Peptide amide compounds, methods of making and using the same

Abstract

The application discloses a peptide amide compound or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt or a co-crystal thereof, which has a structural general formula (I) shown in the figure: the peptide amide compound has the advantages of better activity, smaller side effect and better drug property as a K opioid receptor agonist.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field and relates to a peptide amide compound, its preparation method, and its application. Specifically, it relates to the compound of this invention and its application in the preparation of drugs for the prevention and / or treatment of diseases mediated by K-opioid receptor (KOR receptor) agonists. Technical Background

[0002] Opioid receptors can be mainly classified into four types: classical μ, δ, K, and atypical noxious peptide (NOP) receptors. μ, δ, and K receptors each have different subtypes. K-opioid receptor agonists have shown analgesic effects, weak respiratory depression, and low addictive potential. Furthermore, K-opioid receptor agonists also possess anti-inflammatory activity and can be used to prevent and treat cardiovascular diseases, pruritus caused by uremia and other kidney diseases, irritable bowel syndrome, intestinal obstruction, and other gastrointestinal diseases. The development of selective K-opioid receptor agonists shows promising potential. K-opioid receptors have multiple subtypes. Besides the classical K1, there are also K2 and K3 subtypes. They are distributed in the human brain and placental tissue. K-opioid receptors participate in analgesia and are related to neuroendocrine and immune regulation. Endopioid peptidergic neurons → release endopioid peptides (enkephalins) → stimulate opioid receptors → through G protein coupling mechanism → inhibit AC → Ca influx decreases, K efflux increases → presynaptic neurotransmitter (substance P, etc.) release decreases → postsynaptic membrane hyperpolarization → prevents the conduction and transmission of pain impulses → analgesia.

[0003] In primates, opioids have three receptors: μ (MOR), K (KOR), and δ (DOR). Opioids (heroin, morphine, codeine) can induce itching. Systemic administration of μ receptor agonists induces scratching, but K and δ receptor agonists inhibit scratching. This phenomenon is not mediated by histamine. Conversely, K receptor agonists, similar to μ receptor antagonists, can suppress opioid-induced itching by preventing or antagonizing itching. These agonists have been developed and applied for purposes including treating heroin and alcohol dependence, combating post-anesthesia depression, anesthetic overdose, and opioid detoxification.

[0004] Physiological opioids (enkephalin, endorphins, and dynorphins) are involved in the pathophysiological mechanisms of pruritus, particularly in atopic dermatitis (AD), uremic pruritus, and hepatic pruritus. The main peptides are β-endorphin (MOR agonist) and dynorphin A (KOR agonist). They function at the central, spinal, and peripheral levels. Nonhistamine-dependent opioid-dependent pruritus appears to originate in unmyelinated C fibers of the epidermis. Behavioral experiments explained a significant reduction in scratching behavior in wild-type KOR and MOR gene knockout mice in a xerotic dermatitis model. These results reveal that MOR and KOR are crucial for cutaneous homeostasis, epidermal nerve fiber regulation, and the pathophysiology of pruritus. In AD patients, only the K receptor system is downregulated, while the μ receptor system is absent. Further studies found that patients treated with psoralen and ultraviolet light showed some recovery of the K receptor system, accompanied by a decrease in pruritus visual analog scale scores. These results suggest that intradermal opioids are closely related to the regulation of pruritus in AD patients. These new findings contribute to our understanding of the mechanisms controlling peripheral pruritus.

[0005] Patent (WO2013184794) reports a novel peptide-based K-opioid receptor agonist, CR-845. This tetrapeptide, containing D-configured amino acids, has demonstrated strong long-lasting analgesic activity and low addictive potential in clinical trials. CR-845 has a broad range of indications, with initiated clinical trials including acute pain, uremic pruritus, postoperative abdominal pain, osteoarthritis, hip and musculoskeletal disorders, rheumatic diseases, postoperative pain, pruritus, and chronic kidney disease. However, Phase III clinical trials of CR-845 have shown that it can cause side effects such as hypernatremia. Therefore, obtaining novel K-opioid receptor agonists with better activity, fewer side effects, and better drug-like properties remains attractive. This invention provides a novel K-opioid receptor (KOR receptor) agonist compound that surprisingly exhibits excellent effects and functions. Summary of the Invention

[0006] The purpose of this invention is to provide a novel K opioid receptor agonist with a novel structure, better biological activity, fewer side effects, and better drug-like properties, as well as its preparation method and its use in medicine.

[0007] To address the above problems, the present invention provides a peptide amide compound or its stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, or cocrystal represented by general structural formula (Ⅰ), having the following general structural formula:

[0008]

[0009] in,

[0010] R aSelected from H, C 1-6 alkyl;

[0011] R b Selected from

[0012] R c Selected from C 1-6 alkyl;

[0013] R d R e Each is independently selected from H and C. 1-6 Alkyl, C 1-6 Alkyl-NH2, 3- to 6-membered cycloalkyl, C 1-6 Alkyl-OC 1-6 Alkyl-OC 1-6 Alkyl group, -S (=O) p -C 1-6 Alkyl group, -C(=O)C 1-6 alkyl;

[0014] R f Selected from

[0015] R h Selected from phenyl, H;

[0016] R g Selected from H, -OH, -COOH, -COOC 1-6 Alkyl, -CONHC 1-6 alkyl;

[0017] Or R a and R b Together with the N atom it is attached to, it forms the following structure:

[0018]

[0019] R1 is selected from -CH2C(=O)R j -CH2-OC 1-6 Alkyl group, -CH2NH(C=O)C 1-10 Alkyl, H, -NH2, -NH(C=S)NH-C 1-6 Alkyl group, -NH(C=NH)NH-C 1-6 Alkyl group, -NH(C=O)NH2, -NH(C=O)NH-C 1-6 Alkyl group, -NH(C=O)(C=O)NH-C 1-6 Alkyl, -CN, -C(=O)NH2, -CH2OH, -NR7(C=O)R8,

[0020] R2 is selected from H, -CH2NR k R L Halogen, deuterium, -C 1-6 Alkyl, -CN, -C 1-6 Alkyl groups -NH2, -(CH2) q -COOH, -CONH2, -C(=O)C(=O)NH-C 1-6 Alkyl group, -(CH2) q -C(=O)NHR 12 ,

[0021] R3 is selected from -C(=O)OC 1-6 alkyl,

[0022] R4 is selected from H and C. 1-6 Alkyl, -C(=O)-C 1-6 alkyl;

[0023] The R j Selected from H, -OH, -OC 1-6 Alkyl, -NR x R y C 1-6 alkyl;

[0024] The R k and R L Each independently selected from H and C 1-6 Alkyl, 3-7 membered cycloalkyl, -S (=O) p -C 1-6 Alkyl, -C(=O)R z -C(=O)OR z -C(=O)NHR z -C(=S)NHR z -C(=NH)NHRz;

[0025] R5 is selected from -C(=O)C(=O)OH, -C(=O)C(=O)OCH2CH3, -C(=O)-C 1-6 alkyl;

[0026] R6 is selected from

[0027] R7 is selected from H and C. 1-6 Alkyl, 3- to 6-membered cycloalkyl; the C 1-6 The alkyl group may be further replaced by 1 to 3 deuteriums, 1 to 3 halogens, or 3 to 6-membered cycloalkyl groups;

[0028] R8 is selected from -C 1-6 Alkyl, -OC 1-6 Alkyl, -OCH2CF3, -COOH

[0029]

[0030] R9 is selected from halogens, hydrogen (H), deuterium, and carbon. 1-6 alkyl;

[0031] R 10 R 11 Each is independently selected from H, deuterium, and halogens;

[0032] R 12 Selected from -C 1-6 alkyl,

[0033] R is selected from halogen, -C 1-6 Alkyl, -OC 1-6 Alkyl, wherein the C 1-6 The alkyl group can be further replaced by 1 to 3 deuterium or halogen groups;

[0034] The R x R y and R z Each is independently selected from H and -C 1-6 Alkyl, 3-7 membered cycloalkyl, phenyl, -C 1-6 Alkyl-phenyl, -OC 1-6 Alkyl, -C 1-6 Alkyl-OC 1-6 Alkyl-OC 1-6 Alkyl-OC 1-6 Alkyl, OH; wherein the alkyl or cycloalkyl group may optionally be replaced by 1, 2, or 3 deuterium, halogen, or NH2;

[0035] n is selected from 0, 1, 2, 3, and 4;

[0036] m is selected from 0, 1, 2, and 3;

[0037] q is selected from 0, 1, 2, and 3;

[0038] p is selected from 1 and 2.

[0039] In some embodiments, the R a and R b Together with the N atom it is attached to, it forms the following structure:

[0040]

[0041] In some embodiments, the R a and R b Together with the N atom it is attached to, it forms the following structure:

[0042]

[0043] in,

[0044] R1 is selected from H and NH2;

[0045] R2 is selected from -(CH2). q -C(=O)NHR 12 ;

[0046] q is selected from 0, 1, 2, and 3.

[0047] In some embodiments, the peptide amide compound or its stereoisomers, hydrates, metabolites, solvates, pharmaceutically acceptable salts or cocrystals have the following general structural formula (II):

[0048]

[0049] in,

[0050] R1 is selected from -NR7(C=O)R8;

[0051] R2 is selected from H, -C 1-6 alkyl.

[0052] In some preferred embodiments, R1 is selected from -NR7(C=O)R8;

[0053] R2 is selected from H;

[0054] R7 is selected from H, -C 1-6 alkyl.

[0055] In some preferred embodiments, R8 is selected from -OC 1-6 alkyl,

[0056] In some embodiments, the peptide amide compound or its stereoisomers, hydrates, metabolites, solvates, pharmaceutically acceptable salts or cocrystals have the following general structural formula (III):

[0057]

[0058] On the other hand, the present invention provides a pharmaceutical composition comprising the compound shown above, or its stereoisomers, hydrates, metabolites, solvates, pharmaceutically acceptable salts or cocrystals, and one or more pharmaceutically acceptable carriers and / or excipients.

[0059] On the other hand, the present invention provides the use of the above-mentioned compound or its stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt or cocrystal, or the above-mentioned pharmaceutical composition for the manufacture of a medicament for the treatment or prevention of diseases or conditions related to K opioid receptors in mammals.

[0060] Preferably, the diseases associated with the K opioid receptor are selected from pain, inflammation, itching, edema, hyponatremia, hypokalemia, intestinal obstruction, cough, and glaucoma.

[0061] Furthermore, the preferred pain categories are neuropathic pain, trunk pain, visceral pain, skin pain, arthritis pain, kidney stone pain, uterine spasms, dysmenorrhea, endometriosis, indigestion, postoperative pain, post-medical treatment pain, eye pain, ear pain, fulminant cancer pain, and pain associated with GI disorders.

[0062] The pruritus associated with the K opioid receptor (KOR receptor) involved in this invention can be any pruritus disease and condition, such as eye pruritus, pruritus associated with end-stage renal disease (many of whom undergo kidney dialysis) and other forms of cholestasis, skin conditions, insect bite pruritus, and drug-induced pruritus.

[0063] Among them, itchy eyes, such as itchy eyes related to conjunctivitis;

[0064] Other forms of cholestasis include primary biliary cirrhosis, intrahepatic cholestasis of pregnancy, chronic cholesterol liver disease, uremia, malignant cholestasis, and jaundice;

[0065] Skin conditions such as eczema (dermatitis), tinea, polycythemia, lichen planus, chronic simple lichen planus, lice, thyrotoxicosis, tinea pedis, urticaria, scabies, vaginitis, and anal itching associated with hemorrhoids; among which, eczema (dermatitis) includes atopic dermatitis or contact dermatitis. Specific Implementation

[0066] The following embodiments illustrate the technical solution of the present invention in detail, but the scope of protection of the present invention includes, but is not limited to, these embodiments.

[0067] Example 1

[0068] Synthesis of (R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)-N-methyl-N-((S)-1-phenyl-2-(pyrrolidone-1-yl)ethyl)hexamethylene trihydrochloride (1)

[0069]

[0070] Step 1: Synthesis of compounds 1-3

[0071] Dissolve 1-1 (350 mg, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) in tetrahydrofuran (10 mL). Stir the reaction mixture at room temperature for 30 min. Then add 1-2 (104 mg, 0.51 mmol) and stir the mixture overnight at room temperature. After the reaction is complete as monitored by TLC, wash the reaction mixture once with 1N Na₂CO₃ and once with 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using eluent V. 二氯甲烷 :V 甲醇 =From 100:1 to 40:1, a white solid 1-3 (162 mg, yield: 37.1%), [M+H) was obtained. + :940.3.

[0072] The solid-phase synthesis of similar peptides used in section 1-1 was also performed using this method, and will not be described in detail elsewhere. The synthesis procedure is as follows: Weigh 10g of 2-Cl-resin into a peptide synthesis tube, add 50mL of dichloromethane, puff with nitrogen, add SM-1 (Fmoc-D-Lys(Boc)-OH, 4.68g, 10mmol) and DIEA (2.58g, 20mmol), puff with nitrogen for 1h, add methanol (5mL), puff with nitrogen for 30min, dry under vacuum, wash 3 times with DMF (50mL*3), dry under vacuum, and use directly for the next step; add 50mL of piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, puff with nitrogen for 30min, dry under vacuum, wash 3 times with DMF (50mL*3), dry under vacuum, and use directly for the next step; add 50mL of piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, puff with nitrogen for 30min, dry under vacuum, wash 3 times with DMF (50mL*3), dry under vacuum, and use directly for the next step; add 50mL of piperidine / DMF (v:v = 1:4) to the peptide synthesis tube. DMF, nitrogen agitation, add Fmoc-D-Leu-OH (7.06 g, 20 mmol), HATU (7.6 g, 20 mmol), DIEA (2.58 g, 20 mmol), nitrogen agitation for 1 h, drain the reaction solution, wash 3 times with DMF (50 mL * 3), drain directly for the next step; add 50 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, agitate with nitrogen for 30 min, drain the liquid, wash 3 times with DMF (50 mL * 3), drain directly for the next step; add 50 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, agitate with nitrogen for 30 min, drain the liquid, wash 3 times with DMF (50 mL * 3), drain directly for the next step; add 50 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube. DMF, nitrogen agitation, add Fmoc-D-Phe-OH (7.74 g, 20 mmol), HATU (7.6 g, 20 mmol), DIEA (2.58 g, 20 mmol), nitrogen agitation for 1 h, drain the reaction solution, wash 3 times with DMF (50 mL * 3), drain directly for the next step; add 50 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, agitate with nitrogen for 30 min, drain the liquid, wash 3 times with DMF (50 mL * 3), drain directly for the next step; add 50 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, agitate with nitrogen for 30 min, drain the liquid, wash 3 times with DMF (50 mL * 3), drain directly for the next step; add 50 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube. DMF was added under nitrogen agitation, followed by the addition of Boc-D-Phe-OH (5.38 g, 20 mmol), HATU (7.6 g, 20 mmol), and DIEA (2.58 g, 20 mmol). The mixture was agitated under nitrogen for 1 hour, then the reaction solution was dried. The solution was washed three times with DMF (50 mL x 3), dried, and then washed three times with MeOH (50 mL x 3). The solution was dried again, followed by the addition of DCM (50 mL x 3), and then dried for use in the next step. 50 mL of hexafluoroisopropanol / DCM (v:v = 1:4) was added to the peptide synthesis tube, and the mixture was agitated under nitrogen for 30 minutes. The solution was dried, and this process was repeated three times. The resulting liquids were combined and evaporated to dryness to obtain a white solid 1-1 (7.01 g, yield: 93.0%), [M+H]. + :754.4.

[0073] Step 2: Synthesis of Compound 1

[0074] Compounds 1-3 (162 mg, 0.17 mmol) were dissolved in ethyl acetate (5 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 h. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 1 was a white solid (102 mg, yield: 69.7%), [M+H]. + :740.2.

[0075] 1 H NMR (400MHz, CD3OD), δ7.47-7.33(m,10H),6.31(m,1H),4.77(m,1H),4.63(m,1H),4.46(m,1H),4.27-4.15(m,1H),3.8 5-3.77(m,3H),3.38-3.24(m,4H),3.05-2.88(m,8H),2.18-2.08(m,5H),1.79-1.51(m,13H),1.02(d,3H),0.99(d,3H).

[0076] Example 2

[0077] Synthesis of (R)-N-((R)-6-amino-1-(4-(2-(3,4-dichlorophenyl)acetyl)piperazin-1-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide dihydrochloride (2)

[0078] Step 1: Synthesis of compounds 2-3

[0079] 2-2 (0.88 g, 4.32 mmol), HATU (1.89 g, 4.97 mmol), and DIEA (1.11 g, 8.64 mmol) were dissolved in tetrahydrofuran (15 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 2-1 (0.7 g, 3.76 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of 100:1 to 40:1 yielded a white solid 2-2 (1.15 g, yield: 81.9%), [M+H]. + :373.2.

[0080] Step 2: Synthesis of compounds 2-4

[0081] Compounds 2-3 (1.15 g, 3.08 mmol) were dissolved in ethyl acetate (20 mL), and dioxane hydrochloride (4 M, 10 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. Target compounds 2-4 were white solids (0.92 g, yield: 96.4%), [M+H]. + :273.1.

[0082] Step 3: Synthesis of compounds 2-5

[0083] Dissolve 1-1 (0.35 g, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) in tetrahydrofuran (10 mL). Stir the reaction mixture at room temperature for 30 min. Then add 2-4 (158 mg, 0.51 mmol) and stir the mixture overnight at room temperature. After the reaction is complete as monitored by TLC, wash the reaction mixture once with 1N Na₂CO₃ and once with 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using eluent V. 二氯甲烷 :V 甲醇 =From 100:1 to 35:1, a white solid 2-5 (240 mg, yield: 51.2%) was obtained, [M+H]. + :1008.5.

[0084] Step 4: Synthesis of Compound 2

[0085] Compound 2-5 (240 mg, 0.24 mmol) was dissolved in ethyl acetate (6 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 2 was a white solid (152 mg, yield: 72.5%), [M+H]. + :808.4.

[0086] 1 H NMR (400MHz, CD3OD), δ7.49-7.20(m,13H),4.84(m,1H),4.73(m,1H),4.41(m,1H),4.14(m,1H),3.84-3.71(m ,4H),3.62-3.45(m,4H),3.36-3.21(m,3H),3.04-2.89(m,5H),1.80-1.42(m,9H),1.00(d,3H),0.95(d,3H).

[0087] Example 3

[0088] Synthesis of 4-amino-1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-(4-(trifluoromethoxy)phenyl)propamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)piperidine-4-carboxylic acid trihydrochloride (3)

[0089]

[0090] Step 1: Synthesis of compound 3-2

[0091] The solid-phase synthesis used in 3-2 is performed as follows:

[0092] Weigh 1 g of 2-Cl-resin into a peptide synthesis tube, add 5 mL of dichloromethane, puff under nitrogen, add 3-1 (0.47 g, 1 mmol) and DIEA (0.26 g, 2 mmol), puff under nitrogen for 1 h, add 1 mL of methanol, puff under nitrogen for 30 min, dry under vacuum, wash 3 times with DMF (5 mL * 3), dry under vacuum, and use directly for the next step; add 5 mL of piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, puff under nitrogen for 30 min, dry under vacuum, wash 3 times with DMF (5 mL * 3), dry under vacuum, and use directly for the next step; add 5 mL of piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, puff under nitrogen for 30 min, dry under vacuum, wash 3 times with DMF (5 mL * 3), dry under vacuum, and use directly for the next step; add 5 mL of piperidine / DMF (v:v = 1:4) to the peptide synthesis tube. DMF, nitrogen agitation, add Fmoc-D-Lys(Boc)-OH (0.94 g, 2 mmol), HATU (0.76 g, 2 mmol), DIEA (0.26 g, 2 mmol), nitrogen agitation for 1 h, drain the reaction solution, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, agitate with nitrogen for 30 min, drain the liquid, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, agitate with nitrogen for 30 min, drain the liquid, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube. DMF, nitrogen agitation, add Fmoc-D-Leu-OH (0.71 g, 2 mmol), HATU (0.76 g, 2 mmol), DIEA (0.26 g, 2 mmol), nitrogen agitation for 1 h, drain the reaction solution, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, agitate with nitrogen for 30 min, drain the liquid, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube. DMF, nitrogen purging, add Fmoc-D-Phe-OH (0.77 g, 2 mmol), HATU (0.76 g, 2 mmol), DIEA (0.26 g, 2 mmol), nitrogen purging for 1 h, drain the reaction solution, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, nitrogen purging for 30 min, drain the liquid, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 50 mL DMF to the peptide synthesis tube, nitrogen purging, add (R)-2-(tert-butoxycarbonyl)amino)-3-(4-(trifluoromethoxy)phenyl)propionic acid (0.70 g, 2 mmol), HATU (0.76 g, 2 mmol), DIEA (0.26 g, 2 mmol), nitrogen purging for 1 h, drain the reaction solution, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add (R)-2-(tert-butoxycarbonyl)amino)-3-(4-(trifluoromethoxy)phenyl)propionic acid (0.70 g, 2 mmol), HATU (0.76 g, 2 mmol), DIEA (0.26 g, 2 mmol), nitrogen purging for 1 h, add Fmoc-D-Phe-OH (0.77 g, 2 mmol), HATU (0.76 g, 2 mmol), DIEA (0.26 g, 2 mmol), nitrogen purging for 1 h, ...26 g (2 mmol), agitated under nitrogen for 1 h, the reaction solution was dried under vacuum, washed 3 times with DMF (5 mL * 3), dried under vacuum, washed 3 times with MeOH (5 mL * 3), dried under vacuum, washed 3 times with DCM (5 mL * 3), dried under vacuum and used directly for the next step; 5 mL of hexafluoroisopropanol / DCM (v:v = 1:4) was added to the polypeptide synthesis tube, agitated under nitrogen for 30 min, the liquid was dried under vacuum, and the process was repeated 3 times. The resulting liquids were combined and evaporated to dryness to obtain 3-2 as a white solid (0.58 g, yield: 54.3%), [M + H). + :1064.5.

[0093] Step 2: Synthesis of Compound 3

[0094] Compound 3-2 (580 mg, 0.55 mmol) was dissolved in ethyl acetate (12 mL), and dioxane hydrochloride (4 M, 6 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 3 was a white solid (310 mg, yield: 65.1%), [M+H]. + :764.4.

[0095] 1 H NMR(400MHz,CD3OD),δ7.47-7.22(m,9H),4.73(m,1H),4.41(m,1H),4.19(m,2H),3.88(m,1H),3 .74(m,1H),3.34-3.23(m,6H),3.02-2.83(m,4H),2.27-1.46(m,13H),1.00(d,3H),0.96(d,3H).

[0096] Example 4

[0097] Synthesis of (R)-N-((R)-6-amino-1-(4-((E)-3-(furan-3-yl)acryloyl)piperazin-1-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide dihydrochloride (4)

[0098] Step 1: Synthesis of compound 4-2

[0099] 4-1 (0.64 g, 4.63 mmol), HATU (2.01 g, 5.29 mmol), and DIEA (1.14 g, 8.82 mmol) were dissolved in tetrahydrofuran (15 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 2-1 (0.82 g, 4.41 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of 100:1 to 40:1 yielded a white solid 4-2 (0.95 g, yield: 70.4%), [M+H]. + :307.2.

[0100] Step 2: Synthesis of Compound 4-3

[0101] Compound 4-2 (0.95 g, 3.10 mmol) was dissolved in ethyl acetate (20 mL), and dioxane hydrochloride (4 M, 10 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 4-3 was a white solid (0.75 g, yield: 99.6%), [M+H]. + :207.2.

[0102] Step 3: Synthesis of compounds 4-5

[0103] Dissolve 1-1 (0.35 g, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) in tetrahydrofuran (10 mL). Stir the reaction mixture at room temperature for 30 min. Then add 4-3 (124 mg, 0.51 mmol) and stir the mixture overnight at room temperature. After the reaction is complete as monitored by TLC, wash the reaction mixture once with 1N Na₂CO₃ and once with 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 =From 100:1 to 35:1, a white solid 4-5 (272 mg, yield: 62.1%) was obtained, [M+H]. + :942.5.

[0104] Step 4: Synthesis of Compound 4

[0105] Compound 4-5 (272 mg, 0.29 mmol) was dissolved in ethyl acetate (6 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 4 was a white solid (192 mg, yield: 81.6%), [M+H]. + :742.4.

[0106] 1 H NMR(400MHz, CD3OD), δ7.84(m,1H),7.58-53(m,2H),7.39-7.22(m,10H),6.91-6.82(m,2H),4.72(m,1H),4.43-4.39(m,1H),4.15-4. 11(m,1H),3.88-3.83(m,4H),3.65-3.60(m,4H),3.35-3.22(m,3H),3.01-2.90(m,4H),1.85-1.32(m,9H),1.00(d,3H),0.96(d,3H).

