according to any one of claims 1 to 3 for the manufacture of a medicament for blocking the interaction of PD-L1 with PD-1 and/or CD80.