[0107] Example 5

[0108] Synthesis of (R)-2-amino-N-((R)-1-((R)-1-((R)-6-amino-1-(4-((E)-3-(furan-3-yl)acryloyl)piperazin-1-yl)-1-oxohex-2-yl)amino)-3-cyclopropyl-1-oxopropyl-2-yl)amino)-1-oxo-3-phenylprop-2-yl)-3-phenylpropionamide dihydrochloride (5)

[0109]

[0110] Step 1: Synthesis of Compound 5-3

[0111] 5-1 (350 mg, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) were dissolved in tetrahydrofuran (10 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 4-3 (124 mg, 0.51 mmol) was added, and the mixture was stirred overnight at room temperature. After the reaction was complete as monitored by TLC, the reaction mixture was washed once with 1N Na₂CO₃ and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 =From 100:1 to 35:1, a white solid 5-3 (252 mg, yield: 57.5%) was obtained, [M+H]. +:940.5.

[0112] The solid-phase synthesis used in 5-1 is similar to that in 1-1, and the synthesis procedure is as follows: Weigh 1g of 2-Cl-resin into a peptide synthesis tube, add 5mL of dichloromethane, puff with nitrogen, add (Fmoc-D-Lys(Boc)-OH, 0.47g, 1mmol), DIEA (0.26g, 2mmol), puff with nitrogen for 1h, add methanol (1mL), puff with nitrogen for 30min, dry under vacuum, wash 3 times with DMF (5mL*3), dry under vacuum, and use directly for the next step; add 5mL of piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, puff with nitrogen for 30min, dry under vacuum, wash 3 times with DMF (5mL*3), dry under vacuum, and use directly for the next step. Step 1: Add 5 mL of DMF to the peptide synthesis tube, puff with nitrogen, add N-fluorenemethoxycarbonyl-D-cyclopropylalanine (0.70 g, 2 mmol), HATU (0.76 g, 2 mmol), and DIEA (0.26 g, 2 mmol), puff with nitrogen for 1 h, drain the reaction solution, wash 3 times with DMF (5 mL * 3), drain directly for the next step; Add 5 mL of piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, puff with nitrogen for 30 min, drain the liquid, wash 3 times with DMF (5 mL * 3), drain directly for the next step; Add 5 mL of piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, puff with nitrogen for 30 min, drain the liquid, wash 3 times with DMF (5 mL * 3), drain directly for the next step; Add 5 mL of piperidine / DMF (v:v = 1:4) to the peptide synthesis tube. DMF, nitrogen purging, add Fmoc-D-Phe-OH (0.77 g, 2 mmol), HATU (0.76 g, 2 mmol), DIEA (0.26 g, 2 mmol), nitrogen purging for 1 h, drain the reaction solution, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, nitrogen purging for 30 min, drain the liquid, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 50 mL DMF was added under nitrogen agitation. Boc-D-Phe-OH (0.53 g, 2 mmol), HATU (0.76 g, 2 mmol), and DIEA (0.26 g, 2 mmol) were added and agitated under nitrogen for 1 h. The reaction mixture was then dried under vacuum. The solution was washed three times with DMF (5 mL x 3), dried under vacuum, washed three times with MeOH (5 mL x 3), dried under vacuum, and then washed three times with DCM (5 mL x 3). This solution was used directly in the next step. 5 mL of hexafluoroisopropanol / DCM (v:v = 1:4) was added to the peptide synthesis tube and agitated under nitrogen for 30 min. The liquid was dried under vacuum. This process was repeated three times. The resulting liquids were combined and evaporated to dryness to obtain a white solid (0.65 g, yield: 54.3%), [M+H]. + :752.4.

[0113] Step 2: Synthesis of Compound 5

[0114] Compound 5-3 (252 mg, 0.29 mmol) was dissolved in ethyl acetate (6 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 5 was a white solid (182 mg, yield: 83.5%), [M+H]. + :740.4.

[0115] 1 H NMR(400MHz,CD3OD),δ7.85(m,1H),7.58-53(m,2H),7.38-7.22(m,10H),6.91-6.82(m,2H),4.73(m,1H),4.43-4.39(m,1H),4.18-4.13(m,1H) ,3.86-3.83(m,4H),3.65-3.57(m,4H),3.39-3.22(m,3H),3.01-2.90( m,4H),1.85-1.28(m,9H),0.82(m,1H),0.52-0.45(m,2H),0.18(m,2H).

[0116] Example 6

[0117] Synthesis of (R)-N-((R)-6-amino-1-(4-((E)-3-(furan-3-yl)-N-methacrylamido)piperidin-1-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropionamido)-3-phenylpropionamido)-4-methylpentanamide dihydrochloride (6)

[0118] Step 1: Synthesis of Compound 6-2

[0119] 4-1 (0.47 g, 3.39 mmol), HATU (1.55 g, 4.07 mmol), and DIEA (0.87 g, 6.78 mmol) were dissolved in tetrahydrofuran (12 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 6-1 (0.8 g, 3.73 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of 100:1 to 50:1 yielded a white solid 6-2 (0.91 g, yield: 72.9%), [M+H]. + :335.3.

[0120] Step 2: Synthesis of Compound 6-3

[0121] Compound 6-2 (0.91 g, 2.72 mmol) was dissolved in ethyl acetate (12 mL), and dioxane hydrochloride (4 M, 6 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 6-3 was a white solid (0.71 g, yield: 96.6%), [M+H]. + :235.3.

[0122] Step 3: Synthesis of Compounds 6-5

[0123] Dissolve 1-1 (350 mg, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) in tetrahydrofuran (10 mL). Stir the reaction mixture at room temperature for 30 min. Then add 6-3 (138 mg, 0.51 mmol) and stir the mixture overnight at room temperature. After the reaction is complete as monitored by TLC, wash the reaction mixture once with 1N Na₂CO₃ and once with 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 =100:1 to 35:1 yielded a white solid 6-5 (272 mg, yield: 60.3%), [M+H] + :970.5.

[0124] Step 4: Synthesis of Compound 6

[0125] Compound 6-5 (272 mg, 0.28 mmol) was dissolved in ethyl acetate (6 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 6 was a white solid (192 mg, yield: 81.2%), [M+H]. + :770.4.

[0126] 1H NMR(400MHz,CD3OD),δ7.85(m,1H),7.57-7.52(m,2H),7.36-7.22(m,10H),7.02-6.79(m,2H),4.75-4I.64(m,2H),4.45 -4.38(m,1H),4.18-4.11(m,2H),3.37-3.21(m,3H),3.09-2.93(m,10H),1.87-1.50(m,13H),1.00(d,3H),0.97(m,3H).

[0127] Example 7

[0128] Synthesis of (R)-N-((R)-6-amino-1-(4-((E)-3-(furan-3-yl)acrylamido)piperidin-1-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide dihydrochloride (7)

[0129] Step 1: Synthesis of Compound 7-2

[0130] 4-1 (0.6 g, 4.35 mmol), HATU (1.98 g, 5.22 mmol), and DIEA (1.12 g, 8.70 mmol) were dissolved in tetrahydrofuran (12 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 7-1 (0.79 g, 3.95 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] to 50:1 yielded a white solid 7-2 (0.92 g, yield: 72.8%). + :321.2.

[0131] Step 2: Synthesis of Compound 7-3

[0132] Compound 7-2 (0.92 g, 2.88 mmol) was dissolved in ethyl acetate (12 mL), and dioxane hydrochloride (4 M, 6 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 7-3 was a white solid (0.70 g, yield: 94.9%), [M+H]. + :221.1.

[0133] Step 3: Synthesis of Compounds 7-5

[0134] Dissolve 1-1 (0.35 g, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) in tetrahydrofuran (10 mL). Stir the reaction mixture at room temperature for 30 min. Then add 7-3 (131 mg, 0.51 mmol) and stir the mixture overnight at room temperature. After the reaction is complete as monitored by TLC, wash the reaction mixture once with 1N Na₂CO₃ and once with 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 =100:1 to 35:1 yielded a white solid 7-5 (205 mg, yield: 46.1%), [M+H] + :956.5.

[0135] Step 4: Synthesis of Compound 7

[0136] Compound 7-5 (205 mg, 0.21 mmol) was dissolved in ethyl acetate (6 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 7 was a white solid (115 mg, yield: 64.7%), [M+H]. + :756.4.

[0137] 1 H NMR(400MHz,CD3OD),δ7.82(m,1H),7.55-7.23(m,12H),6.93-6.69(m,1H),6.42-6.33(m,1H),4.75-4.69(m,1H),4.5 1-4.35(m,2H),4.19-3.98(m,3H),3.40-3.21(m,4H),3.05-2.8(m,5H),1.98-1.42(m,13H),1.00(d,3H),0.95(m,3H).

[0138] Example 8

[0139] Synthesis of (R)-2-amino-N-((R)-1-((R)-1-((R)-6-amino-1-oxo-1-(3-oxo-2,8-diazaspiro[4.5]decane-8-yl)hex-2-yl)amino)-3-cyclopropyl-1-oxopropyl-2-yl)amino)-1-oxo-3-phenylprop-2-yl)-3-phenylpropionamide dihydrochloride (8)

[0140] Step 1: Synthesis of Compound 8-3

[0141] 5-1 (350 mg, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) were dissolved in tetrahydrofuran (10 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 8-2 (97 mg, 0.51 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 =100:1 to 35:1 yielded a white solid 8-3 (201 mg, yield: 48.6%), [M+H] + :888.4.

[0142] Step 2: Synthesis of Compound 8

[0143] Compound 8-3 (201 mg, 0.23 mmol) was dissolved in ethyl acetate (6 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 8 was a white solid (139 mg, yield: 80.6%), [M+H]. + :688.4.

[0144] 1 H NMR (400MHz, CD3OD), δ7.33-7.20(m,10H),6.91-6.82(m,2H),4.87(m,1H),4.70(m,1H),4.38(m,1H),4.14(m,1H),3.63-3.51(m ,3H),3.34-3.20(m,3H),3.01-2.92(m,4H),2.40-2.35(m,2H),1.79-1.40(m,13H),0.79(m,1H),0.49-0.44(m,2H),0.16(m,2H).

[0145] Example 9

[0146] Synthesis of N-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)piperidin-4-yl)-N-methylfuran-3-carboxamide dihydrochloride (9)

[0147]

[0148] Step 1: Synthesis of Compound 9-2

[0149] 3-Furfuric acid (0.22 g, 1.95 mmol), HATU (0.89 g, 2.34 mmol), and DIEA (0.50 g, 3.90 mmol) were dissolved in tetrahydrofuran (10 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 6-1 (0.40 g, 1.85 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of 100:1 to 50:1 yielded a white solid 9-2 (0.44 g, yield: 76.4%), [M+H]. + :309.2.

[0150] Step 2: Synthesis of Compound 9-3

[0151] Compound 9-2 (0.44 g, 1.43 mmol) was dissolved in ethyl acetate (8 mL), and dioxane hydrochloride (4 M, 4 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 9-3 was a white solid (0.34 g, yield: 97.3%), [M+H]. + :209.2.

[0152] Step 3: Synthesis of Compound 9-5

[0153] Dissolve 1-1 (350 mg, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) in tetrahydrofuran (10 mL). Stir the reaction mixture at room temperature for 30 min. Then add 9-3 (125 mg, 0.51 mmol) and stir the mixture overnight at room temperature. After the reaction is complete as monitored by TLC, wash the reaction mixture once with 1N Na₂CO₃ and once with 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 =From 100:1 to 40:1, a white solid 9-5 (289 mg, yield: 65.9%) was obtained, [M+H]. + :944.5.

[0154] Step 4: Synthesis of Compound 9

[0155] Compound 9-5 (289 mg, 0.31 mmol) was dissolved in ethyl acetate (6 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 9 was a white solid (209 mg, yield: 83.6%), [M+H]. + :744.4.

[0156] 1 H NMR (400MHz, CD3OD), δ7.93(m,1H),7.61(m,1H),7.39-7.22(m,10H),6.67(m,1H),4.73-4.66(m,2H),4.45-4.38 (m,1H),4.19-4.09(m,2H),3.40-3.21(m,4H),3.05-2.91(m,9H),1.87-1.34(m,13H),1.00(d,3H),0.95(d,3H).

[0157] Example 10

[0158] Synthesis of (R)-N-((R)-6-amino-1-(4-(2-(3,4-dichlorophenyl)-N-methylacetamido)piperidin-1-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide dihydrochloride (10)

[0159] Step 1: Synthesis of Compound 10-2

[0160] 2-2 (0.4 g, 1.95 mmol), HATU (0.89 g, 2.34 mmol), and DIEA (0.50 g, 3.90 mmol) were dissolved in tetrahydrofuran (10 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 6-1 (0.40 g, 1.85 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] was increased from 100:1 to 50:1 to obtain a white solid 10⁻² (0.37 g, yield: 49.4%). + :401.2.

[0161] Step 2: Synthesis of compound 10⁻³

[0162] 10⁻² (0.37 g, 0.92 mmol) was dissolved in ethyl acetate (8 mL), and dioxane hydrochloride (4 M, 4 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 10⁻³ was a white solid (0.31 g, yield: 99.5%), [M+H]. + :301.1. Step 3: Synthesis of compound 10-5

[0163] 1-1 (350 mg, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) were dissolved in tetrahydrofuran (10 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 10-3 (172 mg, 0.51 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] to [M+H] was increased from 100:1 to 40:1 to obtain the target compound as a white solid (256 mg, yield: 53.1%). + :1036.5.

[0164] Step 4: Synthesis of Compound 10

[0165] 10⁻⁵ (256 mg, 0.25 mmol) was dissolved in ethyl acetate (6 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 10 was a white solid (160 mg, yield: 71.3%), [M+H⁺]. + :836.4.

[0166] 1 H NMR (400MHz, CD3OD), δ7.51-7.14(m,13H),4.734.41(m,4H),4.15-4.09(m,2H),3.91-3.75( m,2H),3.42-3.21(m,4H),3.04-2.72(m,8H),1.80-1.39(m,13H),0.99(d,3H),0.95(d,3H).

[0167] Example 11

[0168] Synthesis of (R)-N-((R)-6-amino-1-(4-(3-methylthioureo)piperidin-1-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropionylamino)-3-phenylpropionylamino)-4-methylpentanamide dihydrochloride (11)

[0169]

[0170] Step 1: Synthesis of Compound 11-2

[0171] Methyl isothiocyanate (0.22 g, 3 mmol) was dissolved in dichloromethane (6 mL), and then a solution of 7-1 (0.5 g, 2.5 mmol) in dichloromethane (4 mL) was added to the above solution. The reaction mixture was stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] to

[30] was adjusted from 50:1 to 30:1 to obtain the target compound 11-2 as a white solid (0.62 g, yield: 90.8%). + :274.2.

[0172] Step 2: Synthesis of compound 11-3

[0173] Compound 11-2 (0.62 g, 2.27 mmol) was dissolved in ethyl acetate (12 mL), and dioxane hydrochloride (4 M, 6 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 11-3 was a white solid (0.46 g, yield: 96.7%), [M+H]. + :174.2. Step 3: Synthesis of compound 11-5

[0174] Dissolve 1-1 (350 mg, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) in tetrahydrofuran (10 mL). Stir the reaction mixture at room temperature for 30 min. Then add 11-3 (107 mg, 0.51 mmol) and stir the mixture overnight at room temperature. After the reaction is complete as monitored by TLC, wash the reaction mixture once with 1N Na₂CO₃ and once with 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using eluent V. 二氯甲烷 :V 甲醇The ratio of [M+H] was increased from 80:1 to 25:1 to obtain the target compound 11-5 as a white solid (210 mg, yield: 51.2%). + :909.5.

[0175] Step 4: Synthesis of Compound 11

[0176] Compound 11-5 (210 mg, 0.23 mmol) was dissolved in ethyl acetate (6 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 11 was a white solid (131 mg, yield: 72.6%), [M+H]. + :909.5.

[0177] 1 H NMR(400MHz,CD3OD),δ7.37-7.22(m,10H),4.73(m,1H),4.44-4.40(m,2H),4.17(m,1H),3 .80(m,1H),3.37-3.23(m,5H),2.76(m,2H),1.92-1.41(m,17H),1.00(d,3H),0.95(d,3H).

[0178] Example 12

[0179] Synthesis of (R)-1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)pyrrolidine-2-carboxylic acid dihydrochloride (12)

[0180]

[0181] Step 1: Synthesis of Compound 12-2

[0182] The solid-phase synthesis used in 12-2 was performed as follows: 1 g of 2-Cl-resin was weighed and placed in a peptide synthesis tube. 5 mL of dichloromethane was added, and the mixture was purged under nitrogen. Fmoc-D-proline (0.34 g, 1 mmol) and DIEA (0.26 g, 2 mmol) were added, and the mixture was purged under nitrogen for 1 h. 1 mL of methanol was added, and the mixture was purged under nitrogen for 30 min. The mixture was then dried under vacuum, washed three times with DMF (5 mL * 3), and dried under vacuum. This solution was used directly in the next step. 5 mL of piperidine / DMF (v:v = 1:4) was added to the peptide synthesis tube, and the mixture was purged under nitrogen for 30 min. The liquid was then dried under vacuum, washed three times with DMF (5 mL * 3), and dried under vacuum. This solution was used directly in the next step. 5 mL of piperidine / DMF (v:v = 1:4) was added to the peptide synthesis tube, and the mixture was purged under nitrogen for 30 min. The liquid was then dried under vacuum, washed three times with DMF (5 mL * 3), and dried under vacuum. This solution was used directly in the next step. DMF, nitrogen agitation, add Fmoc-D-Lys(Boc)-OH (0.94 g, 2 mmol), HATU (0.76 g, 2 mmol), DIEA (0.26 g, 2 mmol), nitrogen agitation for 1 h, drain the reaction solution, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, agitate with nitrogen for 30 min, drain the liquid, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, agitate with nitrogen for 30 min, drain the liquid, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube. DMF, nitrogen agitation, add Fmoc-D-Leu-OH (0.71 g, 2 mmol), HATU (0.76 g, 2 mmol), DIEA (0.26 g, 2 mmol), nitrogen agitation for 1 h, drain the reaction solution, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, agitate with nitrogen for 30 min, drain the liquid, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube. DMF, purged with nitrogen, added Fmoc-D-Phe-OH (0.77 g, 2 mmol), HATU (0.76 g, 2 mmol), DIEA (0.26 g, 2 mmol), purged with nitrogen for 1 h, dried under vacuum, washed 3 times with DMF (5 mL * 3), dried under vacuum, directly used for the next step; added 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, purged with nitrogen for 30 min, dried under vacuum, washed 3 times with DMF (5 mL * 3), dried under vacuum, directly used for the next step; added 50 mL DMF to the peptide synthesis tube, purged with nitrogen, added Boc-D-Phe-OH (0.53 g, 2 mmol), HATU (0.76 g, 2 mmol), DIEA (0.26 g, 2 mmol), DIEA (0.77 g, 2 mmol), DIEA (0.76 ...26 g (2 mmol), agitated under nitrogen for 1 h, the reaction solution was dried under vacuum, washed 3 times with DMF (5 mL * 3), dried under vacuum, washed 3 times with MeOH (5 mL * 3), dried under vacuum, washed 3 times with DCM (5 mL * 3), dried under vacuum and used directly for the next step; 5 mL of hexafluoroisopropanol / DCM (v:v = 1:4) was added to the polypeptide synthesis tube, agitated under nitrogen for 30 min, the liquid was dried under vacuum, and the process was repeated 3 times. The resulting liquids were combined and evaporated to dryness to obtain 12-2 as a white solid (0.57 g, yield: 66.4%), [M+H). + :851.5.

[0183] Step 2: Synthesis of Compound 12

[0184] Compound 12-2 (570 mg, 0.67 mmol) was dissolved in ethyl acetate (12 mL), and dioxane hydrochloride (4 M, 6 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 12 was a white solid (310 mg, yield: 64.0%), [M+H]. + :651.4.

[0185] 1 H NMR (400MHz, CD3OD), δ7.38-7.21(m,10H),4.73-4.64(m,2H),4.49-4.39(m,2H),4.15(m,1H),3.84(m,1H) ,3.48(m,1H),3.21(m,1H),3.05-2.97(m,4H),2.28(m,1H),2.08-1.52(m,13H),0.97(d,3H),0.94(d,3H).

[0186] Example 13

[0187] Synthesis of methyl(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)piperidin-4-yl)carbamate dihydrochloride (13)

[0188]

[0189] Step 1: Synthesis of Compound 13-2

[0190] 7-1 (1.0 g, 5.0 mmol) and triethylamine (1.0 g, 10 mmol) were dissolved in tetrahydrofuran (20 mL). 13-1 (0.94 g, 6.0 mmol) was then added to the solution under ice bath conditions. The reaction mixture was stirred overnight at room temperature. After the reaction was complete as monitored by TLC, the reaction mixture was filtered, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using eluent V. 正己烷 :V 乙酸乙酯 =From 15:1 to 5:1, a white solid 13-2 (1.39 g, yield: 86.9%), [M+H] was obtained. + :321.2.

[0191] Step 2: Synthesis of Compound 13-3

[0192] 13-2 (0.85 g, 2.66 mmol) and sodium methoxide (0.17 g, 3.19 mmol) were dissolved in methanol (10 mL). The reaction mixture was stirred overnight at 60 °C. After the reaction was completed by TLC monitoring, the reaction mixture was evaporated to dryness, dissolved in dichloromethane, and the organic phase was washed once with saturated ammonium chloride and once with saturated brine. The organic phase was dried and evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using eluent V. 二氯甲烷 :V 甲醇 The ratio of 1000:1 to 30:1 yielded a white solid 13-3 (0.57 g, yield: 83.2%), [M+H]. + :259.1. Step 3: Synthesis of compound 13-4

[0193] Compound 13-3 (0.57 g, 2.20 mmol) was dissolved in ethyl acetate (15 mL), and dioxane hydrochloride (4 M, 8 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 13-4 was a white solid (0.41 g, yield: 95.4%), [M+H]. + :159.1. Step 4: Synthesis of compound 13-6

[0194] 1-1 (350 mg, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) were dissolved in tetrahydrofuran (10 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 13-4 (99 mg, 0.51 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇The ratio of [M+H] was increased from 80:1 to 25:1 to obtain the target compound 13-6 as a white solid (198 mg, yield: 47.7%). + :894.4.

[0195] Step 5: Synthesis of Compound 13

[0196] Compound 13-6 (198 mg, 0.22 mmol) was dissolved in ethyl acetate (6 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 13 was a white solid (121 mg, yield: 71.1%), [M+H]. + :694.3.

[0197] 1 H NMR (400MHz, CD3OD), δ7.37-7.20(m,10H),4.87(m,1H),4.71(m,1H),4.40-4.28(m,2H),4.12(m,1H),4.02-3 .91(m,1H),3.63(m,4H),3.35-3.21(m,3H),3.02-2.80(m,5H),1.95-1.36(m,13H),0.98(d,3H),0.95(d,3H).

[0198] Example 14

[0199] Synthesis of N-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)piperidin-4-yl)-N-methyl-1H-indole-2-carboxamide dihydrochloride (14)

[0200]

[0201] Step 1: Synthesis of Compound 14-2

[0202] 6-1 (0.85 g, 3.97 mmol), HOBT (0.64 g, 4.76 mmol), EDCI (1.14 g, 5.96 mmol), and 14-1 (0.70 g, 4.37 mmol) were dissolved in DCM (25 mL). The reaction mixture was stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇The ratio of [M+H] was increased from 100:1 to 40:1 to obtain the target compound 14-2 as a white solid (1.10 g, yield: 77.6%). + :358.1.

[0203] Step 2: Synthesis of Compound 14-3

[0204] Compound 14-2 (1.10 g, 2.54 mmol) was dissolved in ethyl acetate (16 mL), and dioxane hydrochloride (4 M, 8 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 14-3 was a white solid (0.90 g, yield: 99.5%), [M+H]. + :258.1. Step 3: Synthesis of compound 14-5

[0205] 1-1 (350 mg, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) were dissolved in tetrahydrofuran (10 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 14-3 (150 mg, 0.51 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of alcohol to 100:1 was increased to 40:1 to give target compound 14-5 as a white solid (260 mg, yield: 56.4%), [M+H]. + :993.3.

[0206] Step 4: Synthesis of Compound 14

[0207] Compound 14-5 (260 mg, 0.26 mmol) was dissolved in ethyl acetate (6 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 14 was a white solid (125 mg, yield: 53.1%), [M+H]. + :793.3.

[0208] 1H NMR (400MHz, CD3OD), δ7.63-6.87(m,15H),4.69(m,2H),4.44(m,1H),4.18-4. 08(m,2H),3.34-2.75(m,13H),1.89-1.28(m,13H),0.98(d,3H),0.94(d,3H).

[0209] Example 15

[0210] Synthesis of N-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)piperidin-4-yl)-1H-indole-2-carboxamide dihydrochloride (15)

[0211]

[0212] Step 1: Synthesis of Compound 15-2

[0213] 7-1 (0.5 g, 2.50 mmol), HOBT (0.40 g, 3.0 mmol), EDCI (0.72 g, 3.75 mmol), and 14-1 (0.45 g, 2.80 mmol) were dissolved in DCM (25 mL). The reaction mixture was stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] was increased from 100:1 to 40:1 to obtain 15-2 (0.82 g, yield: 95.6%) of white solid. + :344.1.

[0214] Step 2: Synthesis of Compound 15-3

[0215] Compound 15-2 (0.82 g, 2.39 mmol) was dissolved in ethyl acetate (16 mL), and dioxane hydrochloride (4 M, 8 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 15-3 was a white solid (0.66 g, yield: 98.8%), [M+H]. + :244.1. Step 3: Synthesis of compound 15-5

[0216] 1-1 (250 mg, 0.33 mmol), HATU (152 mg, 0.40 mmol), and DIEA (150 mg, 1.16 mmol) were dissolved in tetrahydrofuran (10 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 15-3 (101 mg, 0.36 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] to [M+H] was increased from 100:1 to 40:1 to obtain the target compound 15-5 as a white solid (102 mg, yield: 31.4%). + :979.3.

[0217] Step 4: Synthesis of Compound 15

[0218] Compound 15-5 (102 mg, 0.10 mmol) was dissolved in ethyl acetate (5 mL), and dioxane hydrochloride (4 M, 2.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 15 was a white solid (77 mg, yield: 73.0%), [M+H]. + :779.3.

[0219] 1 H NMR (400MHz, CD3OD), δ7.60-6.89(m,15H),4.71(m,1H),4.55-4.40(m,2H),4.13(m,3H),3.39-3 .19(m,3H),3.04-2.83(m,5H),2.15-2.05(m,2H),1.84-1.42(m,13H),0.98(d,3H),0.94(d,3H).

[0220] Example 16

[0221] Synthesis of N-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)piperidin-4-yl)-N-methylimidazo[1,2-a]pyridine-2-carboxamide trihydrochloride (16)

[0222] Step 1: Synthesis of Compound 16-2

[0223] 6-1 (0.85 g, 3.97 mmol), HOBT (0.64 g, 4.76 mmol), EDCI (1.14 g, 5.96 mmol), and 16-1 (0.71 g, 4.37 mmol) were dissolved in DCM (25 mL). The reaction mixture was stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with saturated brine. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] was increased from 100:1 to 25:1 to obtain the target compound 16-2 as a white solid (0.91 g, yield: 64.0%). + :359.1.

[0224] Step 2: Synthesis of Compound 16-3

[0225] 16-2 (0.91 g, 2.54 mmol) was dissolved in ethyl acetate (16 mL), and dioxane hydrochloride (4 M, 8 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 16-3 was a white solid (0.72 g, yield: 85.8%), [M+H]. + :259.1.

[0226] Step 3: Synthesis of Compound 16-5

[0227] 1-1 (350 mg, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) were dissolved in tetrahydrofuran (10 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 16-3 (169 mg, 0.51 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with saturated brine. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] was increased from 100:1 to 25:1 to obtain the target compound 16-5 as a white solid (210 mg, yield: 45.4%). + :994.3.

[0228] Step 4: Synthesis of Compound 16

[0229] Compound 16-5 (210 mg, 0.21 mmol) was dissolved in ethyl acetate (6 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 16 was a white solid (113 mg, yield: 59.1%), [M+H]. + :794.3.

[0230] 1 H NMR (400MHz, CD3OD), δ8.87-8.77(m,2H),8.09(m,1H),7.96(m,1H),7.58(m,1H),7.36-7.21(m,10H),4.72(m,2H ),4.44(m,1H),4.14(m,2H),3.39-3.23(m,7H),3.05-2.82(m,6H),1.95-1.49(m,13H),0.99(d,3H),0.96(d,3H).

[0231] Example 17

[0232] Synthesis of (R)-N-((R)-6-amino-1-(3,5-dioxopiazin-1-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide dihydrochloride (17)

[0233]

[0234] Step 1: Synthesis of Compound 17-3

[0235] 1-1 (350 mg, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) were dissolved in tetrahydrofuran (10 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 17-2 (58 mg, 0.51 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] to [M+H] was increased from 100:1 to 40:1 to obtain the target compound 17-3 as a white solid (104 mg, yield: 26.4%). + :850.4.

[0236] Step 2: Synthesis of Compound 17

[0237] Compound 17-3 (104 mg, 0.12 mmol) was dissolved in ethyl acetate (4 mL), and dioxane hydrochloride (4 M, 2 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 17 was a white solid (75 mg, yield: 84.7%), [M+H]. + :650.3.

[0238] 1 H NMR(400MHz,CD3OD),δ7.37-7.22(m,10H),4.84(m,1H),4.75-4.69(m,1H),4.56-4.41(m,3H),4 .13(m,2H),3.37-3.21(m,3H),3.05-2.93(m,4H),1.81-1.42(m,9H),1.00(d,3H),0.97(d,3H).

[0239] Example 18

[0240] Synthesis of (R)-N-((R)-6-amino-1-(4-(2-(3,4-dichlorophenyl)acetamyl)piperidin-1-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide dihydrochloride (18)

[0241] Step 1: Synthesis of Compound 18-2

[0242] 7-1 (0.5 g, 2.5 mmol), HOBT (0.40 g, 3.0 mmol), EDCI (0.72 g, 3.75 mmol), and 2-2 (0.46 g, 2.27 mmol) were dissolved in DCM (15 mL). The reaction mixture was stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] to [M+H] was increased from 100:1 to 40:1 to obtain the target compound 18-2 as a white solid (0.93 g, yield: 96.1%). + :387.1.

[0243] Step 2: Synthesis of Compound 18-3

[0244] 18-2 (0.93 g, 2.40 mmol) was dissolved in ethyl acetate (16 mL), and dioxane hydrochloride (4 M, 8 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 18-3 was a white solid (0.77 g, yield: 99.0%), [M+H]. + :323.1. Step 3: Synthesis of compound 18-5

[0245] 1-1 (350 mg, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) were dissolved in tetrahydrofuran (10 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 18-3 (164 mg, 0.51 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] was increased from 100:1 to 40:1 to obtain the target compound 18-5 as a white solid (147 mg, yield: 31.0%). + :1022.4.

[0246] Step 4: Synthesis of Compound 18

[0247] Compound 18-5 (147 mg, 0.14 mmol) was dissolved in ethyl acetate (6 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 18 was a white solid (107 mg, yield: 83.0%), [M+H]. + :822.3.

[0248] 1 H NMR (400MHz, CD3OD), δ7.49-7.21(m,13H),4.86(m,1H),4.69(m,1H),4.46-4.31(m,2H),4.16-3.90(m,3H ),3.52-3.46(m,2H),3.25-3.18(m,2H),3.04-2.93(m,6H),1.95-1.28(m,13H),0.97(d,3H),0.93(d,3H).

[0249] Example 19

[0250] Synthesis of 4-amino-1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)-N-(4-chlorobenzyl)piperidine-4-carboxamide trihydrochloride (19)

[0251]

[0252] Step 1: Synthesis of Compound 19-2

[0253] 19-1 (350 mg, 0.36 mmol), HATU (163 mg, 0.43 mmol), and DIEA (93 mg, 0.72 mmol) were dissolved in tetrahydrofuran (8 mL). The reaction mixture was stirred at room temperature for 30 min. Then, p-chlorobenzylamine (57 mg, 0.40 mmol) was added, and the mixture was stirred overnight at room temperature. After the reaction was complete as monitored by TLC, the reaction mixture was washed once with 1N Na₂CO₃ and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] was increased from 100:1 to 40:1 to obtain the target compound 19-2 as a white solid (160 mg, yield: 40.6%). + :1103.4.

[0254] Step 2: Synthesis of Compound 19

[0255] Compound 19-2 (160 mg, 0.14 mmol) was dissolved in ethyl acetate (5 mL), and dioxane hydrochloride (4 M, 2 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 19 was a white solid (85 mg, yield: 64.2%), [M+H]. + :803.3.

[0256] 1 H NMR (400MHz, CD3OD), δ7.36-7.21(m,14H),4.84(m,1H),4.72-4.57(m,1H),4.43-4.34(m,3H),4.15(m,1H),3.61(m,1H ),3.35(m,1H),3.25-3.16(m,3H),3.04-2.95(m,4H),2.51-2.19(m,2H),1.98-1.48(m,12H),0.97(d,3H),0.93(d,3H).

[0257] Example 20

[0258] Synthesis of N-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)piperidin-4-yl)imidazo[1,2-a]pyridine-2-carboxamide trihydrochloride (20)

[0259]

[0260] Step 1: Synthesis of Compound 20-2

[0261] 7-1 (0.5 g, 2.5 mmol), HOBT (0.40 g, 3.0 mmol), EDCI (0.72 g, 3.75 mmol), and 16-1 (0.45 g, 2.80 mmol) were dissolved in DCM (25 mL). The reaction mixture was stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with saturated brine. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] was increased from 100:1 to 25:1 to obtain the target compound 20-2 as a brown solid (0.45 g, yield: 52.3%). + :345.1.

[0262] Step 2: Synthesis of compound 20-3

[0263] 20-2 (0.45 g, 1.31 mmol) was dissolved in ethyl acetate (12 mL), and dioxane hydrochloride (4 M, 10 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 20-3 was a white solid (0.35 g, yield: 95.4%), [M+H]. + :245.1. Step 3: Synthesis of compound 20-5

[0264] Dissolve 1-1 (250 mg, 0.33 mmol), HATU (152 mg, 0.40 mmol), and DIEA (150 mg, 1.16 mmol) in tetrahydrofuran (8 mL). Stir the reaction mixture at room temperature for 30 min. Then add 20-3 (101 mg, 0.36 mmol) and stir the mixture overnight at room temperature. After the reaction is complete as monitored by TLC, wash the reaction mixture once with 1N Na₂CO₃ and once with saturated brine. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using eluent V. 二氯甲烷 :V甲醇 The ratio was increased from 100:1 to 25:1, yielding a brown solid 20-5 (86 mg, yield: 26.4%), [M+H]. + :980.4.

[0265] Step 4: Synthesis of Compound 20

[0266] Compound 20-5 (86 mg, 0.09 mmol) was dissolved in ethyl acetate (5 mL), and dioxane hydrochloride (4 M, 2.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 20 was a white solid (69 mg, yield: 88.4%), [M+H]. + :780.4.

[0267] 1 H NMR (400MHz, CD3OD), δ8.92-8.79(m,2H),8.08(m,1H),7.94(m,1H),7.55(m,1H),7.36-7.16(m,10H),4.70(m,1H),4.63-4.39 (m,2H),4.23-4.11(m,2H),3.46-3.19(m,3H),3.07-2.88(m,5H),2.08(m,2H),1.87-1.42(m,13H),0.99(d,3H),0.95(d,3H).

[0268] Example 21

[0269] Synthesis of methyl (1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentaamino)hexylpiperidin-4-yl)(methyl)carbamate (21)

[0270]

[0271] Step 1: Synthesis of Compound 21-1

[0272] 6-1 (200 mg, 0.73 mmol) was dissolved in MeCN (3 mL), and K2CO3 (385 mg, 2.79 mmol) and methyl chloroformate (109 mg, 1.16 mmol) were added. The mixture was stirred overnight at room temperature. After the reaction was complete as monitored by TLC, the reaction solution was filtered, the organic phase was evaporated to dryness, and purified by column chromatography to obtain 21-1 as a colorless oil (220 mg, yield: 79.3%). [M+H] + :273.1.

[0273] Step 2: Synthesis of compound 21-2

[0274] 21-1 (220 mg, 0.80 mmol) was dissolved in DCM (3 mL), and dioxane hydrochloride (3 mL, 4 M) was added to the solution. The reaction was stirred for 1 hour. The reaction was stopped after the starting material was completely consumed by TLC, and the reaction solution was evaporated to dryness to obtain 21-2 as a white solid (150 mg, yield: 90.3%). [M+H] + :173.1.

[0275] Step 3: Synthesis of compound 21-3

[0276] Dissolve 1-1 (150 mg, 0.19 mmol) in THF (5 mL), then add HATU (91.3 mg, 0.24 mmol) and DIEA (77.4 mg, 0.60 mmol) to the solution. Stir the reaction at room temperature for 0.5 hours. Then add 21-2 (62 mg, 0.24 mmol), and continue stirring overnight. Stop the reaction when the starting material is consumed by TLC. Wash once with 1N Na₂CO₃ (10 mL) and once with 1N HCl (10 mL). Dry the organic phase and then evaporate to dryness to obtain the crude product. TLC of the crude product yields compound 21-3 as a brown solid (130 mg, yield: 71.8%). [M+H] + : 908.5.

[0277] Step 4: Synthesis of Compound 21

[0278] Compound 21-3 (130 mg, 0.14 mmol) was dissolved in DCM (1.5 mL), and TFA (0.5 mL) was added to the solution. The reaction was stirred at room temperature for 1 hour. When most of the starting material was consumed, the reaction was stopped by TLC. The reaction solution was directly concentrated, slurried with diethyl ether (10 mL), and the solid was filtered and dried to obtain the product. Target compound 21 was a grayish-white solid (53 mg, yield: 54.2%). [M+H] + :708.4.

[0279] Example 22

[0280] Synthesis of N-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)piperidin-4-yl)-N-methyl-3-oxo-[1,2,4]triazolo[4,3-a]pyridine-2(3H)-carboxamide dihydrochloride (22)

[0281]

[0282] Step 1: Synthesis of compound 22-2

[0283] Pyridinetriazolone 22-1 (1.50 g, 11.1 mmol) was weighed into a reaction flask, THF (20 mL) was added to the flask, and then triphosgene (3.63 g, 12.1 mmol) was added to the reaction solution. The mixture was then refluxed at 70 °C for 6 h. After cooling the reaction solution to room temperature, it was filtered, and the solid was dried to obtain a pale yellow solid 22-2 (2.12 g, yield: 68.0%).

[0284] Step 2: Synthesis of compound 22-3

[0285] 6-1 (1.50 g, 7.0 mmol) and triethylamine (1.41 g, 14.0 mmol) were dissolved in THF (20 mL). 22-2 (2.10 g, 7.5 mmol) was then added to the reaction solution. The reaction was stirred at 70 °C for 4 hours. After the reaction was complete as monitored by TLC, the solvent was evaporated and the solution was redissolved in DCM. The organic phase was washed once with 1N Na₂CO₃ and once with 1N HCl, and then dried by rotary evaporation to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] was increased from 100:1 to 40:1 to obtain a pale yellow solid 22-3 (2.45 g, yield: 93.2%). + :376.1.

[0286] Step 3: Synthesis of compound 22-4

[0287] Compound 22-3 (0.6 g, 1.6 mmol) was dissolved in ethyl acetate (12 mL), and dioxane hydrochloride (4 M, 8 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 22-4 was a pale yellow solid (0.49 g, yield: 98.3%), [M+H]. + :276.1.

[0288] Step 4: Synthesis of Compound 22-6

[0289] Dissolve 1-1 (250 mg, 0.33 mmol), HATU (152 mg, 0.40 mmol), and DIEA (150 mg, 1.16 mmol) in tetrahydrofuran (8 mL). Stir the reaction mixture at room temperature for 30 min. Then add 22-4 (112 mg, 0.36 mmol) and stir the mixture overnight at room temperature. After the reaction is complete as monitored by TLC, wash the reaction mixture once with 1N Na₂CO₃ and once with 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using eluent V.二氯甲烷 :V 甲醇 The ratio of [M+H] was increased from 100:1 to 35:1 to obtain the target compound 22-6 as a white solid (80 mg, yield: 23.9%). + :1011.3.

[0290] Step 5: Synthesis of Compound 22

[0291] Compound 22-6 (80 mg, 0.08 mmol) was dissolved in ethyl acetate (5 mL), and dioxane hydrochloride (4 M, 2.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 22 was a white solid (49 mg, yield: 70.0%), [M+H]. + :811.3.

[0292] 1 H NMR(400MHz,CD3OD),δ7.81(m,1H),7.33-7.16(m,12H),6.67(m,1H),4.73-4.66(m,2H),4.41(m,1H ),4.13(m,2H),3.34-3.15(m,4H),3.04-2.75(m,9H),1.94-1.38(m,13H),0.98(d,3H),0.94(d,3H).

[0293] Example 23

[0294] Synthesis of (2R)-N-((2R)-6-amino-1-(4-(3-benzyl-2-oxopyrrolidone-1-yl)piperidin-1-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropionamido)-3-phenylpropionamido)-4-methylpentanamide dihydrochloride (23)

[0295] Step 1: Synthesis of compound 23-2

[0296] 7-1 (1.50 g, 7.5 mmol) and triethylamine (1.52 g, 15.0 mmol) were dissolved in THF (30 mL). The reaction solution was placed in an ice bath. 4-Chlorobutyryl chloride (1.38 g, 9.8 mmol) was slowly added to the reaction solution. After the addition was complete, the reaction was stirred at room temperature for 2 hours. After the reaction was completed by TLC monitoring, the solvent was evaporated to dryness, redissolved in dichloromethane, and then washed once with 1N Na2CO3 and 1N HCl, respectively. The organic phase was dried, evaporated to dryness, and column chromatography was performed to obtain a pale yellow solid 23-2 (2.11 g, yield: 92.4%), [M+H]. +:305.3.

[0297] Step 2: Synthesis of compound 23-3

[0298] 23-2 (2.11 g, 6.93 mmol) was dissolved in THF (30 mL). The reaction solution was placed in an ice bath, and NaH (0.36 g, 9.01 mmol, 60% w / w) was slowly added to the reaction. The reaction solution was then refluxed overnight at 70 °C. When most of the starting material was consumed by TLC, the reaction was stopped. The reaction solution was poured into ice water, the organic solvent was removed by vortexing, and the mixture was extracted three times with DCM. The organic phases were combined, dried, and evaporated to dryness to obtain the crude product. The crude product was column chromatography to give a yellow solid 23-3 (1.70 g, yield: 91.5%), [M+H]. + :269.1. Step 3: Synthesis of compound 23-4

[0299] 23-3 (0.60 g, 2.23 mmol) was dissolved in dry THF (40 mL). The reaction solution was then purged with nitrogen and placed at -78 °C. LDA (2 M, 1.45 mL) was slowly added to the reaction solution, which was then stirred at -78 °C for 40 min. BnBr (0.50 g, 2.90 mmol) was then added to the reaction solution, and the mixture was slowly brought back to room temperature and stirred overnight. TLC monitoring showed that most of the starting material had been consumed, at which point the reaction was stopped. The reaction solution was poured into saturated ammonium chloride and extracted three times with DCM. The organic phases were combined, dried by rotary evaporation to obtain the crude product. The crude product was then column-purified to give a yellow solid 23-4 (0.37 g, yield: 46.2%), [M+H]. + :359.1. Step 4: Synthesis of compound 23-5

[0300] Compound 23-4 (0.37 g, 1.03 mmol) was dissolved in ethyl acetate (12 mL), and dioxane hydrochloride (4 M, 8 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 23-5 was a pale yellow solid (0.29 g, yield: 95.3%), [M+H]. + :259.1. Step 5: Synthesis of compound 23-7

[0301] Dissolve 1-1 (250 mg, 0.33 mmol), HATU (152 mg, 0.40 mmol), and DIEA (150 mg, 1.16 mmol) in tetrahydrofuran (8 mL). Stir the reaction mixture at room temperature for 30 min. Then add 23-5 (106 mg, 0.36 mmol) and stir the mixture overnight at room temperature. After the reaction is complete as monitored by TLC, wash the reaction mixture once with 1N Na₂CO₃ and once with saturated 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using eluent V. 二氯甲烷 :V 甲醇 The ratio of [M+H] was increased from 100:1 to 40:1 to obtain the target compound 23-7 as a white solid (86 mg, yield: 26.1%). + :994.3.

[0302] Step 6: Synthesis of Compound 23

[0303] 23-7 (86 mg, 0.09 mmol) was dissolved in ethyl acetate (5 mL), and dioxane hydrochloride (4 M, 2.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. When most of the starting material was consumed, the reaction was stopped by TLC. The reaction solution was directly filtered, and the solid was dried to give a white solid product 23 (60 mg, yield: 80.0%), [M+H]. + :794.3.

[0304] 1 H NMR(400MHz,CD3OD),δ7.33-7.20(m,15H),4.84(m,1H),4.69-4.59(m,2H),4.40(m,1H),4.12-3.97(m,2H) ,3.21-2.93(m,11H),2.91-2.69(m,4H),2.09-1.99(m,1H),1.95-1.45(m,15H),0.96(d,3H),0.93(d,3H).

[0305] Example 24

[0306] Synthesis of N-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)-4-methylpiperidin-4-yl)imidazo[1,2-a]pyridine-2-carboxamide trihydrochloride (24)

[0307] Step 1: Synthesis of Compound 24-2

[0308] 16-1 (0.29 g, 1.80 mmol), HATU (0.75 g, 1.97 mmol), and TEA (0.35 g, 3.44 mmol) were dissolved in DMF (6 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 24-1 (0.35 g, 1.64 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was diluted with water and extracted three times with DCM. The combined organic phases were washed once with 1N Na2CO3 and once with saturated brine, respectively. The organic phases were dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] to 100:1 was increased to 30:1 to obtain a brown solid 24-2 (0.53 g, yield: 90.5%). + :359.1.

[0309] Step 2: Synthesis of compound 24-3

[0310] Compound 24-2 (0.53 g, 1.48 mmol) was dissolved in ethyl acetate (12 mL), and dioxane hydrochloride (4 M, 8 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 24-3 was a white solid (0.48 g, yield: 98.0%), [M+H]. + :259.1.

[0311] Step 3: Synthesis of Compound 24-5

[0312] Dissolve 1-1 (250 mg, 0.33 mmol), HATU (152 mg, 0.40 mmol), and DIEA (150 mg, 1.16 mmol) in tetrahydrofuran (8 mL). Stir the reaction mixture at room temperature for 30 min. Then add 24-3 (119 mg, 0.36 mmol) and stir the mixture overnight at room temperature. After the reaction is complete as monitored by TLC, wash the reaction mixture once with 1N Na₂CO₃ and once with saturated brine. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using eluent V. 二氯甲烷 :V 甲醇 =From 100:1 to 25:1, a white solid 24-5 (110 mg, yield: 33.3%) was obtained, [M+H]. + :994.3.

[0313] Step 4: Synthesis of Compound 24

[0314] Compound 24-5 (110 mg, 0.11 mmol) was dissolved in ethyl acetate (5 mL), and dioxane hydrochloride (4 M, 2.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 24 was a white solid (89 mg, yield: 89.0%), [M+H]. + :794.3.

[0315] 1 H NMR (400MHz, CD3OD), δ8.93-8.82(m,2H),8.04(m,1H),7.92(m,1H),7.53(m,1H),7.34-7.13(m,10H),4.73-4.63(m,1H),4.44-4.36(m,1H ),4.14(m,1H),4.00-3.56(m,2H),3.40-3.14(m,3H),3.04-2.93(m,4H),2.54-2.35(m,2H),1.79-1.28(m,16H),0.99(d,3H),0.96(d,3H).

[0316] Example 25

[0317] Synthesis of N-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)-4-methylpiperidin-4-yl)-N-methylimidazo[1,2-a]pyridine-2-carboxamide trihydrochloride (25)

[0318] Step 1: Synthesis of Compound 25-3

[0319] 24-2 (0.32 g, 0.89 mmol) was dissolved in DMF (4 mL). The reaction solution was placed in an ice bath. NaH (46 mg, 1.16 mmol, 60% w / w) was added to the reaction solution, and the solution was stirred at room temperature for 30 min. Then, CH3I (202 mg, 1.42 mmol) was added, and the solution was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction solution was poured into ice water, and the aqueous phase was extracted three times with DCM. The organic phases were combined and dried by rotary evaporation to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 =100:1 to 30:1 yielded a light brown solid 25-3 (0.28 g, yield: 84.2%), [M+H] + :373.1.

[0320] Step 2: Synthesis of compound 25-4

[0321] 25-3 (0.28 g, 0.75 mmol) was dissolved in tetrahydrofuran (8 mL), and dioxane hydrochloride (4 M, 8 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 25-4 was a white solid (0.24 g, yield: 92.4%), [M+H]. + :273.1.

[0322] Step 3: Synthesis of Compound 25-6

[0323] Dissolve 1-1 (250 mg, 0.33 mmol), HATU (152 mg, 0.40 mmol), and DIEA (150 mg, 1.16 mmol) in tetrahydrofuran (8 mL). Stir the reaction mixture at room temperature for 30 min. Then add 25-4 (124 mg, 0.36 mmol) and stir overnight at room temperature. After the reaction is complete as monitored by TLC, wash the reaction mixture once with 1N Na₂CO₃ and once with saturated brine. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using eluent V. 二氯甲烷 :V 甲醇 =100:1 to 25:1 to give a white solid 25-6 (125 mg, yield: 37.4%), [M+H] + :1008.4.

[0324] Step 4: Synthesis of Compound 25

[0325] Compound 25-6 (125 mg, 0.12 mmol) was dissolved in ethyl acetate (5 mL), and dioxane hydrochloride (4 M, 2.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered and the solid was dried to obtain the product. Target compound 25 was a white solid (88 mg, yield: 77.3%), [M+H]. + :808.3.

[0326] 1H NMR (400MHz, CD3OD), δ8.83-8.67(m,2H),8.05(m,1H),7.94(m,1H),7.53(m,1H),7.34-7.16(m,10H),4.66(m,1H),4.38(m,1H),4.1 4-4.04(m,2H),3.82(m,1H),3.34-3.21(m,6H),3.04-2.94(m,4H),2.71-2.43(m,2H),1.85-1.29(m,15H),0.99(d,3H),0.92(d,3H).

[0327] Example 26

[0328] Synthesis of methyl(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)-4-methylpiperidin-4-yl)carbamate dihydrochloride (26)

[0329]

[0330] Step 1: Synthesis of Compound 26-2

[0331] 24-1 (0.35 g, 1.6 mmol) and triethylamine (0.32 g, 3.2 mmol) were dissolved in tetrahydrofuran (16 mL). 13-1 (0.31 g, 2.0 mmol) was then added to the solution under ice bath conditions. The reaction mixture was stirred overnight at room temperature. After the reaction was complete as monitored by TLC, the reaction mixture was filtered, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using eluent V. 正己烷 :V 乙酸乙酯 The ratio of [M+H] to [5] was adjusted from 15:1 to 5:1 to obtain the target compound 26-2 as a white solid (0.50 g, yield: 91.5%). + :335.1.

[0332] Step 2: Synthesis of compound 26-3

[0333] 26-2 (0.50 g, 1.5 mmol) and sodium methoxide (92 mg, 1.7 mmol) were dissolved in methanol (10 mL). The reaction mixture was then stirred overnight at 60 °C. After the reaction was completed by TLC monitoring, the reaction mixture was evaporated to dryness, dissolved in dichloromethane, and the organic phase was washed once with saturated ammonium chloride and once with saturated brine. The organic phase was dried and evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using eluent V. 二氯甲烷 :V 甲醇The ratio of [M+H] to [M+H] was increased from 100:1 to 40:1 to obtain the target compound 26-3 as a white solid (0.39 g, yield: 95.8%). + :273.1.

[0334] Step 3: Synthesis of compound 26-4

[0335] Compound 26-3 (0.39 g, 1.4 mmol) was dissolved in ethyl acetate (12 mL), and dioxane hydrochloride (4 M, 8 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 26-4 was a white solid (0.29 g, yield: 97.0%), [M+H]. + :173.1.

[0336] Step 4: Synthesis of compound 26-6

[0337] 1-1 (250 mg, 0.33 mmol), HATU (152 mg, 0.40 mmol), and DIEA (150 mg, 1.16 mmol) were dissolved in tetrahydrofuran (8 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 26-4 (75 mg, 0.36 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 =80:1 to 25:1 to give white solid 26-6 (150 mg, yield: 49.8%), [M+H] + :908.3.

[0338] Step 5: Synthesis of Compound 26

[0339] Compound 26-6 (150 mg, 0.17 mmol) was dissolved in ethyl acetate (6 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 26 was a white solid (102 mg, yield: 79.1%), [M+H]. + :708.2.

[0340] 1H NMR(400MHz,CD3OD),δ7.35-7.19(m,10H),4.84(m,1H),4.71(m,1H),4.39(m,1H),4.14-3.93(m,2H),3.74(m,1H),3 .58(s,3H),3.40-3.07(m,3H),3.01-2.91(m,4H),2.24-2.03(m,2H),1.78-1.29(m,15H),0.98(d,3H),0.94(d,3H).

[0341] Example 27

[0342] Synthesis of (R)-N-((R)-6-amino-1-oxo-1-(3-oxo-2,8-diazaspiro[4.5]decane-8-yl)hex-2-yl)-4-methyl-2-((R)-3-phenyl-2-(2-((R)-2-phenylpropyl)amino)acetamyl)propamido)pentanamide ditrifluoroacetate (27)

[0343] Step 1: Synthesis of Compound 27-3

[0344] 27-1 (148 mg, 0.19 mmol), HATU (87 mg, 0.23 mmol), and DIEA (62 mg, 0.48 mmol) were dissolved in tetrahydrofuran (6 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 8-2 (38 mg, 0.20 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, DCM was added to the reaction mixture, and the solution was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] was increased from 100:1 to 30:1 to obtain the target compound 27-3 as a white solid (122 mg, yield: 70.1%). + :918.3.

[0345] The solid-phase synthesis used in 27-1 was performed as follows: 1 g of 2-Cl-resin was weighed and placed in a peptide synthesis tube. 5 mL of dichloromethane was added, and the mixture was purged under nitrogen. Fmoc-D-Lys(Boc)-OH (0.47 g, 1 mmol) and DIEA (0.26 g, 2 mmol) were added, and the mixture was purged under nitrogen for 1 h. 1 mL of methanol was added, and the mixture was purged under nitrogen for 30 min. The mixture was then dried under vacuum, washed three times with DMF (5 mL * 3), and dried under vacuum. This mixture was used directly in the next step. 5 mL of piperidine / DMF (v:v = 1:4) was added to the peptide synthesis tube, and the mixture was purged under nitrogen for 30 min. The liquid was then dried under vacuum, washed three times with DMF (5 mL * 3), and dried under vacuum. This mixture was used directly in the next step. 5 mL of piperidine / DMF (v:v = 1:4) was added to the peptide synthesis tube, and the mixture was purged under nitrogen for 30 min. The liquid was then dried under vacuum, washed three times with DMF (5 mL * 3), and dried under vacuum. This mixture was used directly in the next step. DMF, nitrogen agitation, add Fmoc-D-Leu-OH (0.71 g, 2 mmol), HATU (0.76 g, 2 mmol), DIEA (0.26 g, 2 mmol), nitrogen agitation for 1 h, drain the reaction solution, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, agitate with nitrogen for 30 min, drain the liquid, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube. DMF, nitrogen agitation, add Fmoc-D-Phe-OH (0.77 g, 2 mmol), HATU (0.76 g, 2 mmol), DIEA (0.26 g, 2 mmol), nitrogen agitation for 1 h, drain the reaction solution, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, agitate with nitrogen for 30 min, drain the liquid, wash 3 times with DMF (5 mL * 3), drain directly for the next step; add 5 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube. DMF was added under nitrogen agitation, followed by (R)-2-((tert-Butoxycarbonyl)(2-phenylpropyl)amino)acetic acid (0.58 g, 2 mmol), HATU (0.76 g, 2 mmol), and DIEA (0.26 g, 2 mmol). The mixture was agitated under nitrogen for 1 h, and the reaction solution was dried. The solution was washed three times with DMF (5 mL x 3), dried, and then washed three times with MeOH (5 mL x 3). The solution was dried and then washed three times with DCM (5 mL x 3). The solution was then dried and used directly in the next step. 5 mL of hexafluoroisopropanol / DCM (v:v = 1:4) was added to the polypeptide synthesis tube, and the mixture was agitated under nitrogen for 30 min. The solution was dried, and this process was repeated three times. The resulting liquids were combined and evaporated to dryness to obtain 1-1 as a white solid (0.61 g, yield: 78.0%), [M+H]. + :782.4.

[0346] Step 2: Synthesis of Compound 27

[0347] Compound 27-3 (122 mg, 0.13 mmol) was dissolved in dichloromethane (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was evaporated to dryness, slurried with diethyl ether, and the solid was dried to obtain the product. Target compound 27 was a white solid (112 mg, yield: 89.1%), [M+H]. + :718.2.

[0348] 1 H NMR (400MHz, CD3OD), δ7.36-7.13(m,10H),4.70(m,1H),4.58(m,1H),4.35(m,1H),3.91-3.44 (m,6H),3.21-2.84(m,9H),2.31-2.24(m,2H),1.66-1.30(m,16H),0.93(d,3H),0.89(d,3H).

[0349] Example 28

[0350] Synthesis of (R)-N-((R)-1-(2-acetyl-2,8-diazaspiro[4.5]decane-8-yl)-6-amino-1-oxohex-2-yl)-4-methyl-2-((R)-3-phenyl-2-(2-((R)-2-phenylpropyl)amino)acetamyl)propylamino)pentanoic acid ditrifluoroacetate (28)

[0351] Step 1: Synthesis of compound 28-2

[0352] 28-1 (0.30 g, 1.25 mmol) and triethylamine (0.25 g, 2.50 mmol) were dissolved in THF (10 mL). The reaction mixture was placed in an ice bath. Acetyl chloride (0.14 g, 1.75 mmol) was slowly added dropwise to the reaction mixture. The reaction was stirred at room temperature for 1 hour. After the reaction was completed by TLC monitoring, the reaction mixture was evaporated to dryness, redissolved in ethyl acetate, and then washed once each with 1N Na₂CO₃ and 1N HCl. The organic phase was dried and evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] to [M+H] was increased from 100:1 to 30:1 to obtain the target compound 28-2 as a yellow oil (0.31 g, yield: 87.9%). + :283.1.

[0353] Step 2: Synthesis of compound 28-3

[0354] Compound 28-2 (0.31 g, 1.10 mmol) was dissolved in dichloromethane (9 mL), and trifluoroacetic acid (3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 3 hours. When most of the starting material was consumed, the reaction was stopped, and the reaction mixture was directly evaporated to dryness and then slurried with diethyl ether to obtain the product. The target compound 28-3 was a white solid (0.29 g, yield: 89.1%), [M+H]. + :183.1. Step 3: Synthesis of compound 28-5

[0355] 27-1 (150 mg, 0.19 mmol), HATU (87 mg, 0.23 mmol), and DIEA (73 mg, 0.57 mmol) were dissolved in tetrahydrofuran (8 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 28-3 (62 mg, 0.21 mmol) was added, and the mixture was stirred overnight at room temperature. After the reaction was complete as monitored by TLC, the reaction mixture was washed once with 1N Na₂CO₃ and once with 1N HCl. The organic phase was dried by rotary evaporation to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] to [M+H] was increased from 100:1 to 30:1 to obtain the target compound 28-5 as a white solid (124 mg, yield: 68.3%). + :946.3.

[0356] Step 4: Synthesis of Compound 28

[0357] 28-5 (124 mg, 0.13 mmol) was dissolved in dichloromethane (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. When most of the starting material was consumed, the reaction was stopped, and the reaction solution was directly evaporated to dryness and then slurried with diethyl ether to obtain the product, a white solid 28 (103 mg, yield: 80.7%), [M+H]. + :746.3.

[0358] 1 H NMR(400MHz,CD3OD),δ7.38-7.13(m,10H),4.70(m,1H),4.57(m,1H),4.37(m,1H),3.94-3.35 (m,10H),3.23-3.04(m,4H),2.94-2.80(m,3H),2.06-1.30(m,21H),0.95(d,3H),0.91(d,3H).

[0359] Example 29

[0360] Synthesis of (R)-N-((R)-6-amino-1-(2,4-dioxo-1,3,8-triazaspiro[4.5]decane-8-yl)-1-oxohex-2-yl)-4-methyl-2-((R)-3-phenyl-2-(2-((R)-2-phenylpropyl)amino)acetamyl)propionamide)pentanamide ditrifluoroacetate (29)

[0361] Step 1: Synthesis of Compound 29-3

[0362] 27-1 (148 mg, 0.19 mmol), HATU (87 mg, 0.23 mmol), and DIEA (62 mg, 0.48 mmol) were dissolved in tetrahydrofuran (6 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 29-2 (41 mg, 0.20 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, DCM was added to the reaction mixture, and the solution was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 =100:1 to 30:1 to give white solid 29-3 (116 mg, yield: 70.1%), [M+H] + :933.5.

[0363] Step 2: Synthesis of Compound 29

[0364] Compound 29-3 (116 mg, 0.12 mmol) was dissolved in dichloromethane (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. When most of the starting material was consumed, the reaction was stopped by TLC. The reaction mixture was evaporated to dryness, slurried with diethyl ether, and the solid was dried to obtain the product. Target compound 29 was a white solid (105 mg, yield: 87.9%), [M+H]. + :733.4.

[0365] Example 30

[0366] Synthesis of (R)-6-amino-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamino)-N-(1-(2-(3,4-dichlorophenyl)acetyl)piperidin-4-yl)-N-methylhexylamine hydrochloride (30)

[0367] Step 1: Synthesis of Compound 30-2

[0368] 2-2 (2 g, 9.76 mmol), 30-1 (1.79 g, 11.71 mmol), HOBT (1.58 g, 11.71 mmol), and EDCI·HCl (2.8 g, 14.64 mmol) were dissolved in 5 mL of dichloromethane. After stirring at room temperature for 5 min, triethylamine (0.99 g, 9.76 mmol) was added, and the mixture was stirred at room temperature overnight. The reaction was quenched with saturated sodium bicarbonate aqueous solution, and the organic phase was separated. The aqueous phase was extracted three times with dichloromethane. The combined organic phases were washed with 1N dilute hydrochloric acid, concentrated, and subjected to column chromatography (V). PE :V EA =1:1), yielding 30-2 (1.2g, yield 42%) of oily substance.

[0369] Step 2: Synthesis of compound 30-3

[0370] Dissolve 30-2 (1.2 g, 4.21 mmol) in 20 mL of THF, stir at room temperature for 10 min, add methylamine-methanol solution (0.65 g, 6.31 mmol), stir for 0.5 hours, then add sodium triacetoxyborohydride (2.22 g, 10.53 mmol), stir for 4 hours. Quench the reaction with saturated sodium carbonate solution, separate the organic phase, and use V... DCM :V MeOH Extracted three times at a ratio of 10:1, the organic phases were combined and concentrated to obtain crude product 30-3 (1.5g).

[0371] Step 3: Synthesis of compound 30-4

[0372] Dissolve 1-1 (500 mg, 0.664 mmol) and HATU (317 mg, 0.797 mmol) in THF (10 mL), then add DIEA (269 mg, 1.992 mmol). Stir the reaction at room temperature for 0.5 hours, then add 30-3 (250 mg, 0.797 mmol) and stir the reaction at room temperature for 7 hours. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and use V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =50:1), yielded a white solid 30-4 (400 mg, yield 46%).

[0373] Step 4: Synthesis of Compound 30

[0374] Compound 30-4 (400 mg, 0.386 mmol) was dissolved in 10 mL of ethyl acetate, and then 5 mL of dioxane hydrochloride solution was added. The mixture was stirred at room temperature for 1.5 hours, and then filtered to obtain the product. Target compound 30 was a white solid (150 mg, yield: 83%), [M+H]. + :836.8.

[0375] 1 H NMR(400MHz,CD3OD),δ7.46-7.22(m,13H),4.73-4.39(m,3H),4.11-4.05(m,2H),2.93-2.7 7(m,5H),2.03(d,1H),1.68-1.61(m,10H),1.38(dd,2H),1.31-1.29(m,3H),0.96(dd,6H).

[0376] Example 31

[0377] Synthesis of (R)-1-((5R,8R,11R,14R)-14-amino-5-(4-aminobutyl)-11-benzyl-8-isobutyl-3-methyl-4,7,10,13-tetraoxy-15-phenyl-3,6,9,12-tetraazapentadecanyl)pyrrolidine-2-carboxylic acid hydrochloride (31)

[0378] Step 1: Synthesis of compound 31-2

[0379] Dissolve 31-1a (500 mg, 3.05 mmol) in dichloromethane, add 31-1b (785 mg, 4.55 mmol), stir at room temperature for 0.5 hours, add NaB(AcO)3 (1.605 g, 7.6 mmol), and continue stirring for 3 hours. Quench the reaction with water, separate the organic phase, extract the aqueous phase three times with dichloromethane, combine the organic phases, concentrate, and perform column chromatography (V). PE :V EA =1:1), yielding oily substance 31-2 (400 mg, yield 57.8%).

[0380] Step 2: Synthesis of compound 31-3

[0381] Dissolve 31-2 (400 mg, 1.40 mmol) in 5 mL of ethyl acetate, then add 2.5 mL of dioxane hydrochloride solution and stir at room temperature for 1.5 hours. Discard the supernatant to obtain a white paste, 31-3 (300 mg).

[0382] Step 3: Synthesis of compound 31-4

[0383] Dissolve 1-1 (250 mg, 0.332 mmol) and HATU (151.4 mg, 0.398 mmol) in 5 mL of THF, then add DIEA (171.3 mg, 1.328 mmol). Stir the reaction at room temperature for 0.5 hours, then add 31-3 (88.4 mg, 0.398 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 31-4 (100mg).

[0384] Step 4: Synthesis of Compounds 31-5

[0385] Dissolve 31-4 (130 g, 0.141 mmol) in 2 mL of THF, add NaOH aqueous solution (1 mmol / mL), stir at room temperature for 5 hours, separate the organic phase, and obtain crude product 31-5.

[0386] Step 5: Synthesis of Compound 31

[0387] The crude product 31-5 was dissolved in 2 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 1.5 hours, and then filtered to obtain the product. Target compound 31 was a white solid (100 mg, yield: 83%), [M+H]. + :708.96.

[0388] 1 H NMR (400MHz, CD3OD) δ7.38-7.29(m,10H),4.73-4.71(m,1),4.44-4.42(m,2H),4.18-4.16(m,1H),3.88-3.86(m,1H),3.33 -3.31(m,3H),3.03-3.01(m,4H),2.58-2.56(m,1H),2.2-2.20(m,1H),1.73-1.63(m,9H),1.33-1.31(m,2H),0.99(dd,6H).

[0389] Example 32

[0390] Synthesis of (R)-N-((R)-6-amino-1-(4-(N-methylcinnamamido)piperidin-1-yl)-1-oxohexane-2-yl)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide hydrochloride (32)

[0391] Step 1: Synthesis of compound 32-2

[0392] Dissolve 32-1 (259 mg, 1.75 mmol), 6-1 (450 mg, 2.1 mmol), HOBT (285 mg, 2.1 mmol), and EDCI.HCl (503 mg, 2.625 mmol) in 10 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (176 mg, 1.75 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, concentrate, and perform column chromatography (V... PE :V EA =5:1), to obtain oily substance 32-2 (440mg, yield 42%).

[0393] Step 2: Synthesis of compound 32-3

[0394] Dissolve 32-2 (440 mg, 1.28 mmol) in 10 mL of ethyl acetate, then add dioxane hydrochloride (4 M, 8 mL), stir at room temperature for 1 hour, and filter to obtain a white solid 32-3 (240 mg, 67%).

[0395] Step 3: Synthesis of compound 32-4

[0396] Dissolve 1-1 (250 mg, 0.332 mmol) and HATU (151.4 mg, 0.398 mmol) in 5 mL of THF, then add DIEA (171.3 mg, 1.328 mmol). Stir the reaction at room temperature for 0.5 hours, then add 32-3 (111.5 mg, 0.398 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and use V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 32-4 (270 mg, yield 83%).

[0397] Step 4: Synthesis of Compound 32

[0398] Dissolve 32-4 (270 mg, 0.276 mmol) in 5 mL of ethyl acetate, add 2.5 mL of dioxane hydrochloride solution, stir at room temperature for 1.5 hours, filter, and obtain the product product, a brown solid 32-4 (100 mg, yield 42%), [M+H]. + :781.

[0399] 1 H NMR(400MHz,CD3OD)δ7.59-7.09(m,15H),4.71(m,3),4.43(m,2H),4.14(m,2H) ,3.24(m,2H),3.01(m,6H),2.9(s,6H),1.67(m,14H),1.33(m,2H),0.99(dd,6H)

[0400] Example 33

[0401] Synthesis of (R)-N-((R)-6-amino-1-oxo-1-(3-oxoperazin-1-yl)hexane-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide hydrochloride (33)

[0402]

[0403] Step 1: Synthesis of compound 33-2

[0404] Dissolve 1-1 (250 mg, 0.332 mmol) and HATU (151.2 mg, 0.398 mmol) in THF. Add DIEA and stir at room temperature for half an hour. Then add piperazine 33-1 (39.8 mg, 0.398 mmol) and stir at room temperature overnight. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and use V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 33-2 (240mg).

[0405] Step 2: Synthesis of Compound 33

[0406] Compound 33-2 (240 mg, 0.287 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride solution was added. The mixture was stirred at room temperature for 1.5 hours, and then filtered to obtain the product. Target compound 33 was a white solid (80 mg), [M+H]. + :636.4.

[0407] 1H NMR (400MHz, CD3OD) δ7.38-7.29(m,10H),4.73-4.71(m,1H),4.42-4.39(m,1H),4.18-4.16(m, 1H),3.88-3.86(m,1H),3.29-3.21(m,1H),3.03-3.01(m,4H),1.73-1.63(m,9H),0.99(dd,6H).

[0408] Example 34

[0409] Synthesis of N-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanyl)hexanoyl)piperidin-4-yl)-N-methylbenzofuran-5-carboxamide hydrochloride (34)

[0410] Step 1: Synthesis of compound 34-2

[0411] Dissolve 34-1 (189.3 mg, 1.168 mmol), 6-1 (300 mg, 1.40 mmol), HOBT (189 mg, 1.40 mmol), and EDCI.HCl (335.9 mg, 1.75 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (118.2 mg, 1.168 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, concentrate, and perform column chromatography (V). PE :V EA =5:1), yielding an oily substance 34-2 (360 mg, yield 72%).

[0412] Step 2: Synthesis of compound 34-3

[0413] Dissolve 34-2 (360 mg, 1.006 mmol) in 5 mL of ethyl acetate, add 2.5 mL of dioxane hydrochloride solution, stir at room temperature for 4 hours, filter, and obtain white solid 34-3.

[0414] Step 3: Synthesis of compound 34-4

[0415] Dissolve 1-1 (200 mg, 0.266 mmol) and HATU (121.22 mg, 0.319 mmol) in THF (5 mL), then add DIEA (137 mg, 1.06 mmol). Stir the reaction at room temperature for 0.5 hours, then add 34-3 (93.79 mg, 0.319 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 34-4 (180 mg, yield 66%).

[0416] Step 4: Synthesis of Compound 34

[0417] Compound 34-4 (180 mg) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride solution was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. Target compound 34 was a white solid (100 mg, yield 66.2%), [M+H]. + :794.5.

[0418] 1 H NMR (400MHz, CD3OD) δ7.76-7.74(m,1H),7.52-7.50(m,1H),7.38-7.29(m,13H),4.73-4.71(m,2 H),4.42-4.39(m,1H),3.29-3.21(m,3H),3.03-3.01(m,5H),1.73-1.63(m,13H),0.99(dd,6H).

[0419] Example 35

[0420] Synthesis of N-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanyl)hexanoyl)piperidin-4-yl)-N-methylbenzofuran-2-carboxamide hydrochloride (35)

[0421] Step 1: Synthesis of Compound 35-2

[0422] Dissolve 35-1 (189.3 mg, 1.168 mmol), 6-1 (300 mg, 1.40 mmol), HOBT (189 mg, 1.40 mmol), and EDCI.HCl (335.9 mg, 1.75 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (118.2 mg, 1.168 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, concentrate, and perform column chromatography (V). PE :V EA =5:1), yielding an oily substance 35-2 (360mg, yield 72%).

[0423] Step 2: Synthesis of compound 35-3

[0424] Dissolve 35-2 (360 mg, 1.006 mmol) in 5 mL of ethyl acetate, add 2.5 mL of dioxane hydrochloride, stir at room temperature for 4 hours, filter, and obtain white solid 35-3.

[0425] Step 3: Synthesis of compound 35-4

[0426] Dissolve 1-1 (200 mg, 0.266 mmol) and HATU (121.22 mg, 0.319 mmol) in THF (5 mL), then add DIEA (137 mg, 1.06 mmol). Stir the reaction at room temperature for 0.5 hours, then add 35-3 (93.79 mg, 0.319 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 35-4 (190 mg, yield 70%).

[0427] Step 4: Synthesis of Compound 35

[0428] Compound 35-4 (180 mg, 0.181 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. Target compound 35 was a white solid (100 mg, yield 65.4%), [M+H]. + :794.4.

[0429] 1H NMRδ7.89(s,1H),7.74-7.61(m,2H),7.38-7.29(m,11H),6.9(s,1H),4.73-4.71 (m,2H),4.42-4.39(m,1H),3.03-3.01(m,8H),1.73-1.63(m,13H),0.99(dd,6H).

[0430] Example 36

[0431] Synthesis of (R)-1-((5R,8R,11R,14R)-14-amino-5-(4-aminobutyl)-11-benzyl-8-isobutyl-3-methyl-4,7,10,13-tetraoxy-15-phenyl-3,6,9,12-tetraazapentadecanyl)-N-methylpyrrolidine-2-carboxamide hydrochloride (36)

[0432] Step 1: Synthesis of Compound 36-2

[0433] Dissolve 31-4 (83 mg, 0.09 mmol) in 1 mL of THF, then add 1 mL of methylamine aqueous solution (30%). Stir the mixture at room temperature for 24 hours. TLC monitoring showed incomplete reaction, so the temperature was increased to 40 °C and the reaction continued for another 24 hours. Concentrate and then perform column chromatography (V... DCM :V MeOH =20:1), yielding product 36-2.

[0434] Step 2: Synthesis of Compound 36

[0435] The product 36-2 was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride solution was added. The mixture was stirred at room temperature for 2 hours, filtered, and the product was obtained. The target compound 36 was a white solid (56 mg, two-step yield 86.1%), [M+H]. + :721.02.

[0436] 1 H NMR(400MHz,CD3OD)δ7.30-7.20(m,10H),4.75-4.71(m,2H),4.45-4.43(m,1H),4.27- 4.13(m,2H),3.21-3.18(m,2H),2.99-2.17(m,9H),1.73-1.63(m,13H),0.99(dd,6H).

[0437] Example 37

[0438] Synthesis of 2-((1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanyl)hexanoyl)piperidin-4-yl)amino)-2-oxoacetic acid hydrochloride (37)

[0439]

[0440] Step 1: Synthesis of Compound 37-2

[0441] Dissolve 37-1 (220.3 mg, 1.18 mmol) and 7-1 (300 mg, 1.50 mmol) in 5 mL of dichloromethane, add triethylamine (181.98 mg, 1.8 mmol), and stir at room temperature for 1.5 hours. Quench the reaction with water, extract three times with dichloromethane, and concentrate to give a white solid 37-2 (510 mg, yield 99.2%).

[0442] Step 2: Synthesis of compound 37-3

[0443] Dissolve 37-2 (350 mg, 0.875 mmol) in 5 mL of ethyl acetate, add 3 mL of dioxane hydrochloride, stir at room temperature for 4 hours, filter, and obtain white solid 37-3.

[0444] Step 3: Synthesis of compound 37-4

[0445] Dissolve 1-1 (150 mg, 0.199 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 37-3 (53.2 mg, 0.239 mmol) and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 37-4 (130 mg, yield 69.5%).

[0446] Step 4: Synthesis of compound 37-5

[0447] 37-4 (130 mg, 0.141 mmol) was dissolved in 1 mL of THF, and then 1 mL of NaOH aqueous solution (1 N) was added. The mixture was stirred at room temperature for 3 hours. The THF was concentrated to dryness, and the aqueous phase was separated into two phases. DCM :V MeOHExtracted three times at a ratio of 10:1, concentrated to obtain product 37-5 (80 mg, yield 62.5%).

[0448] Step 5: Synthesis of Compound 37

[0449] Compound 37-5 (80 mg, 0.088 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride solution was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. The target compound 37 was a white solid (50 mg, yield 72.8%), [M+H]. + :708.94.

[0450] 1 H NMR(400MHz,CD3OD)δ7.37-7.31(m,10H),4.73-4.71(m,1H),4.42-4.39(m,2H),4.16 -4.13(m,2H),3.33-3.31(m,2H),3.03-3.01(m,5H),1.73-1.63(m,13H),0.99(dd,6H)

[0451] Example 38

[0452] Synthesis of (R)-N-((R)-6-amino-1-(4-(N-methyl-2-(4-(trifluoromethyl)phenyl)acetamide)piperidin-1-yl)-1-oxohexane-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide (38)

[0453]

[0454] Step 1: Synthesis of Compound 38-2

[0455] Dissolve 38-1 (238.27 mg, 1.168 mmol), 6-1 (300 mg, 1.40 mmol), HOBT (189 mg, 1.40 mmol), and EDCI.HCl (335.9 mg, 1.75 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (118.2 mg, 1.168 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, concentrate, and perform column chromatography (V). PE :V EA =5:1), yielding an oily substance 38-2 (350 mg, yield 62.5%).

[0456] Step 2: Synthesis of compound 38-3

[0457] Dissolve 38-2 (350 mg, 0.875 mmol) in 5 mL of ethyl acetate, add 2.5 mL of dioxane hydrochloride solution, stir at room temperature for 4 hours, filter, and give white solid 38-3 (250 mg, yield 85%).

[0458] Step 3: Synthesis of compound 38-4

[0459] Dissolve 1-1 (200 mg, 0.266 mmol) and HATU (121.22 mg, 0.319 mmol) in THF (5 mL), then add DIEA (137 mg, 1.06 mmol). Stir the reaction at room temperature for 0.5 hours, then add 38-3 (107.2 mg, 0.319 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 38-4 (170 mg, yield 62%).

[0460] Step 4: Synthesis of Compound 38

[0461] Compound 38-4 (170 mg, 0.164 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. Target compound 38 was a white solid (110 mg, yield 73.8%), [M+H]. + :836.8.

[0462] 1 H NMR(400MHz,CD3OD)δ7.53-7.29(m,14H),4.73-4.71(m,2H),4.42-4.39(m,1H),4.16- 4.13(m,2H),3.90-3.85(m,1H),3.03-3.01(m,8H),1.73-1.63(m,13H),0.99(dd,6H).

[0463] Example 39

[0464] Synthesis of (R)-N-((R)-6-amino-1-(4-((E)-3-(3,4-dichlorophenyl)-N-methacrylamide)piperidin-1-yl)-1-oxohexane-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide (39)

[0465]

[0466] Step 1: Synthesis of Compound 39-2

[0467] Dissolve 39-1 (211 mg, 0.973 mmol), 6-1 (250 mg, 1.168 mmol), HOBT (157.7 mg, 1.168 mmol), and EDCI.HCl (279.9 mg, 1.46 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (98.5 mg, 0.973 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate aqueous solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude product 39-2 (398 mg, yield 99.3%).

[0468] Step 2: Synthesis of compound 39-3

[0469] Dissolve 39-2 (398 mg, 0.966 mmol) in 5 mL of ethyl acetate, add 2.5 mL of dioxane hydrochloride, stir at room temperature for 2 hours, filter, and give white solid 39-3 (341 mg, yield 98.1%).

[0470] Step 3: Synthesis of compound 39-4

[0471] Dissolve 1-1 (150 mg, 0.199 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 39-3 (73 mg, 0.239 mmol) and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 39-4 (130 mg, yield 63.4%).

[0472] Step 4: Synthesis of Compound 39

[0473] Compound 39-4 (130 mg, 0.087 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride solution was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. Target compound 39 was a white solid (50 mg, yield 47.2%), [M+H]. + :848.1.

[0474] 1 H NMR (400MHz, CD3OD) δ7.82-7.80(m,1H),7.58-7.53(m,2H),7.37-7.31(m,10H),4.73-4.71(m,2 H),4.42-4.39(m,2H),4.16-4.13(m,1H),,3.03-3.01(m,9H),1.73-1.63(m,13H),0.99(dd,6H).

[0475] Example 40

[0476] Synthesis of (R)-N-((R)-6-amino-1-(4-((E)-N-methyl-3-(4-(trifluoromethyl)phenyl)acrylamide)piperidin-1-yl)-1-oxohexane-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide hydrochloride (40)

[0477]

[0478] Step 1: Synthesis of Compound 40-2

[0479] Dissolve 40-1 (210 mg, 0.973 mmol), 6-1 (250 mg, 1.168 mmol), HOBT (157.7 mg, 1.168 mmol), and EDCI.HCl (277.9 mg, 1.46 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (98.5 mg, 0.973 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate aqueous solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude product 40-2 (340 mg, yield 84.9%).

[0480] Step 2: Synthesis of Compound 40-3

[0481] The crude product 40-2 (340 mg, 0.825 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride solution was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 40-3 (285 mg, yield 99.3%).

[0482] Step 3: Synthesis of compound 40-4

[0483] Dissolve 1-1 (150 mg, 0.199 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 40-3 (83.2 mg, 0.239 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 40-4 (130 mg, yield 63.4%).

[0484] Step 4: Synthesis of Compound 40

[0485] 40-4 (130 mg, 0.124 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. Target compound 40 was a brown solid (70 mg, yield 61.4%), [M+H]. + :848.14.

[0486] 1 H NMR (400MHz, CD3OD) δ7.90-7.81(m,2H),7.73-7.71(d,2H),7.63-7.60(m,1H),7.37-7.31(m,11H),4.73-4.71(m,3 H),4.42-4.39(m,1H),4.16-4.13(m,1H),3.28-3.24(m,1H),3.03-2.88(m,7H),1.73-1.63(m,13H),0.99(dd,6H).

[0487] Example 41

[0488] Synthesis of (R)-N-((R)-6-amino-1-(4-(2-(3,4-difluorophenyl)acetamyl)piperidin-1-yl)-1-oxohexane-2-yl)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide hydrochloride (41)

[0489] Step 1: Synthesis of Compound 41-2

[0490] Dissolve 41-1 (179.2 mg, 1.042 mmol), 7-1 (250 mg, 1.25 mmol), HOBT (168.7 mg, 1.25 mmol), and EDCI.HCl (299.5 mg, 1.56 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (105.2 mg, 1.042 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate aqueous solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude product 41-2 (295 mg, yield 80.1%).

[0491] Step 2: Synthesis of compound 41-3

[0492] The crude product 41-2 (295 mg, 0.833 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 41-3 (241 mg, yield 81.7%).

[0493] Step 3: Synthesis of compound 41-4

[0494] Dissolve 1-1 (150 mg, 0.199 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). After stirring at room temperature for 0.5 hours, add 41-3 (69 mg, 0.239 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 41-4.

[0495] Step 4: Synthesis of Compound 41

[0496] 41-4 (130 mg, 0.131 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. The target compound 41 was a brown solid (80 mg, 51% yield in two steps), [M+H] + 790.98.

[0497] 1H NMR (400MHz, CD3OD) δ7.37-7.07(m,13H),486-4.84(m,1H),4.75-4.70(m,1H),4.42-4.39(m,2 H),4.16-4.13(m,1H),3.28-3.24(m,2H),3.03-2.88(m,7H),1.73-1.63(m,13H),0.99(dd,6H).

[0498] Example 42 Synthesis of (R)-N-((R)-6-amino-1-(4-(N-methyl-3-phenylpropamido)piperidin-1-yl)-1-oxohexane-2-yl)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide hydrochloride (42)

[0499] Step 1: Synthesis of Compound 42-2

[0500] Dissolve 42-1 (142.1 mg, 0.973 mmol), 6-1 (250 mg, 1.168 mmol), HOBT (157.7 mg, 1.168 mmol), and EDCI.HCl (279.9 mg, 1.46 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (98.5 mg, 0.973 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate aqueous solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude product 42-2 (210 mg, yield 74.5%).

[0501] Step 2: Synthesis of compound 42-3

[0502] The crude product 42-2 (210 mg, 0.614 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 42-3 (167 mg, yield 97.5%).

[0503] Step 3: Synthesis of compound 42-4

[0504] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 42-3 (66.4 mg, 0.239 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH=10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 42-4 (170 mg, yield 43.7%).

[0505] Step 4: Synthesis of Compound 42

[0506] Compound 42-4 (170 mg, 0.174 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. Target compound 42 was a white solid (50 mg, yield 37%), [M+H]. + :778.2.

[0507] 1 H NMR (400MHz, CD3OD) δ7.57-7.25(m,15H),4.76-4.41(m,5H),3.34-3.24(m,2H),3.03-2.85(m,7H),1.73-1.63(m,13H),0.99(dd,6H).

[0508] Example 43

[0509] Synthesis of (R)-N-((R)-6-amino-1-(4-(3-(3,4-dichlorophenyl)-N-methylpropylamino)piperidin-1-yl)-1-oxo-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropionamide)-3-phenylpropionamide)-4-methylpentanamide hydrochloride (43)

[0510]

[0511] Step 1: Synthesis of Compound 43-2

[0512] Dissolve 43-1 (213.2 mg, 0.973 mmol), 6-1 (250 mg, 1.168 mmol), HOBT (157.7 mg, 1.168 mmol), and EDCI.HCl (279.9 mg, 1.46 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (98.5 mg, 0.973 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate aqueous solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude product 43-2 (291 mg, yield 72.1%).

[0513] Step 2: Synthesis of compound 43-3

[0514] The crude product 43-2 (291 mg, 0.703 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 43-3 (239 mg, yield 97.1%).

[0515] Step 3: Synthesis of compound 43-4

[0516] Dissolve 1-1 (150 mg, 0.199 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 43-3 (83 mg, 0.239 mmol) and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 43-4 (111 mg, yield 53.1%).

[0517] Step 4: Synthesis of Compound 43

[0518] Compound 43-4 (150 mg, 0.143 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. Target compound 43 was a brown solid (100 mg, yield 75.8%), [M+H]. + :850.84.

[0519] 1 H NMR(400MHz,CD3OD)δ7.37-7.07(m,13H),4.86-4.41(m,5H),4.15-4.13(m,2 H),3.34-3.32(m,2H),3.03-2.70(m,8H),1.73-1.63(m,13H),0.99(dd,6H).

[0520] Example 44

[0521] Synthesis of (R)-N-((R)-6-amino-1-oxo-1-(4-((E)-3-(pyridin-4-yl)acrylamide)piperidin-1-yl)hexane-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide hydrochloride (44)

[0522]

[0523] Step 1: Synthesis of Compound 44-2

[0524] Dissolve 44-1 (116.2 mg, 0.785 mmol), 6-1 (200 mg, 0.942 mmol), HOBT (127.2 mg, 0.942 mmol), and EDCI.HCl (224.0 mg, 1.169 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (78.8 mg, 0.785 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate aqueous solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude product 44-2 (201 mg, yield 74.8%).

[0525] Step 2: Synthesis of compound 44-3

[0526] The crude product 44-2 (201 mg, 0.583 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 44-3 (162 mg, yield 98.7%).

[0527] Step 3: Synthesis of compound 44-4

[0528] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 44-3 (75.8 mg, 0.239 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 44-4 (160 mg, yield 41.0%).

[0529] Step 4: Synthesis of Compound 44

[0530] Compound 44-4 (160 mg, 0.163 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride solution was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. Target compound 44 was a white solid (70 mg, yield 48.3%), [M+H]. + 782.06.

[0531] 1 H NMR(400MHz,CD3OD)δ8.87-8.85(d,2H),8.44-8.31(m,2H),8.01-7.91(m,1H),7.77-7.59(m,2H),7.36-7 .25(m,10H),4.76-4.41(m,5H),3.34-3.24(m,3H),3.03-2.85(m,8H),1.73-1.63(m,13H),0.99(dd,6H).

[0532] Example 45

[0533] Synthesis of 4-((E)-3-((1-((R)-6-amino-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanyl)hexanoyl)piperidin-4-yl)amino)-3-oxopropyl-1-en-1-yl)-1-benzylpyridine-1-onium hydrochloride (45)

[0534]

[0535] Step 1: Synthesis of Compound 45-2

[0536] Dissolve 44-1 (116.2 mg, 0.785 mmol), 7-1 (200 mg, 0.942 mmol), HOBT (127.2 mg, 0.942 mmol), and EDCI.HCl (224.0 mg, 1.169 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (78.8 mg, 0.785 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate aqueous solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude product 45-2 (200 mg, yield 77.6%).

[0537] Step 2: Synthesis of Compound 45-3

[0538] The crude product 45-2 (200 mg, 0.604 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours and then filtered to give a white solid 45-3 (145 mg, yield 89.6%).

[0539] Step 3: Synthesis of compound 45-4

[0540] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 45-3 (75.8 mg, 0.239 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 45-4.

[0541] Step 4: Synthesis of compound 45-5

[0542] Dissolve 45-4 (150 mg, 0.148 mmol) in 5 mL of dichloromethane, add benzyl bromide (27.8 mg, 0.162 mmol), stir overnight at room temperature, concentrate the solvent to dryness, wash with crude ethyl acetate, filter, and give white solid 45-5.

[0543] Step 5: Synthesis of Compound 45

[0544] The above solid 45-5 was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to obtain the product. Target compound 45 was a white solid (40 mg, two-step yield 30.0%), [M+H]. + :872.2.

[0545] 1 H NMR(400MHz,CD3OD)δ9.05-9.01(d,2H),8.46-8.33(m,2H),7.78-7.61(m,2H),7.56-7.49(m,5H),7.37-7.22(m,10 H),4.76-4.63(m,3H),4.49-4.38(m,1H),4.17-4.14(m,1H),3.15-2.92(m,8H),1.73-1.63(m,13H),0.99(dd,6H).

[0546] Example 46

[0547] Synthesis of (2R)-N-((2R)-6-amino-1-(5-cinnamoylhexahydropyrrole[3,4-c]pyrrole-2(1H)-yl)-1-oxohexadecane-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentylamine hydrochloride (46)

[0548] Step 1: Synthesis of Compound 46-2

[0549] Dissolve 32-1 (116.2 mg, 0.785 mmol), 46-1 (200 mg, 0.942 mmol), HOBT (127.2 mg, 0.942 mmol), and EDCI.HCl (224.0 mg, 1.169 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (79.3 mg, 0.785 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate aqueous solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude product 46-2 (219 mg, yield 81.7%).

[0550] Step 2: Synthesis of Compound 46-3

[0551] The crude product 46-2 (219 mg) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 46-3 (145 mg, yield 81.2%).

[0552] Step 3: Synthesis of compound 46-4

[0553] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 46-3 (66.4 mg, 0.239 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 46-4 (160 mg, yield 41.1%).

[0554] Step 4: Synthesis of Compound 46

[0555] Compound 46-4 (160 mg, 0.164 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. Target compound 46 was a white solid (70 mg, yield 50.4%), [M+H]. + :778.6.

[0556] 1 H NMR (400MHz, CD3OD) δ7.67-6.97(m,15H),4.76-4.66(m,1H),4.44-4.28(m,1H),4.06-3.99(m, 1H),3.90-3.75(m,2H),3.54-3.39(m,2H),3.22-2.90(m,8H),1.93-1.31(m,10H),0.99(dd,6H)

[0557] Example 47

[0558] Synthesis of N-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanyl)hexanoyl)piperidin-4-yl)-N-methyl-2-phenylcyclopropanecarboxamide hydrochloride (47)

[0559] Step 1: Synthesis of Compound 47-2

[0560] Dissolve 47-1 (126.4 mg, 0.785 mmol), 6-1 (200 mg, 0.942 mmol), HOBT (127.2 mg, 0.942 mmol), and EDCI.HCl (224.0 mg, 1.169 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (78.8 mg, 0.785 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate aqueous solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude product 47-2 (208 mg, yield 74.7%).

[0561] Step 2: Synthesis of Compound 47-3

[0562] The crude product 47-2 (208 mg) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 47-3 (152 mg, yield 88.7%).

[0563] Step 3: Synthesis of compound 47-4

[0564] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 47-3 (70 mg, 0.239 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 47-4 (160 mg, yield 40.5%).

[0565] Step 4: Synthesis of Compound 47

[0566] Compound 47-4 (160 mg, 0.161 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. Target compound 47 was a white solid (50 mg, yield 34%), [M+H]. + :794.08.

[0567] 1 H NMR(400MHz,CD3OD)δ7.37-7.16(m,10H),4.76-4.63(m,3H),4.49-4.38(m,1H),4.17- 4.14(m,1H),3.26-3.18(m,1H),3.15-2.92(m,8H),1.73-1.63(m,17H),0.99(dd,6H).

[0568] Example 48

[0569] Synthesis of (R)-N-((R)-6-amino-1-(4-(N-cyclopropylcinnamamido)piperidin-1-yl)-1-oxohexane-2-yl)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide hydrochloride (48)

[0570] Step 1: Synthesis of Compound 48-2

[0571] Dissolve 30-1 (523 mg, 2.63 mmol) in 10 mL of dichloromethane, add cyclopropylamine (100 mg, 1.75 mmol), stir at room temperature for 0.5 hours, add NaBH(AcO)3 (929 mg, 4.38 mmol), and stir for 3 hours. Quench the reaction with water, extract three times with dichloromethane, combine the organic phases, concentrate, and perform column chromatography (V). DCM :V MeOH =20:1), yielding product 48-2 (552 mg, yield 87.5%).

[0572] Step 2: Synthesis of Compound 48-3

[0573] Dissolve 48-2 (130 mg, 0.542 mmol), 32-1 (66.7 mg, 0.451 mmol), HOBT (73.1 mg, 0.542 mmol), and EDCI.HCl (129.8 mg, 0.677 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (45.56 mg, 0.451 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate aqueous solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude product 48-3 (211 mg, yield 64.9%).

[0574] Step 3: Synthesis of compound 48-4

[0575] The crude product 48-3 (211 mg, 0.570 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 48-4 (152 mg, yield 86.9%).

[0576] Step 4: Synthesis of compound 48-5

[0577] Dissolve 1-1 (102 mg, 0.135 mmol) and HATU (62 mg, 0.162 mmol) in THF (5 mL), then add DIEA (70.4 mg, 0.542 mmol). Stir the reaction at room temperature for 0.5 hours, then add 48-4 (50 mg, 0.163 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 48-5 (60mg, yield 44.1%).

[0578] Step 5: Synthesis of Compound 48

[0579] 48-5 (60 mg, 0.060 mmol) was dissolved in 3 mL of ethyl acetate, and then 1.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. Target compound 48 was a white solid (30 mg, yield 57.7%).

[0580] [M+H] + :805.14; 1 H NMR (400MHz, CD3OD) δ7.67-7.31(m,15H),4.73-4.45(m,2H),4.42-4.39(m,1H),4.16-4.13(m,2 H),3.28-3.24(m,1H),3.03-2.88(m,7H),1.73-1.63(m,13H),0.99(dd,6H),0.89-0.77(m,4H).

[0581] Example 49

[0582] Synthesis of (R)-N-((R)-6-amino-1-oxo-1-(4-(2-oxo-2,3-dihydro-1H-imidazo[4,5-b]pyridin-1-yl)piperidin-1-yl)hexan-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide hydrochloride (49)

[0583]

[0584] Step 1: Synthesis of Compound 49-2

[0585] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 49-1 (75.8 mg, 0.239 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 49-2 (120 mg, yield 31.6%).

[0586] Step 2: Synthesis of Compound 49

[0587] Compound 49-2 (120 mg, 0.126 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. Target compound 49 was a white solid (90 mg, yield 86.6%), [M+H]. + :754.9.

[0588] 1 H NMR(400MHz,CD3OD)δ8.18-7.91(m,3H),7.41-7.21(m,10H),4.76-4.65(m, 3H),4.41-4.21(m,2H),3.03-2.85(m,8H),1.73-1.63(m,13H),0.99(dd,6H)

[0589] Example 50

[0590] Synthesis of (R)-N-((R)-6-amino-1-(4-(3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-b]pyridin-1-yl)piperidin-1-yl)-1-oxohexane-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamine)-3-phenylpropylamine)-4-methylpentanamide hydrochloride (50)

[0591]

[0592] Step 1: Synthesis of Compound 50-2

[0593] Dissolve 49-1 (50 mg, 0.229 mmol) in 2 mL of dichloromethane, add 23 mL of triethylamine and (Boc)2O (74.8 mg, 0.343 mmol), stir at room temperature for 1.5 hours, quench the reaction with water, extract three times with dichloromethane, concentrate, and give a yellow solid. Beat with n-hexane to give product 50-2 (73 mg, yield 99.9%).

[0594] Step 2: Synthesis of Compound 50-3

[0595] The above product 50-2 (73 mg, 0.229 mmol) was dissolved in 2 mL of THF, and then sodium hydride (6 mg, 0.25 mmol) and methyl iodoform (35.8 mg, 0.252 mmol) were added. The mixture was stirred at room temperature for 4 hours, and the reaction was quenched with water. DCM :V MeOH Extracted three times at a ratio of 10:1, concentrated to obtain product 50-3 (50mg, yield 65.6%).

[0596] Step 3: Synthesis of Compound 50-4

[0597] The above product was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours and filtered to give a white solid product 50-4 (40 mg, yield 98.8%).

[0598] Step 4: Synthesis of Compound 50

[0599] Dissolve 1-1 (187 mg, 0.248 mmol) and HATU (57 mg, 0.149 mmol) in THF, add DIEA (64 mg, 0.496 mmol), and stir at room temperature for 0.5 hours. Then add 50-4 (40 mg, 0.149 mmol), and stir at room temperature overnight. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and use V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding 50mg of product.

[0600] The above product was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride solution was added. The mixture was stirred at room temperature for 2 hours, filtered, and the product was obtained. The target compound was a white solid (30 mg, two-step yield 14.4%), [M+H]. + :767.5.

[0601] 1 H NMR(400MHz,CD3OD)δ8.19-8.09(m,2H),7.51-7.24(m,13H),4.75-4.63(m,4H),4.49 -4.43(m,1H),3.26-3.23(m,2H),2.99-2.17(m,7H),1.89-1.4(m,13H),0.99(dd,6H)

[0602] Example 51

[0603] Synthesis of N1-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanyl)hexanoyl)piperidin-4-yl)-N2-methyloxalamide hydrochloride (51)

[0604]

[0605] Step 1: Synthesis of Compound 51-1

[0606] Dissolve 37-2 (330 mg, 1.154 mmol) in 2 mL of THF, add 1 mL of saturated sodium hydroxide solution, stir at room temperature for 3 hours, concentrate the THF to dryness, extract the aqueous phase three times with dichloromethane, combine the organic phases, adjust the pH to acidity with 2N hydrochloric acid, and dry with anhydrous sodium sulfate to obtain 310 mg of white solid.

[0607] The above-mentioned white solid (310 mg, 1.139 mmol), methylamine hydrochloride (92.4 mg, 1.367 mmol), HOBT (184.5 mg, 1.367 mmol), and EDCI.HCl (327.6 mg, 1.709 mmol) were dissolved in 5 mL of dichloromethane. After stirring at room temperature for 5 min, triethylamine (230.08 mg, 2.278 mmol) was added, and the mixture was stirred at room temperature overnight. The reaction was quenched with saturated sodium bicarbonate aqueous solution, and the organic phase was separated. The aqueous phase was extracted three times with dichloromethane. The combined organic phases were washed with 1N dilute hydrochloric acid, concentrated, and subjected to column chromatography (V). DCM :V MeOH =20:1), yielding 130 mg of product. The above product 130 mg was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride solution was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give white solid 51-1 (99 mg, yield 97.9%).

[0608] Step 2: Synthesis of compound 51-2

[0609] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 51-1 (52.9 mg, 0.239 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 51-2.

[0610] Step 4: Synthesis of Compound 51

[0611] The above product 51-2 was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to obtain the product. Target compound 51 was a white solid (50 mg, 16% yield in two steps), [M+H] + :721.

[0612] 1H NMRδ7.32-7.23(m,10H),4.81-4.71(m,2H),4.51-4.39(m,2H),4.20-4.00(m, 2H),3.26-3.18(m,1H),3.15-2.92(m,11H),1.73-1.63(m,13H),0.99(dd,6H).

[0613] Example 52

[0614] Synthesis of 1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanyl)hexanoyl)-N-(benzofuran-5-yl)piperidine-4-carboxamide hydrochloride (52)

[0615]

[0616] Step 1: Synthesis of Compound 52-2

[0617] Dissolve 52-1a (286.98 mg, 1.251 mmol), 52-1b (200 mg, 1.502 mmol), HOBT (202.93 mg, 1.502 mmol), and EDCI.HCl (359.7 mg, 1.876 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (126.35 mg, 1.251 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate aqueous solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude product 52-2 (401 mg, yield 93.0%).

[0618] Step 2: Synthesis of compound 52-3

[0619] The crude product 52-2 (401 mg) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 52-3 (295 mg, yield 90.2%).

[0620] Step 3: Synthesis of compound 52-4

[0621] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 52-3 (66.99 mg, 0.239 mmol) and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and use V...DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 52-4 (150 mg, yield 38.5%).

[0622] Step 4: Synthesis of Compound 52

[0623] Compound 52-4 (150 mg, 0.153 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. The target compound 52 was a white solid (70 mg, yield 53.8%), [M+H]. + 780.04.

[0624] 1 H NMR(400MHz,CD3OD)δ7.89(d,1H),7.73(d,1H),7.43-7.20(m,13H),4.41-4.35(m,4H),4. 13-4.07(m,1H),3.33-3.18(m,2H),3.15-2.92(m,8H),1.73-1.63(m,13H),0.99(dd,6H).

[0625] Example 53

[0626] Synthesis of (R)-N-((R)-6-amino-1-(4-(2-(benzofuran-5-ylamino)-2-oxoethyl)piperidin-1-yl)-1-oxohexane-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide hydrochloride (53)

[0627]

[0628] Step 1: Synthesis of Compound 53-2

[0629] Dissolve 53-1 (304.3 mg, 1.251 mmol), 52-1b (200 mg, 1.502 mmol), HOBT (202.93 mg, 1.502 mmol), and EDCI.HCl (359.7 mg, 1.876 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (126.35 mg, 1.251 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate aqueous solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude product 53-2 (424 mg, yield 94.7%).

[0630] Step 2: Synthesis of compound 53-3

[0631] The crude product 53-2 (424 mg, 1.184 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride solution was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 53-3 (316 mg, yield 90.5%).

[0632] Step 3: Synthesis of compound 53-4

[0633] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 53-3 (70.4 mg, 0.239 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 53-4 (180 mg, yield 45.5%).

[0634] Step 4: Synthesis of Compound 53

[0635] Dissolve 53-4 (180 mg, 1.181 mmol) in 5 mL of ethyl acetate, add 2.5 mL of dioxane hydrochloride, stir at room temperature for 2 hours, filter, and give the product. The target compound is a brown solid (90 mg, yield 57.3%), [M+H]. + :795.04.

[0636] 1 H NMR(400MHz,CD3OD)δ7.89(d,1H),7.73(d,1H),7.43-7.20(m,13H),4.41-4.35(m,4H),4. 13-4.07(m,1H),3.33-3.18(m,2H),3.15-2.92(m,8H),1.73-1.63(m,15H),0.99(dd,6H).

[0637] Example 54

[0638] Synthesis of (R)-N-((R)-6-amino-1-oxy-1-(4-(2-oxazolazole[d]oxazol-3(2H)-yl)piperidin-1-yl)hex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropionamide)-3-phenylpropionamide)-4-methylpentanamide hydrochloride (54)

[0639] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 54-1 (52 mg, 0.239 mmol) and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding the product.

[0640] The above product was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to obtain the product. Target compound 54 was a brown solid (50 mg, two-step yield 15.2%), [M+H]. + :754.94.

[0641] 1 H NMR(400MHz,CD3OD)δ7.30-7.20(m,10H),4.75-4.71(m,2H),4.45-4.43(m,1H),4.27- 4.13(m,2H),3.21-3.18(m,2H),2.99-2.17(m,9H),1.73-1.63(m,13H),0.99(dd,6H).

[0642] Example 55

[0643] Synthesis of N-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanyl)hexanoyl)piperidin-4-yl)benzofuran-2-carboxamide hydrochloride (55)

[0644]

[0645] Step 1: Synthesis of Compound 55-2

[0646] Dissolve 35-1 (168.5 mg, 1.04 mmol), 7-1 (250 mg, 1.25 mmol), HOBT (168.7 mg, 1.25 mmol), and EDCI.HCl (299.5 mg, 1.56 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (105.2 mg, 1.04 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate aqueous solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude product 55-2 (314 mg, yield 87.5%).

[0647] Step 2: Synthesis of Compound 55-3

[0648] The crude product 55-2 (314 mg) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 55-3 (248 mg, yield 96.8%).

[0649] Step 3: Synthesis of compound 55-4

[0650] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 55-3 (67 mg, 0.239 mmol) and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 55-4 (110 mg, yield 28.2%).

[0651] Step 4: Synthesis of Compound 55

[0652] Compound 55-4 (180 mg, 0.184 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. The target compound 55 was a white solid (70 mg, 44.9%), [M+H]. + :780.4.

[0653] 1H NMR (400MHz, CD3OD) δ8.50-8.48(m,1H),7.80-7.71(m,1H),7.63-7.18(m,14H),4.75-4.40(m, 3H),4.25-4.11(m,2H),3.26-3.23(m,2H),2.99-2.81(m,5H),1.89-1.4(m,13H),0.99(dd,6H).

[0654] Example 56

[0655] Synthesis of 2-(8-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanyl)hexyl)-3,8-diazabicyclo[3.2.1]octane-3-yl)-2-oxoacetic acid hydrochloride (56)

[0656] Step 1: Synthesis of Compound 56-2

[0657] 56-1 (300 mg, 1.42 mmol), 37-1 (190.7 mg, 1.56 mmol), and triethylamine (172.1 mg, 1.7 mmol) were dissolved in 5 mL of dichloromethane, stirred at room temperature for 2 hours, the reaction was quenched with water, and the mixture was extracted three times with dichloromethane. The organic phases were combined and concentrated to give crude product 2 (405 mg, yield 87.8%).

[0658] Step 2: Synthesis of Compound 56-3

[0659] The crude product 56-2 (405 mg, 1.359 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 56-3 (349 mg, yield 86.2%).

[0660] Step 3: Synthesis of compound 56-4

[0661] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 56-3 (25 mg, 0.239 mmol) and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction 3 times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH=20:1), yielding product 56-4 (120 mg, yield 32.3%).

[0662] Step 4: Synthesis of compound 56-5

[0663] The product 56-4 (120 mg, 1.128 mmol) was dissolved in 5 mL of THF, and then 2 mL of saturated NaOH aqueous solution was added. The mixture was stirred at room temperature for 3 hours. The THF was concentrated to dryness, and the aqueous phase was extracted three times with dichloromethane. The organic phases were combined, the pH was adjusted to acidity with 1 N hydrochloric acid, and the mixture was concentrated to give product 56-5 (100 mg, yield 84.6%).

[0664] Step 5: Synthesis of Compound 56

[0665] The above product was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 3 hours, and then filtered to obtain the product. Target compound 56 was a brown solid (30 mg, yield 34.8%), [M+H]. + :720.8.

[0666] 1 H NMR (400MHz, CD3OD) δ7.36-7.31(m,10H),4.86-4.85(m,1H),4.75-4.72(m,2H),4.60-4.58(m,1H),4.46- 4.39(m,2H),4.15-4.09(m,1H),3.26-3.23(m,1H),2.99-2.81(m,6H),1.89-1.47(m,13H),0.99(dd,6H).

[0667] Example 57

[0668] Synthesis of ethyl 2-(4-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanyl)hexanoyl)piperazin-1-yl)-2-oxoacetic acid hydrochloride (57)

[0669]

[0670] Step 1: Synthesis of Compound 57-2

[0671] 56-1 (300 mg, 1.42 mmol), 57-1 (167.5 mg, 1.56 mmol), and triethylamine (172.1 mg, 1.7 mmol) were dissolved in 5 mL of dichloromethane, stirred at room temperature for 2 hours, the reaction was quenched with water, and the mixture was extracted three times with dichloromethane. The organic phases were combined and concentrated to obtain crude product 57-2 (405 mg, yield 87.8%).

[0672] Step 2: Synthesis of Compound 57-3

[0673] The crude product 57-2 (405 mg, 1.30 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 57-3 (349 mg, yield 86.2%).

[0674] Step 3: Synthesis of compound 57-4

[0675] Dissolve 1-1 (400 mg, 0.531 mmol) and HATU (121 mg, 0.319 mmol) in THF (5 mL), then add DIEA (137 mg, 1.06 mmol). Stir the reaction at room temperature for 0.5 hours, then add 57-3 (74 mg, 0.319 mmol) and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction three times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 57-4.

[0676] Step 4: Synthesis of Compound 57

[0677] The product 57-4 was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to obtain the product. The target compound 57 was a brown solid (40 mg, 10% yield in two steps), [M+H]. + :722.66.

[0678] 1 H NMR (400MHz, CD3OD) δ7.39-7.22(m,10H),4.75-4.72(m,1H),4.59-4.37(m,2H),4.13-4.11(m, 1H),3.92-3.89(m,1H),3.26-3.23(m,1H),3.04-2.89(m,8H),1.89-1.47(m,7H),0.99(dd,6H).

[0679] Example 58

[0680] Synthesis of N-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanyl)hexanoyl)-4-methylpiperidin-4-yl)benzofuran-6-carboxamide hydrochloride (58)

[0681] Step 1: Synthesis of Compound 58-2

[0682] Dissolve 24-1 (200 mg, 0.92 mmol), 34-1 (126.4 mg, 0.78 mmol), HOBT (126.2 mg, 0.93 mmol), and EDCI.HCl (224 mg, 1.17 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (79 mg, 0.78 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate aqueous solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude product 58-2 (314 mg, yield 93.8%).

[0683] Step 2: Synthesis of Compound 58-3

[0684] The crude product 58-2 (314 mg, 0.877 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride solution was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 58-3 (243 mg, yield 94.0%).

[0685] Step 3: Synthesis of compound 58-4

[0686] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 58-3 (70.3 mg, 0.239 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction three times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 58-4.

[0687] Step 4: Synthesis of Compound 58

[0688] The above product 58-4 was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to obtain the product. The target compound was a brown solid (50 mg, two-step yield 14.5%), [M+H]. + 794.98.

[0689] 1H NMR (400MHz, CD3OD) δ7.59-7.48(m,3H),7.41-7.23(m,13H),6.79-6.72(m,1H),4.77-4.70(m,1H),4.46-4.38(m,1 H),4.16-4.11(m,1H),3.46-3.40(m,1H),3.28-3.19(m,2H),3.04-2.89(m,8H),1.89-1.47(m,13H),0.99(dd,6H).

[0690] Example 59

[0691] Synthesis of (R)-N-((R)-6-amino-1-(4-(2-(3,4-dichlorophenyl)acetamyl)-4-methylpiperidin-1-yl)-1-oxohexane-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide hydrochloride (59)

[0692]

[0693] Step 1: Synthesis of Compound 59-2

[0694] Dissolve 24-1 (250 mg, 1.17 mmol), 59-1 (200 mg, 0.98 mmol), HOBT (158 mg, 1.17 mmol), and EDCI.HCl (282 mg, 1.47 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (100 mg, 0.98 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude 59-2 (389 mg, yield 83.0%).

[0695] Step 2: Synthesis of Compound 59-3

[0696] The crude product 59-2 (389 mg, 0.972 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 59-3 (301 mg, yield 91.9%).

[0697] Step 3: Synthesis of compound 59-4

[0698] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 59-3 (80.5 mg, 0.239 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction three times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 59-4.

[0699] Step 4: Synthesis of Compound 59

[0700] The above product 59-4 was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to obtain the product. The target compound was a brown solid (80 mg, two-step yield 22.2%), [M+H]. + :836.8.

[0701] 1 H NMR (400MHz, CD3OD) δ7.55-7.24(m,13H),4.89-4.86(m,1H),4.75-4.72(m,1H),4.44-4.38(m,1H),4.16- 4.11(m,1H),3.53-3.50(m,2H),3.25-3.20(m,1H),3.04-2.89(m,7H),1.79-1.47(m,13H),0.99(dd,6H).

[0702] Example 60

[0703] Synthesis of (R)-N-((R)-6-amino-1-(4-cinnamamide-4-methylpiperidin-1-yl)-1-oxo-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropionamide)-3-phenylpropionamide)-4-methylpentanamide hydrochloride (60)

[0704] Step 1: Synthesis of Compound 60-2

[0705] Dissolve 24-1 (150 mg, 1.17 mmol), 32-1 (88 mg, 0.98 mmol), HOBT (95 mg, 1.17 mmol), and EDCI.HCl (169 mg, 1.47 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (60 mg, 0.98 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate aqueous solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, and concentrate to give crude product 60-2 (214 mg, yield 88.8%).

[0706] Step 2: Synthesis of Compound 60-3

[0707] The crude product 60-2 (214 mg, 0.622 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours and then filtered to obtain a white solid 60-3 (171 mg, yield 97.9%).

[0708] Step 3: Synthesis of Compound 60-4

[0709] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 60-3 (67 mg, 0.239 mmol) and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction three times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 60-4.

[0710] Step 4: Synthesis of Compound 60

[0711] The above product 60-4 (125 mg, 0.13 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to obtain the product. The target compound was a brown solid (70 mg, two-step yield 20.65%), [M+H]. + 780.86.

[0712] 1H NMR(400MHz,CD3OD)δ8.13-8.08(m,1H),7.88-7.85(m,1H),7.79-7.73(m,1 H),7.60-7.52(m,1H),7.39-7.24(m,10H),6.99-6.92(m,1H),4.75-4.65(m, 1H),4.44-4.38(m,1H),4.16-4.11(m,1H),3.56-3.50(m,1H),3.28-3.19(m ,1H),3.04-2.89(m,7H),2.04-1.85(t,3H)1.89-1.47(m,13H),0.99(dd,6H)

[0713] Example 61

[0714] Synthesis of (R)-N-((R)-6-amino-1-(2-methyl-3-oxo-2,8-diazaspiro[4.5]dec-8-yl)-1-oxohexane-2-yl)-4-methyl-2-((R)-3-phenyl-2-(2-((R)-2-phenylpropyl)amino)acetamide)propylamine)pentylamine trifluoroacetate (61)

[0715]

[0716] Step 1: Synthesis of Compound 61-2

[0717] Dissolve 8-2 (1.55 g, 8.13 mmol) in 15 mL of water, add Na2CO3 (2.15 g, 20.32 mmol) and THF (15 mL), and add (Boc)2O (2.13 g, 9.76 mmol) dropwise under ice bath. Stir the reaction at room temperature for 3 hours. Quench the reaction with water, extract three times with ethyl acetate, combine the organic phases, concentrate, and give a white solid 61-2 (1.68 g, yield 81%).

[0718] Step 2: Synthesis of compound 61-3

[0719] Dissolve 61-2 (150 mg, 0.59 mmol) in 5 mL THF, then add NaH (15 mg, 0.59 mmol) and MeI (90 mg, 0.65 mmol), and stir overnight at room temperature. Quench the reaction with water, extract three times with ethyl acetate, combine the organic phases, concentrate, and perform column chromatography (V... DCM :V MeOH =10:1), to obtain colorless oily substance 61-3 (90mg, yield 55.4%).

[0720] Step 3: Synthesis of compound 61-4

[0721] The above-mentioned oily substance 61-3 (90 mg) was dissolved in 3 mL of dichloromethane, and then 0.75 mL of trifluoroacetic acid was added. The mixture was stirred at room temperature for 2 hours, concentrated, and precipitated by adding diethyl ether. The solid was then filtered to obtain a white powder 61-4 (93 mg, yield 98.2%).

[0722] Step 4: Synthesis of compound 61-5

[0723] Dissolve 27-1 (150 mg, 0.192 mmol) and HATU (87 mg, 0.23 mmol) in 5 mL of THF, then add DIEA (99 mg, 0.768 mmol). Stir the reaction at room temperature for 0.5 hours, then add 61-4 (65 mg, 0.23 mmol) and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, extract the aqueous phase three times with ethyl acetate, combine the organic phases, concentrate, and perform column chromatography (V... DCM :V MeOH =20:1), yielding product 61-5 (100 mg, yield 56.2%).

[0724] Step 5: Synthesis of Compound 61

[0725] Dissolve 61-5 (100 mg) in 5 mL of dichloromethane, add 1 mL of trifluoroacetic acid, stir at room temperature for 2 hours, concentrate, add diethyl ether to precipitate the solid, filter, and give the product. The target compound is a brown powder (50 mg, yield 48.5%), [M+H] + :732.4.

[0726] 1 H NMR(400MHz,CD3OD)δ7.41-7.18(m,10H),4.75-4.19(m,1H),4.40-4.39(m,1H),3.92 -3.48(m,6H),3.21-2.83(m,9H),2.39-2.34(m,2H),1.78-1.38(m,16H),0.94(d,6H).

[0727] Example 62

[0728] Synthesis of 4-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamino)hexylamino)cyclohexanecarboxylate hydrochloride (62)

[0729]

[0730] Step 1: Synthesis of Compound 62-2

[0731] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir at room temperature for 0.5 hours, then add 62-1 (68.3 mg, 0.478 mmol), and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction three times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 62-2.

[0732] Step 2: Synthesis of Compound 62

[0733] The above product was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to obtain the product. The target compound was a brown solid 62 (80 mg, two-step yield 26.8%), [M+H]. + :679.4.

[0734] Example 63

[0735]

[0736] Step 1: Synthesis of Compound 63-2

[0737] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 63-1 (107.14 mg, 0.478 mmol) and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and use V... DCM :V MeOH =10:1 extraction three times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 63-2.

[0738] Step 2: Synthesis of Compound 63

[0739] The above product was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to obtain the product. The target compound was a brown solid 63 (80 mg, two-step yield 26.8%), [M+H]. + :756.44.

[0740] Example 64

[0741] Synthesis of methyl(2-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanylamino)-N-methylhexylamino)ethyl)-D-proline hydrochloride (64)

[0742]

[0743] Step 1: Synthesis of Compound 64-2

[0744] Dissolve 1-1 (300 mg, 0.398 mmol) and HATU (90.8 mg, 0.239 mmol) in THF (5 mL), then add DIEA (102.8 mg, 0.797 mmol). Stir the reaction at room temperature for 0.5 hours, then add 64-1 (107.14 mg, 0.478 mmol) and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and use V... DCM :V MeOH =10:1 extraction three times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 64-2.

[0745] Step 2: Synthesis of Compound 64

[0746] The above product was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to obtain the product. The target compound was a brown solid 64 (80 mg, two-step yield 26.8%), [M+H]. + :722.44.

[0747] Example 65

[0748] Synthesis of (R)-6-amino-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamino)-N-(4-(2-(3,4-difluorophenyl)-N-methylacetamido)cyclohexyl)-N-methylhexylamine hydrochloride (65)

[0749] Step 1: Synthesis of Compound 65-2

[0750] Dissolve 41-1 (2 g, 9.76 mmol), 6-1 (1.79 g, 11.71 mmol), HOBT (1.58 g, 11.71 mmol), and EDCI.HCl (2.8 g, 14.64 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (0.99 g, 9.76 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, concentrate, and perform column chromatography (V). PE :V EA =1:1), yielding an oily substance 65-2 (1.2g, yield 42%).

[0751] Step 2: Synthesis of Compound 65-3

[0752] Dissolve 65-2 (1.2 g, 4.21 mmol) in 20 mL of EA, add dioxane hydrochloride (4 M, 5 mL), stir for 2 hours, and then concentrate to obtain crude 65-3 (0.99 g, yield 99.6%).

[0753] Step 3: Synthesis of Compound 65-4

[0754] Dissolve 1-1 (500 mg, 0.664 mmol) and HATU (317 mg, 0.797 mmol) in THF (10 mL), then add DIEA (269 mg, 1.992 mmol). Stir the reaction at room temperature for 0.5 hours, then add 65-3 (250 mg, 0.797 mmol), and stir the reaction at room temperature for 7 hours. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction three times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =50:1), to obtain a white solid 65-4 (400 mg, yield 46%).

[0755] Step 4: Synthesis of Compound 65

[0756] Dissolve 65-4 (400 mg, 0.386 mmol) in 10 mL of ethyl acetate, add 5 mL of dioxane hydrochloride solution, stir at room temperature for 1.5 hours, filter, and give the product product, a white solid 65 (150 mg, yield: 83%), [M+H]. + :832.49.

[0757] Example 66

[0758] Synthesis of (R)-6-amino-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamino)-N-methyl-N-(4-((methyl-d3)((E)-styryl)amino)cyclohexyl)hexylamine hydrochloride (66)

[0759] Step 1: Synthesis of Compound 66-2

[0760] Dissolve 32-1 (189.3 mg, 1.128 mmol), 66-1 (300 mg, 1.38 mmol), HOBT (189 mg, 1.40 mmol), and EDCI.HCl (335.9 mg, 1.75 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (118.2 mg, 1.168 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, concentrate, and perform column chromatography (V). PE :V EA =5:1), yielding an oily substance 66-2 (360mg, yield 72%).

[0761] Step 2: Synthesis of Compound 66-3

[0762] Dissolve 66-2 (360 mg) in 5 mL of ethyl acetate, add 2.5 mL of dioxane hydrochloride, stir at room temperature for 4 hours, filter, and give white solid 66-3 (271 mg, yield 92.0%).

[0763] Step 3: Synthesis of Compound 66-4

[0764] Dissolve 1-1 (200 mg, 0.266 mmol) and HATU (121.22 mg, 0.319 mmol) in THF (5 mL), then add DIEA (137 mg, 1.06 mmol), stir at room temperature for 0.5 hours, then add 66-3 (93.79 mg, 0.428 mmol), stir at room temperature overnight. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and use V... DCM :V MeOH =10:1 extraction three times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 66-4 (180 mg, yield 66%).

[0765] Step 4: Synthesis of Compound 66

[0766] Dissolve 66-4 (180 mg, 0.184 mmol) in 5 mL of ethyl acetate, add 2.5 mL of dioxane hydrochloride, stir at room temperature for 2 hours, and filter to give the product. The target compound is a white solid 66 (100 mg, yield 66.2%), [M+H]. + :783.5.

[0767] Example 67

[0768] Synthesis of (R)-N-((R)-6-amino-1-oxo-1-(4-((E)-styryl)amino)piperidin-1-yl)hex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide hydrochloride (67)

[0769] Step 1: Synthesis of Compound 67-2

[0770] Dissolve 32-1 (189.3 mg, 1.128 mmol), 7-1 (300 mg, 1.38 mmol), HOBT (189 mg, 1.40 mmol), and EDCI.HCl (335.9 mg, 1.75 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (118.2 mg, 1.168 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, extract the aqueous phase three times with dichloromethane, combine the organic phases, wash with 1N dilute hydrochloric acid, concentrate, and perform column chromatography (V). PE :V EA =5:1), yielding an oily substance 67-2 (360 mg, yield 72%).

[0771] Step 2: Synthesis of Compound 67-3

[0772] Dissolve 67-2 (360 mg) in 5 mL of ethyl acetate, add 2.5 mL of dioxane hydrochloride, stir at room temperature for 4 hours, filter, and give white solid 67-3 (271 mg, yield 93.0%).

[0773] Step 3: Synthesis of Compound 67-4

[0774] Dissolve 1-1 (200 mg, 0.266 mmol) and HATU (121.22 mg, 0.319 mmol) in THF (5 mL), then add DIEA (137 mg, 1.06 mmol). Stir the reaction at room temperature for 0.5 hours, then add 67-3 (93.79 mg, 0.428 mmol), and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and use V... DCM :VMeOH =10:1 extraction three times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 67-4 (180 mg, yield 66%).

[0775] Step 4: Synthesis of Compound 67

[0776] 67-4 (180 mg, 0.186 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. The target compound was a white solid 67 (100 mg, yield 66.2%), [M+H]. + 766.5.

[0777] Example 68

[0778] Synthesis of (R)-N-((R)-6-amino-1-(4-(N-(cyclopropylmethyl)cinnamamido)piperidin-1-yl)-1-oxohexane-2-yl)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide hydrochloride (68)

[0779] Step 1: Synthesis of Compound 68-3

[0780] Dissolve 67-2 (200 mg, 0.61 mmol) in 5 mL THF, add NaH (48 mg, 1.22 mmol), stir at room temperature for 10 min, then add cyclopropyl bromide (163 mg, 1.22 mmol), and stir the reaction overnight at room temperature. Quench the reaction with water, extract three times with ethyl acetate, combine the organic phases, concentrate, and column chromatography (V... PE :V EA =3:1), yielding a pale yellow solid 68-3 (102 mg, yield 43.8%).

[0781] Step 2: Synthesis of Compound 68-4

[0782] Dissolve 68-3 (102 mg, 0.265 mmol) in 5 mL of ethyl acetate, add 2.5 mL of dioxane hydrochloride, stir at room temperature for 4 hours, filter, and give a white solid 68-4 (79 mg, yield 92.7%).

[0783] Step 3: Synthesis of Compound 68-5

[0784] Dissolve 1-1 (200 mg, 0.266 mmol) and HATU (121.22 mg, 0.319 mmol) in THF (5 mL), then add DIEA (137 mg, 1.06 mmol), stir at room temperature for 0.5 hours, then add 68-4 (84.8 mg, 0.265 mmol), and stir at room temperature overnight. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and use V... DCM :V MeOH =10:1 extraction three times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 68-5 (180 mg, yield 66%).

[0785] Step 4: Synthesis of Compound 68

[0786] Compound 68-5 (180 mg, 0.176 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. The target compound 68 was a white solid (100 mg, yield 69.4%), [M+H]. + :820.5.

[0787] Example 69

[0788] Synthesis of (R)-N-((R)-6-amino-1-(4-((S)-2-(2-fluoro-[1,1'-biphenyl]-4-yl)-N-methylpropylamino)piperidin-1-yl)-1-oxohexane-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamine hydrochloride (69)

[0789]

[0790] Step 1: Synthesis of Compound 69-2

[0791] Dissolve 69-1 (244 mg, 1 mmol), 6-1 (200 mg, 1 mmol), HOBT (189 mg, 1.40 mmol), and EDCI.HCl (335.9 mg, 1.75 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (118.2 mg, 1.168 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, concentrate, and perform column chromatography (V). PE :V EA =5:1), yielding an oily substance 69-2 (390 mg, yield 72%).

[0792] Step 2: Synthesis of Compound 69-3

[0793] Dissolve 69-2 (390 mg, 0.886 mmol) in 5 mL of ethyl acetate, add 2.5 mL of dioxane hydrochloride, stir at room temperature for 4 hours, filter, and give white solid 69-3 (327 mg, yield 97.9%).

[0794] Step 3: Synthesis of Compound 69-4

[0795] Dissolve 1-1 (200 mg, 0.266 mmol) and HATU (121.22 mg, 0.319 mmol) in THF (5 mL), then add DIEA (137 mg, 1.06 mmol). Stir the reaction at room temperature for 0.5 hours, then add 69-3 (161 mg, 0.428 mmol) and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction three times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 69-4 (280 mg, yield 55.94%).

[0796] Step 4: Synthesis of Compound 69

[0797] Dissolve 69-4 (280 mg, 0.260 mmol) in 5 mL of ethyl acetate, add 2.5 mL of dioxane hydrochloride, stir at room temperature for 2 hours, and filter to give the product. The target compound is a white solid 69 (210 mg, yield 91.7%), [M+H]. + :876.5.

[0798] Example 70

[0799] Synthesis of (R)-N-((R)-6-amino-1-(4-((S)-2-(6-methoxynaphthyl-2-yl)-N-methylpropylamino)piperidin-1-yl)-1-oxohexane-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamine hydrochloride (70)

[0800]

[0801] Step 1: Synthesis of Compound 70-2

[0802] Dissolve 70-1 (230 mg, 1 mmol), 6-1 (200 mg, 1 mmol), HOBT (189 mg, 1.40 mmol), and EDCI.HCl (335.9 mg, 1.75 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (118.2 mg, 1.168 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, concentrate, and perform column chromatography (V). PE :V EA =5:1), yielding 70-2 (390 mg, yield 72%) of oil.

[0803] Step 2: Synthesis of Compound 70-3

[0804] Dissolve 70-2 (390 mg, 0.886 mmol) in 5 mL of ethyl acetate, add 2.5 mL of dioxane hydrochloride, stir at room temperature for 4 hours, filter, and give white solid 70-3 (310 mg, yield 93.5%).

[0805] Step 3: Synthesis of compound 70-4

[0806] Dissolve 1-1 (200 mg, 0.266 mmol) and HATU (121.22 mg, 0.319 mmol) in THF (5 mL), then add DIEA (137 mg, 1.06 mmol), stir at room temperature for 0.5 hours, then add 70-3 (155 mg, 0.428 mmol), and stir at room temperature overnight. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and use V... DCM :V MeOH =10:1 extraction three times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 70-4 (280 mg, yield 55.94%).

[0807] Step 4: Synthesis of Compound 70

[0808] 70-4 (280 mg, 0.264 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. The target compound was a white solid (210 mg, yield 91.7%), [M+H]. + :862.5.

[0809] Example 71

[0810] Synthesis of (2R)-N-((2R)-6-amino-1-(3-cinnamoyl-3,8-diazabicyclo[3.2.1]octane-8-yl)-1-oxohexane-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamine hydrochloride (71)

[0811] Step 1: Synthesis of Compound 71-2

[0812] 56-1 (212 mg, 1 mmol), 32-1 (148 mg, 1 mmol), HOBT (189 mg, 1.40 mmol), and EDCI.HCl (335.9 mg, 1.75 mmol) were dissolved in 5 mL of dichloromethane. After stirring at room temperature for 5 min, triethylamine (118.2 mg, 1.168 mmol) was added, and the mixture was stirred at room temperature overnight. The reaction was quenched with saturated sodium bicarbonate aqueous solution, and the organic phase was separated. The aqueous phase was extracted three times with dichloromethane. The combined organic phases were washed with 1N dilute hydrochloric acid, concentrated, and subjected to column chromatography (V). PE :V EA =5:1), to obtain oily substance 71-2 (250mg, yield 73%).

[0813] Step 2: Synthesis of compound 71-3

[0814] Dissolve 71-2 (250 mg, 0.731 mmol) in 5 mL of ethyl acetate, add 2.5 mL of dioxane hydrochloride, stir at room temperature for 4 hours, filter, and give white solid 71-3 (195 mg, yield 96.0%).

[0815] Step 3: Synthesis of compound 71-4

[0816] Dissolve 1-1 (200 mg, 0.266 mmol) and HATU (121.22 mg, 0.319 mmol) in THF (5 mL), then add DIEA (137 mg, 1.06 mmol). Stir the reaction at room temperature for 0.5 hours, then add 71-3 (118 mg, 0.428 mmol) and stir the reaction overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and separate the aqueous phase using V... DCM :V MeOH =10:1 extraction three times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 71-4 (200 mg, yield 76.9%).

[0817] Step 4: Synthesis of Compound 71

[0818] 71-4 (200 mg, 0.205 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. The target compound 71 was a white solid (100 mg, yield 63.9%), [M+H]. + :778.4.

[0819] Example 72

[0820] Synthesis of (R)-N-((R)-6-amino-1-(4-((Z)-2-fluoro-3-phenylacrylamido)piperidin-1-yl)-1-oxohexane-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide hydrochloride (72)

[0821]

[0822] Step 1: Synthesis of Compound 72-2

[0823] Dissolve 72-1 (166 mg, 1 mmol), 7-1 (200 mg, 1 mmol), HOBT (189 mg, 1.40 mmol), and EDCI.HCl (335.9 mg, 1.75 mmol) in 5 mL of dichloromethane. Stir at room temperature for 5 min, then add triethylamine (118.2 mg, 1.168 mmol) and stir overnight at room temperature. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash with 1N dilute hydrochloric acid, concentrate, and perform column chromatography (V... PE :V EA =5:1), to obtain oily substance 72-2 (250mg, yield 72%).

[0824] Step 2: Synthesis of Compound 72-3

[0825] Dissolve 72-2 (250 mg, 0.731 mmol) in 5 mL of ethyl acetate, add 2.5 mL of dioxane hydrochloride, stir at room temperature for 4 hours, filter, and give white solid 72-3 (201 mg, yield 98.5%).

[0826] Step 3: Synthesis of compound 72-4

[0827] Dissolve 1-1 (200 mg, 0.266 mmol) and HATU (121.22 mg, 0.319 mmol) in THF (5 mL), then add DIEA (137 mg, 1.06 mmol), stir at room temperature for 0.5 hours, then add 72-3 (106 mg, 0.428 mmol), and stir at room temperature overnight. Quench the reaction with saturated sodium bicarbonate solution, separate the organic phase, and use V... DCM :V MeOH =10:1 extraction three times, combined organic phases, washed with 1N dilute hydrochloric acid, concentrated, and column chromatography (V DCM :V MeOH =20:1), yielding product 72-4 (200 mg, yield 76.6%).

[0828] Step 4: Synthesis of Compound 72

[0829] Compound 72-4 (200 mg, 0.203 mmol) was dissolved in 5 mL of ethyl acetate, and then 2.5 mL of dioxane hydrochloride was added. The mixture was stirred at room temperature for 2 hours, and then filtered to give the product. The target compound 72 was a white solid (100 mg, yield 62.5%), [M+H]. + :784.4.

[0830] Example 73

[0831] Synthesis of 1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropionylamino)-3-phenylpropionylamino)-4-methylpentanoylamino)hexanoyl)-4-((methoxycarbonyl)amino)piperidine-4-carboxylic acid (73)

[0832]

[0833] Step 1: Synthesis of compound 73-2B

[0834] 73-2A (500 mg, 1.66 mmol) and triethylamine (502 mg, 4.98 mmol) were dissolved in DCM (5 mL), and Cb2-Cl (340 mg, 2.00 mL) was added. The reaction mixture was stirred at room temperature for 3 h. After the reaction was complete as monitored by TLC, H2O (10 mL) was added to the reaction mixture, followed by extraction with DCM (20 mL). The organic phase was dried by rotary evaporation to obtain a colorless oily substance 73-2B (580 mg, yield: 80.5%), [M+H]. + :435.2.

[0835] Step 2: Synthesis of compound 73-2C

[0836] 73-2B (580 mg, 1.33 mmol) was dissolved in DCM (3 mL), and dioxane hydrochloride (3 mL, 4 M) was added to the solution. The reaction was stirred for 1 hour. The reaction was stopped after the starting material was consumed by TLC monitoring. The reaction solution was evaporated to dryness, the pH was adjusted to 9 with saturated Na2CO3, and EtOAc was added for extraction. The organic phase was dried and evaporated to dryness to obtain 73-2C as a pale yellow oil (380 mg, yield: 85.5%), [M+H]. + :335.1.

[0837] Step 3: Synthesis of compound 73-2D

[0838] 73-2C (380 mg, 0.19 mmol) was dissolved in MeCN (5 mL), and then K₂CO₃ (467 mg, 3.39 mmol) and methyl chloroformate (160 mg, 1.69 mmol) were added. The reaction mixture was stirred at room temperature for 4 h. After the reaction was completed as monitored by TLC, the reaction mixture was filtered and evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography to obtain a yellow oily substance 73-2D (350 mg, yield: 79.0%), [M+H₂]. + :393.2.

[0839] Step 4: Synthesis of Compound 73-2

[0840] 73-2D (350 mg, 0.89 mmol) was dissolved in MeOH (5 mL). Then, 50 mg of Pd(OH)₂ / C (containing 20% ​​Pd and 50% water) was added to the solution. The reaction mixture was then purged with hydrogen three times, and the reaction was stirred at room temperature for 3 hours under a hydrogen atmosphere. TLC monitoring showed that most of the starting material had been consumed, at which point the reaction was stopped. The reaction solution was directly filtered, and the filtrate was evaporated to dryness to obtain a brown oily substance 73-2 (180 mg, yield: 78.3%), [M+H]. + :259.1.

[0841] Step 5: Synthesis of Compound 73-3

[0842] Dissolve 1-1 (150 mg, 0.19 mmol) in THF (3 mL), then add HATU (91.2 mg, 0.23 mmol) and DIEA (77.4 mg, 0.60 mmol) to the solution. Stir the reaction at room temperature for 0.5 hours. Then add 73-2 (62 mg, 0.24 mmol), and continue stirring overnight. Stop the reaction when the starting material is consumed by TLC. Wash once with 1N Na₂CO₃ (10 mL) and once with 1N HCl (10 mL), respectively. Dry the organic phase and then evaporate to dryness to obtain the crude product. TLC of the crude product yields compound 73-3 as a brown solid (130 mg, yield: 65.5%), [M+H₂]. + : 994.5.

[0843] Step 6: Synthesis of Compound 73

[0844] 73-3 (130 mg, 0.15 mmol) was dissolved in DCM (1.5 mL), and TFA (0.5 mL) was added to the solution. The reaction was stirred at room temperature for 1 hour. When most of the starting material was consumed, the reaction was stopped by TLC. The reaction solution was directly concentrated, slurried with diethyl ether (10 mL), and the solid was filtered and dried to obtain the product. Target compound 73 was a grayish-white solid (55 mg, yield: 57.8%), [M+H]. + :738.4.

[0845] Example 74

[0846] Synthesis of 1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)-4-cinnamamidopiperidine-4-carboxylic acid ditrifluoroacetate (74)

[0847]

[0848] Step 1: Synthesis of Compound 74-2

[0849] 74-1 (200 mg, 0.67 mmol), HOBT (108 mg, 0.80 mmol), EDCI (180 mg, 0.94 mmol), and 32-1 (110 mg, 0.74 mmol) were dissolved in DCM (8 mL). The reaction mixture was stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] to [M+H] was increased from 100:1 to 30:1 to obtain the target compound 74-2 as a yellow oil (270 mg, yield: 94.2%). + :431.1.

[0850] Step 2: Synthesis of Compound 74-3

[0851] 74-2 (270 mg, 0.63 mmol) was dissolved in ethyl acetate (8 mL), and dioxane hydrochloride (4 M, 5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 74-3 was a white solid (221 mg, yield: 96.0%), [M+H]. +367.1. Step 3: Synthesis of compound 74-5

[0852] Dissolve 1-1 (150 mg, 0.20 mmol), HATU (91 mg, 0.24 mmol), and DIEA (77 mg, 0.60 mmol) in tetrahydrofuran (6 mL). Stir the reaction mixture at room temperature for 30 min. Then add 74-3 (80 mg, 0.22 mmol) and stir the mixture overnight at room temperature. After the reaction is complete as monitored by TLC, wash the reaction mixture once with 1N Na₂CO₃ and once with 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using eluent V. 二氯甲烷 :V 甲醇 The ratio of [M+H] was increased from 100:1 to 30:1 to obtain the target compound 74-5 as a white solid (121 mg, yield: 57.1%). + :1066.6.

[0853] Step 4: Synthesis of Compound 74

[0854] 74-5 (121 mg, 0.11 mmol) was dissolved in DCM (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. The reaction was stopped when most of the starting material was consumed, and the reaction mixture was evaporated to dryness and slurried with diethyl ether to obtain the product. Target compound 74 was a white solid (91 mg, yield: 77.2%), [M+H]. + :810.5.

[0855] Example 75

[0856] Synthesis of (R)-N-((R)-6-amino-1-(4-((S)-2-(2-fluoro-[1,1'-biphenyl]-4-yl)propamido)piperidin-1-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide dihydrochloride (75)

[0857] Step 1: Synthesis of Compound 75-2

[0858] 7-1 (0.60 g, 3.0 mmol), HOBT (0.49 g, 3.6 mmol), EDCI (0.86 g, 4.5 mmol), and 69-1 (0.80 g, 3.3 mmol) were dissolved in DCM (25 mL). The reaction mixture was stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent.二氯甲烷 :V 甲醇 The ratio of [M+H] to [M+H] was increased from 100:1 to 30:1 to obtain the target compound 75-2 as a white solid (1.21 g, yield: 94.4%). + :427.1.

[0859] Step 2: Synthesis of Compound 75-3

[0860] 75-2 (1.21 g, 2.84 mmol) was dissolved in ethyl acetate (20 mL), and dioxane hydrochloride (4 M, 10 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 75-3 was a white solid (1.01 g, yield: 98.1%), [M+H]. + :327.1. Step 3: Synthesis of compound 75-5

[0861] 1-1 (350 mg, 0.46 mmol), HATU (201 mg, 0.53 mmol), and DIEA (178 mg, 1.38 mmol) were dissolved in tetrahydrofuran (10 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 75-3 (185 mg, 0.51 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] was increased from 100:1 to 40:1 to obtain the target compound 75-5 as a white solid (281 mg, yield: 57.0%). + :1062.6.

[0862] Step 4: Synthesis of Compound 75

[0863] Compound 75-5 (281 mg, 0.26 mmol) was dissolved in ethyl acetate (8 mL), and dioxane hydrochloride (4 M, 4 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. The target compound 75 was a white solid (157 mg, yield: 63.5%), [M+H]. + :862.5.

[0864] Example 76

[0865] Synthesis of 4-amino-1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)-N-(benzofuran-5-yl)piperidine-4-carboxamide trifluoroacetate (76)

[0866] Step 1: Synthesis of Compound 76-3

[0867] 19-1 (150 mg, 0.15 mmol), HATU (68 mg, 0.18 mmol), and DIEA (39 mg, 0.30 mmol) were dissolved in tetrahydrofuran (6 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 52-1b (23 mg, 0.17 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, DCM was added to the reaction mixture, and the solution was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] to [M+H] was increased from 100:1 to 30:1 to obtain the target compound 76-3 as a white solid (118 mg, yield: 70.4%). + :1095.6.

[0868] Step 2: Synthesis of Compound 76

[0869] 76-3 (118 mg, 0.11 mmol) was dissolved in dichloromethane (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. TLC monitoring showed that most of the starting material had been consumed, at which point the reaction was stopped. The reaction mixture was evaporated to dryness, slurried with diethyl ether, and the solid was dried to obtain the product, a white solid 76 (105 mg, yield: 85.7%), [M+H]. + :795.4.

[0870] Example 77

[0871] Synthesis of (R)-N-((R)-6-amino-1-(4-((S)-2-(6-methoxynaphth-2-yl)propamido)piperidin-1-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide dihydrochloride (77)

[0872] Step 1: Synthesis of Compound 77-2

[0873] 7-1 (0.5 g, 2.5 mmol), HOBT (0.40 g, 3.0 mmol), EDCI (0.72 g, 3.75 mmol), and 70-1 (0.64 g, 2.80 mmol) were dissolved in DCM (25 mL). The reaction mixture was stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of 100:1 to 25:1 yielded a white solid 77-2 (0.82 g, yield: 82.4%), [M+H]. + :399.1.

[0874] Step 2: Synthesis of Compound 77-3

[0875] 77-2 (0.82 g, 2.06 mmol) was dissolved in ethyl acetate (20 mL), and dioxane hydrochloride (4 M, 16 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 77-3 was a white solid (0.67 g, yield: 97.2%), [M+H]. + :335.1. Step 3: Synthesis of compound 77-5

[0876] Dissolve 1-1 (250 mg, 0.33 mmol), HATU (152 mg, 0.40 mmol), and DIEA (150 mg, 1.16 mmol) in tetrahydrofuran (8 mL). Stir the reaction mixture at room temperature for 30 min. Then add 77-3 (120 mg, 0.36 mmol) and stir the mixture overnight at room temperature. After the reaction is complete as monitored by TLC, wash the reaction mixture once with 1N Na₂CO₃ and once with 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] to [M+H] was increased from 100:1 to 30:1 to obtain the target compound 77-5 as a white solid (99 mg, yield: 28.5%). + :1048.5.

[0877] Step 4: Synthesis of Compound 77

[0878] 77-5 (99 mg, 0.09 mmol) was dissolved in ethyl acetate (5 mL), and dioxane hydrochloride (4 M, 2.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 1 hour. When most of the starting material was consumed, the reaction was stopped by TLC monitoring. The reaction solution was directly filtered, and the solid was dried to obtain the product, a white solid 77 (76 mg, yield: 87.4%), [M+H]. + :848.4.

[0879] Example 78

[0880] Synthesis of 4-amino-1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)-N-(3,4-dichlorobenzyl)piperidine-4-carboxamide trifluoroacetate (78)

[0881] Step 1: Synthesis of Compound 78-3

[0882] 19-1 (150 mg, 0.15 mmol), HATU (68 mg, 0.18 mmol), and DIEA (39 mg, 0.30 mmol) were dissolved in tetrahydrofuran (6 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 78-2 (30 mg, 0.17 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, DCM was added to the reaction mixture, and the solution was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] was increased from 100:1 to 30:1 to obtain the target compound 78-3 as a white solid (109 mg, yield: 62.6%). + :1137.5.

[0883] Step 2: Synthesis of Compound 78

[0884] 78-3 (109 mg, 0.10 mmol) was dissolved in dichloromethane (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. When most of the starting material was consumed, the reaction was stopped by TLC. The reaction mixture was evaporated to dryness, slurried with diethyl ether, and the solid was dried to obtain the product. Target compound 78 was a white solid (101 mg, yield: 89.3%), [M+H]. + :837.4.

[0885] Example 79

[0886] Synthesis of 4-amino-1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)-N-(3,4-dichlorophenylethyl)piperidine-4-carboxamide trifluoroacetate (79)

[0887] Step 1: Synthesis of Compound 79-3

[0888] 19-1 (150 mg, 0.15 mmol), HATU (68 mg, 0.18 mmol), and DIEA (39 mg, 0.30 mmol) were dissolved in tetrahydrofuran (6 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 79-2 (32 mg, 0.17 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, DCM was added to the reaction mixture, and the solution was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] to [M+H] was increased from 100:1 to 30:1 to obtain the target compound 79-3 as a white solid (117 mg, yield: 66.4%). + :1151.5.

[0889] Step 2: Synthesis of Compound 79

[0890] 79-3 (117 mg, 0.10 mmol) was dissolved in dichloromethane (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. When most of the starting material was consumed, the reaction was stopped by TLC. The reaction mixture was evaporated to dryness, slurried with diethyl ether, and the solid was dried to obtain the product. Target compound 79 was a white solid (104 mg, yield: 85.7%), [M+H]. + :851.4.

[0891] Example 80

[0892] Synthesis of (2R)-N-((2R)-6-amino-1-(3-(2-(3,4-dichlorophenyl)acetamyl)-8-azabicyclo[3.2.1]oct-8-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropionylamino)-3-phenylpropionylamino)-4-methylpentanamide dihydrochloride (80)

[0893] Step 1: Synthesis of Compound 80-2

[0894] 80-1 (0.57 g, 2.5 mmol), HOBT (0.40 g, 3.0 mmol), EDCI (0.72 g, 3.75 mmol), and 59-1 (0.57 g, 2.80 mmol) were dissolved in DCM (25 mL). The reaction mixture was stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the reaction mixture was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of 100:1 to 25:1 yielded a white solid 80⁻² (0.97 g, yield: 93.3%), [M + H]. + :413.1.

[0895] Step 2: Synthesis of Compound 80-3

[0896] 80-2 (0.97 g, 2.35 mmol) was dissolved in ethyl acetate (20 mL), and dioxane hydrochloride (4 M, 16 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 h. TLC monitoring showed that most of the starting material had been consumed, at which point the reaction was stopped, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 80-3 was a white solid (0.79 g, yield: 96.1%), [M+H]. + :349.1.

[0897] Step 3: Synthesis of Compound 80-5

[0898] Dissolve 1-1 (250 mg, 0.33 mmol), HATU (152 mg, 0.40 mmol), and DIEA (150 mg, 1.16 mmol) in tetrahydrofuran (8 mL). Stir the reaction mixture at room temperature for 30 min. Then add 80-3 (126 mg, 0.36 mmol) and stir the mixture overnight at room temperature. After the reaction is complete as monitored by TLC, wash the reaction mixture once with 1N Na₂CO₃ and once with 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using eluent V. 二氯甲烷 :V 甲醇 =100:1 to 30:1 to give a white solid 80-5 (134 mg, yield: 38.6%), [M+H + :1048.5.

[0899] Step 4: Synthesis of Compound 80

[0900] Compound 80-5 (134 mg, 0.13 mmol) was dissolved in ethyl acetate (6 mL), and dioxane hydrochloride (4 M, 3 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly filtered. The solid was dried to obtain the product. Target compound 80 was a white solid (92 mg, yield: 78.2%), [M+H]. + :848.4.

[0901] Example 81

[0902] Synthesis of 4-amino-1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)-N-(2,4,6-trimethoxybenzyl)piperidine-4-carboxamide trifluoroacetate (81)

[0903] Step 1: Synthesis of compound 81-3

[0904] 19-1 (150 mg, 0.15 mmol), HATU (68 mg, 0.18 mmol), and DIEA (39 mg, 0.30 mmol) were dissolved in tetrahydrofuran (6 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 81-2 (33 mg, 0.17 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, DCM was added to the reaction mixture, and the solution was washed once with 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using V as the eluent. 二氯甲烷 :V 甲醇 The ratio of [M+H] was increased from 100:1 to 30:1 to obtain the target compound 81-3 as a white solid (123 mg, yield: 53.3%). + :1159.6.

[0905] Step 2: Synthesis of Compound 81

[0906] Compound 81-3 (123 mg, 0.11 mmol) was dissolved in dichloromethane (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. When most of the starting material was consumed, the reaction was stopped by TLC. The reaction mixture was evaporated to dryness, slurried with diethyl ether, and the solid was dried to obtain the product. Target compound 81 was a white solid (109 mg, yield: 85.5%), [M+H]. + :859.5.

[0907] Example 82

[0908] Synthesis of 2-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)-4-(aminomethyl)piperidin-4-yl)acetic acid trifluoroacetate (82)

[0909]

[0910] Step 1: Synthesis of Compound 82-2

[0911] Dissolve 8-2 (1.55 g, 8.13 mmol) in tetrahydrofuran (15 mL), then add an aqueous solution of Na2CO3 (2.15 g, 20.32 mmol) (15 mL) to the reaction mixture. Place the reaction mixture in an ice bath, then slowly add Boc2O (2.13 g, 9.76 mmol) to the reaction solution. After the addition is complete, move the reaction mixture to room temperature and stir for 6 hours. Monitor the reaction by TLC. Afterward, evaporate the organic solvent, add 15 mL of water, and extract three times with DCM. Combine the organic phases, dry them, and evaporate to dryness to obtain the crude product. Purify the crude product by column chromatography using eluent V. 二氯甲烷 :V 甲醇 The ratio of 100:1 to 30:1 yielded a white solid 82-2 (1.82 g, yield: 88.1%), [M+H]. + :255.1.

[0912] Step 2: Synthesis of compound 82-3

[0913] Dissolve 1.8 g (7.09 mmol) of 82-2 in 15 mL of ethanol, then add 15 mL of 8 M potassium hydroxide to the solution. The reaction mixture is then refluxed and stirred at 105 °C for 36 hours. Once most of the starting material has been consumed, the reaction is stopped, and the organic solvent is directly evaporated from the reaction solution to proceed to the next step, [M+H]. + :273.1.

[0914] Step 3: Synthesis of compound 82-4

[0915] After adding 15 mL of water to the crude product from the previous step, the reaction solution was placed in an ice bath. Cbz-Cl (2.18 g, 12.76 mmol) was added to the solution, and the reaction was stirred in an ice bath for 1 hour, then moved to room temperature for 5 hours. The reaction was stopped when most of the starting material was consumed by TLC monitoring. The pH was then adjusted to 1 with 1 N hydrochloric acid in an ice bath, followed by three extractions with dichloromethane / methanol = 20 / 1 (100 mL x 3). The organic phases were combined, evaporated to dryness, and purified by silica gel column chromatography. The target compound 82-4 was obtained as a pale yellow oil (1.82 g, two-step yield: 63.3%), [M+H].+ :407.1.

[0916] Step 4: Synthesis of Compound 82-5

[0917] 82-4 (1.82 g, 4.48 mmol) was dissolved in DMF (10 mL), and K2CO3 (1.21 g, 8.96 mmol) and BnBr (1.08 g, 6.27 mmol) were added sequentially to the solution. The reaction mixture was then stirred overnight at room temperature. TLC monitoring showed that most of the starting material had been consumed, at which point the reaction was stopped. The reaction mixture was poured into ice water and extracted three times with ethyl acetate (50 mL * 3). The organic phases were combined and evaporated to dryness, then passed through a silica gel column to obtain the target compound 82-5 as a pale yellow oil (2.20 g, yield: 98.9%), [M+H]. + :497.1.

[0918] Step 5: Synthesis of compound 82-6

[0919] 82-5 (500 mg, 1.0 mmol) was dissolved in ethyl acetate (5 mL), and dioxane hydrochloride (4 M, 5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 82-6 was a white solid (425 mg, yield: 97.5%), [M+H]. + 397.1. Step 6: Synthesis of compound 82-7

[0920] Dissolve 1-1 (180 mg, 0.24 mmol), HATU (110 mg, 0.29 mmol), and DIEA (77 mg, 0.60 mmol) in tetrahydrofuran (8 mL). Stir the reaction mixture at room temperature for 30 min. Then add 82-6 (112 mg, 0.26 mmol) and stir the mixture overnight at room temperature. Monitor the reaction by TLC. Afterward, wash the reaction mixture once with 25 mL of 1N Na₂CO₃ and once with 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using polar dichloromethane / methanol (V0.05). 二氯甲烷 :V 甲醇 =100:1 to 40:1) yielded a pale yellow oily substance 82-7 (125 mg, yield: 46.2%), [M+H] + :1132.4.

[0921] Step 7: Synthesis of Compound 82-8

[0922] 82-7 (125 mg, 0.11 mmol) was dissolved in methanol (12 mL), and Pd(OH)₂ / C (25 mg) was added to the reaction solution. Hydrogen gas was then introduced, and the reaction mixture was stirred at room temperature for 3 hours under hydrogen protection. The reaction was monitored by TLC, filtered, and the filtrate was evaporated to dryness and then slurried with acetonitrile to obtain a white solid 82-8 (89 mg, yield: 88.8%), [M+H]. + :908.3.

[0923] Step 8: Synthesis of Compound 82

[0924] Compound 82-8 (89 mg, 0.07 mmol) was dissolved in dichloromethane (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. When most of the starting material was consumed, the reaction was stopped, and the reaction mixture was directly evaporated to dryness and then slurried with diethyl ether to obtain the product. The target compound 82 was a white solid (73 mg, yield: 70.9%), [M+H]. + :708.2.

[0925] 1 HNMR(400MHz,CD3OD),δ7.38-7.23(m,10H),4.85(m,1H),4.72(m,1H),4.40(m,1H),4.15-3.83(m,2H),3.70-3 .52(m,3H),3.23-3.13(m,2H),3.03-2.92(m,4H),2.65(m,2H),1.71-1.46(m,15H),1.00(d,3H),0.96(d,3H).

[0926] Example 83

[0927] Synthesis of 2-(1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)-4-(aminomethyl)piperidin-4-yl)acetic acid trifluoroacetate (83)

[0928]

[0929] Step 1: Synthesis of Compound 83-2

[0930] After dissolving 83-1 (2.2 g, 2.78 mmol) in DMF (14 mL), the reaction solution was placed in an ice bath. K₂CO₃ (0.77 g, 5.56 mmol) and CH₃I (0.59 g, 4.17 mmol) were added to the solution, and the reaction was allowed to proceed at room temperature for 5 h. TLC monitoring showed that most of the starting material had been consumed. The reaction solution was then poured into water and extracted three times with dichloromethane. The organic phases were combined, washed once with saturated brine, evaporated to dryness, and purified by silica gel column chromatography. A pale yellow oil, 83-2 (1.85 g, yield: 82.6%), [M+H⁺], was obtained. + :802.2.

[0931] The solid-phase synthesis used in 83-1 was performed as follows: 5g of 2-Cl-resin was weighed and placed in a peptide synthesis tube. 25mL of dichloromethane was added, and the mixture was purged under nitrogen. Then, (Fmoc-D-Lys(Cbz)-OH (2.51g, 5mmol) and DIEA (1.29g, 10mmol) were added, and the mixture was purged under nitrogen for 1 hour. 3mL of methanol was added, and the mixture was purged under nitrogen for 30 minutes. The mixture was then dried, washed three times with DMF (25mL*3), and dried again. This solution was used directly in the next step. 25mL of piperidine / DMF (v:v = 1:4) was added to the peptide synthesis tube, and the mixture was purged under nitrogen for 30 minutes. The liquid was then dried, washed three times with DMF (25mL*3), and dried again. This solution was used directly in the next step. 50mL of... DMF, nitrogen agitation, add Fmoc-D-Leu-OH (3.53 g, 10 mmol), HATU (3.8 g, 10 mmol), DIEA (1.29 g, 10 mmol), nitrogen agitation for 1 h, drain the reaction solution, wash 3 times with DMF (25 mL * 3), drain directly for the next step; add 25 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, agitate with nitrogen for 30 min, drain the liquid, wash 3 times with DMF (25 mL * 3), drain directly for the next step; add 25 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube. DMF, purged with nitrogen, added Fmoc-D-Phe-OH (3.87 g, 10 mmol), HATU (3.8 g, 10 mmol), DIEA (1.29 g, 10 mmol), purged with nitrogen for 1 h, dried under vacuum, washed 3 times with DMF (25 mL * 3), dried under vacuum, directly used for the next step; added 50 mL piperidine / DMF (v:v = 1:4) to the peptide synthesis tube, purged with nitrogen for 30 min, dried under vacuum, washed 3 times with DMF (25 mL * 3), dried under vacuum, directly used for the next step; added 25 mL DMF to the peptide synthesis tube, purged with nitrogen, added Boc-D-Phe-OH (2 69 g (10 mmol), HATU (3.8 g, 10 mmol), DIEA (1.29 g, 10 mmol), purged under nitrogen for 1 h, dried under vacuum, washed 3 times with DMF (25 mL * 3), dried under vacuum, added MeOH (25 mL * 3), washed 3 times, dried under vacuum, added DCM (25 mL * 3), washed 3 times, dried under vacuum and used directly for the next step; added 25 mL hexafluoroisopropanol / DCM (v:v = 1:4) to the peptide synthesis tube, purged under nitrogen for 30 min, dried under vacuum, repeated 3 times, combined the resulting liquids, evaporated to dryness, to obtain 2-1 as a white solid (3.07 g, yield: 77.9%), [M+H + :788.4.

[0932] Step 2: Synthesis of compound 83-3

[0933] 83-2 (1.3 g, 1.62 mmol) was dissolved in methanol (60 mL), and Pd(OH)2 / C (0.42 g) was added to the reaction solution. Hydrogen gas was then introduced to displace the solution. The reaction mixture was stirred at room temperature under hydrogen protection for 4 hours. The reaction was monitored by TLC, followed by filtration. The filtrate was evaporated to dryness to obtain the target compound 83-3 as a light brown solid (0.91 g, yield: 84.0%), [M+H]. + :668.2.

[0934] Step 3: Synthesis of Compound 83-5

[0935] Dissolve 83-3 (0.60 g, 0.90 mmol) and 83-4 (0.13 g, 1.08 mmol) in dichloromethane (15 mL). Stir the reaction mixture at room temperature until the solvent becomes clear. Then add Na(OAc)3BH (0.38 g, 1.80 mmol) to the reaction mixture. Stir the mixture overnight at room temperature and monitor the reaction by TLC. Quench the reaction by adding a 1 N Na2CO3 aqueous solution. After removing the organic solvent by vortexing, the mixture is then subjected to V... DCM :V MeOH After three extractions at a ratio of 20:1, the organic phases were combined, dried, and then evaporated to dryness to obtain crude product 83-5, a brown oily substance [M+H]. + :770.2.

[0936] Step 4: Synthesis of compound 83-6

[0937] The crude product 83-5 (0.90 mmol) and Et3N (182 mg, 1.80 mmol) from the previous step were dissolved in DCM (20 mL). The reaction mixture was placed in an ice bath, and Boc2O (262 mg, 1.2 mmol) was added. The reaction mixture was then stirred at room temperature for 4 hours. TLC monitoring showed that most of the starting material had been consumed. The mixture was then diluted with 1 part DCM, and washed once each with 1N HCl and saturated brine. The organic phase was dried, and the filtrate was evaporated to obtain the crude product. The crude product was purified by column chromatography using polar dichloromethane / methanol eluents ranging from 80:1 to 30:1 to give a light brownish-yellow oily substance 83-6 (325 mg, two-step yield: 41.6%), [M+H]. + :770.3.

[0938] Step 5: Synthesis of compound 83-7

[0939] 83-6 (325 mg, 0.37 mmol) was dissolved in methanol (6 mL), and 4N NaOH (4 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 3 hours. The reaction was monitored by TLC. After completion, the pH was adjusted to 1 with 4N HCl in an ice bath. The methanol was then evaporated to dryness, and after adding an appropriate amount of water, the mixture was extracted three times with DCM. The organic phases were combined, dried, and evaporated to dryness to obtain a gray solid 83-7 (295 mg, yield: 92.3%), [M+H]. + :756.3.

[0940] Step 6: Synthesis of compound 83-8

[0941] 83-7 (170 mg, 0.20 mmol), HATU (91 mg, 0.24 mmol), and DIEA (77 mg, 0.60 mmol) were dissolved in tetrahydrofuran (8 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 82-6 (95 mg, 0.22 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. The reaction was monitored by TLC. Afterward, the reaction mixture was washed once with 25 mL of 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using polar dichloromethane / methanol (V... 二氯甲烷 :V 甲醇 =100:1 to 30:1) yielded a pale yellow oily substance 83-8 (100 mg, yield: 40.8%), [M+H] + :1134.4.

[0942] Step 7: Synthesis of Compounds 83-9

[0943] 83-8 (100 mg, 0.02 mmol) was dissolved in methanol (10 mL). Pd(OH)₂ / C (containing 20% ​​Pd and 50% water, 25 mg) was added to the reaction solution, followed by hydrogen purging. The reaction solution was stirred at room temperature under hydrogen protection for 3 hours. The reaction was monitored by TLC, then filtered. The filtrate was evaporated to dryness to obtain the target compound 83-9 as a light gray solid (79 mg, yield: 96.5%), [M+H]. + :910.3.

[0944] Step 8: Synthesis of Compound 83

[0945] Compound 83-9 (79 mg, 0.08 mmol) was dissolved in dichloromethane (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. When most of the starting material was consumed, the reaction was stopped, and the reaction mixture was directly evaporated to dryness and then slurried with diethyl ether to obtain the product. Target compound 83 was a white solid (61 mg, yield: 67.7%), [M+H].+ :810.3.

[0946] 1 HNMR (400MHz, CD3OD), δ7.35-7.22(m,10H),4.69(m,1H),4.38(m,1H),4.14-4.01(m,2H),3.83-3. 35(m,10H),3.26-2.76(m,11H),2.67-2.60(m,2H),1.75-1.29(m,15H),0.97(d,3H),0.93(d,3H).

[0947] Example 84

[0948] Synthesis of 2-(1-((2R,5R,8R,14R)-2-(4-aminobutyl)-8-benzyl-5-isobutyl-4,7,10-trioxo-14-phenyl-3,6,9,12-tetraazapentadecan-1-acyl)-4-(aminomethyl)piperidin-4-yl)acetic acid trifluoroacetate (84)

[0949] Step 1: Synthesis of Compound 84-1

[0950] 27-1 (180 mg, 0.23 mmol), HATU (106 mg, 0.28 mmol), and DIEA (89 mg, 0.69 mmol) were dissolved in tetrahydrofuran (8 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 82-6 (108 mg, 0.25 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. The reaction was monitored by TLC. Afterward, the reaction mixture was washed once with 25 mL of 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using polar dichloromethane / methanol (V... 二氯甲烷 :V 甲醇 =100:1 to 30:1) yielded a pale yellow oily substance to 84-1 (145 mg, yield: 54.3%), [M+H) + :1161.4.

[0951] Step 2: Synthesis of Compound 84-2

[0952] 84-1 (145 mg, 0.12 mmol) was dissolved in methanol (15 mL). Pd(OH)₂ / C (containing 20% ​​Pd and 50% water, 35 mg) was added to the reaction solution. Hydrogen gas was then introduced to displace the solution. The reaction mixture was stirred at room temperature under hydrogen protection for 3 hours. The reaction was monitored by TLC. The mixture was then filtered, and the filtrate was evaporated to dryness to obtain a white solid 84-2 (107 mg, yield: 91.5%), [M+H]. +:936.3.

[0953] Step 3: Synthesis of Compound 84

[0954] 84-2 (107 mg, 0.11 mmol) was dissolved in dichloromethane (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. When most of the starting material was consumed, the reaction was stopped, and the reaction mixture was directly evaporated to dryness and then slurried with diethyl ether to obtain the product. The target compound 84 was a white solid (92 mg, yield: 74.7%), [M+H]. + :736.2.

[0955] 1 HNMR(400MHz,CD3OD),δ7.40-7.17(m,10H),4.66(m,1H),4.58(m,1H),4.38(m,1H),4.06-3.6 3(m,4H),3.23-3.07(m,6H),3.01-2.83(m,4H),1.79-1.20(m,18H),0.98(d,3H),0.93(d,3H).

[0956] Example 85

[0957] Synthesis of (R)-N-((R)-1-(4-(acetamidomethyl)-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-6-amino-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide ditrifluoroacetate (85)

[0958]

[0959] Step 1: Synthesis of Compound 85-1

[0960] 82-4 (0.55 g, 1.35 mmol), methylamine hydrochloride (0.11 g, 1.62 mmol), HOBT (0.21 g, 1.58 mmol), and EDCI (0.41 g, 2.16 mmol) were dissolved in dichloromethane (20 mL). DIEA (0.21 g, 1.62 mmol) was then added to the solution, and the reaction mixture was stirred overnight at room temperature. TLC monitoring showed that most of the starting material had been consumed. The reaction mixture was then washed once with 25 mL of 1N Na₂CO₃ and 1N HCl, respectively. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using polar dichloromethane / methanol (V0.05). 二氯甲烷 :V 甲醇=100:1 to 30; 1) gave 85-1 (0.51 g, yield: 89.9%), a pale yellow oil, [M+H] + :420.1.

[0961] Step 2: Synthesis of Compound 85-2

[0962] 0.38 g (0.91 mmol) of 85-1 was dissolved in methanol (15 mL), and 0.13 g of Pd(OH)₂ / C was added to the reaction solution. Hydrogen gas was then introduced to displace the solution. The reaction mixture was stirred at room temperature under hydrogen protection for 3 hours. The reaction was monitored by TLC, followed by filtration. The filtrate was evaporated to dryness and then slurried with acetonitrile to obtain a brown solid, 0.24 g (yield: 92.9%), [M+H]. + :286.1.

[0963] Step 3: Synthesis of Compound 85-3

[0964] 85-2 (240 mg, 0.84 mmol) and DIEA (217 mg, 1.68 mmol) were dissolved in THF (10 mL). The reaction mixture was placed in an ice bath, and acetyl chloride (99 mg, 1.26 mmol) was added. The reaction mixture was then stirred at room temperature for 4 h. TLC monitoring showed that most of the starting material had been consumed. The solvent was then evaporated to dryness, and the mixture was reconstituted with dichloromethane. The organic phase was washed once with 25 mL of 1N Na₂CO₃ and once with 1N HCl. The organic phase was dried and evaporated to dryness. The filtrate was then evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using a solvent (V...). 二氯甲烷 :V 甲醇 =80:1 to 30:1) yielded a pale yellow oily substance 85-3 (221 mg, yield: 80.3%), [M+H] + :328.1.

[0965] Step 4: Synthesis of Compound 85-4

[0966] 85-3 (221 mg, 0.68 mmol) was dissolved in DCM (6 mL), and trifluoroacetic acid (2 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 3 h. TLC monitoring showed that most of the starting material had been consumed, at which point the reaction was stopped. The reaction mixture was directly evaporated to dryness and then slurried with diethyl ether to obtain the target compound 85-4 as a white solid (205 mg, yield: 89.0%), [M+H]. + :228.1.

[0967] Step 5: Synthesis of Compound 85-5

[0968] Dissolve 1-1 (150 mg, 0.20 mmol), HATU (91 mg, 0.24 mmol), and DIEA (77 mg, 0.60 mmol) in tetrahydrofuran (8 mL). Stir the reaction mixture at room temperature for 30 min. Then add 85-4 (75 mg, 0.22 mmol) and stir the mixture overnight at room temperature. Monitor the reaction by TLC. Afterward, wash the reaction mixture once with 25 mL of 1N Na₂CO₃ and once with 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using polar dichloromethane / methanol (V0.05). 二氯甲烷 :V 甲醇 =100:1 to 30:1) to give a white solid 85-5 (121 mg, yield: 63.2%), [M+H] + :963.3.

[0969] Step 6: Synthesis of Compound 85

[0970] 85-5 (121 mg, 0.13 mmol) was dissolved in dichloromethane (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. When most of the starting material was consumed, the reaction was stopped, and the reaction mixture was directly evaporated to dryness and then slurried with diethyl ether to obtain the product. The target compound 85 was a white solid (95 mg, yield: 76.3%), [M+H]. + :763.2.

[0971] 1 HNMR(400MHz,CD3OD),δ7.37-7.24(m,10H),4.72(m,1H),4.41(m,1H),4.12(m,1H),3.74-3.20(m,9H),3.0 3-2.92(m,4H),2.73(m,3H),2.33-2.22(m,2H),2.02(m,3H),1.83-1.41(m,13H),0.99(d,3H),0.95(d,3H).

[0972] Example 86

[0973] Synthesis of (R)-N-((R)-6-amino-1-(4-(aminomethyl)-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide trifluoroacetate (86)

[0974]

[0975] Step 1: Synthesis of Compound 86-1

[0976] Compound 85-1 (120 mg, 0.29 mmol) was dissolved in ethyl acetate (4 mL), and dioxane hydrochloride (4 M, 4 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 6 hours. The reaction was stopped when most of the starting material was consumed, and the reaction solution was directly evaporated to dryness to obtain the product. The target compound 86-1 was a pale yellow oil (95 mg, yield: 93.3%), [M+H]. + :320.1.

[0977] Step 2: Synthesis of Compound 86-2

[0978] Dissolve 1-1 (150 mg, 0.20 mmol), HATU (91 mg, 0.24 mmol), and DIEA (77 mg, 0.60 mmol) in tetrahydrofuran (8 mL). Stir the reaction mixture at room temperature for 30 min. Then add 86-1 (78 mg, 0.22 mmol) and stir the mixture overnight at room temperature. Monitor the reaction by TLC. Afterward, wash the reaction mixture once with 25 mL of 1N Na₂CO₃ and once with 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography using a polar eluent (V0.05). 二氯甲烷 :V 甲醇 =100:1 to 30:1) yielded a pale yellow oily substance 86-2 (117 mg, yield: 55.7%), [M+H] + :1055.4.

[0979] Step 3: Synthesis of Compound 86-3

[0980] 86-2 (117 mg, 0.11 mmol) was dissolved in methanol (10 mL). Pd(OH)₂ / C (containing 20% ​​Pd and 50% water, 30 mg) was added to the reaction solution. Hydrogen gas was then introduced to displace the solution. The reaction mixture was stirred at room temperature under hydrogen protection for 3 hours. The reaction was monitored by TLC. The mixture was then filtered, and the filtrate was evaporated to dryness to obtain the target compound 86-3 as a white solid (101 mg, yield: 98.9%), [M+H]. + :921.3.

[0981] Step 4: Synthesis of Compound 86

[0982] 86-3 (101 mg, 0.11 mmol) was dissolved in dichloromethane (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. When most of the starting material was consumed, the reaction was stopped, and the reaction mixture was directly evaporated to dryness and then slurried with diethyl ether to obtain the product. The target compound 86 was a white solid (95 mg, yield: 81.6%), [M+H]. + :721.2.

[0983] 1 HNMR(400MHz,CD3OD),δ7.35-7.21(m,10H),4.70(m,1H),4.38(m,1H),4.10(m,1H),3.80-3.4 1(m,4H),3.29-2.90(m,8H),2.75-2.53(m,4H),1.79-1.29(m,15H),0.98(d,3H),0.94(d,3H).

[0984] Example 87

[0985] Synthesis of (R)-N-((R)-1-(4-(acetamidomethyl)-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-6-amino-1-oxohex-2-yl)-4-methyl-2-((R)-3-phenyl-2-(2-((R)-2-phenylpropyl)amino)acetamido)pentanamide ditrifluoroacetate (87)

[0986]

[0987] Step 1: Synthesis of Compound 87-1

[0988] 27-1 (150 mg, 0.19 mmol), HATU (87 mg, 0.23 mmol), and DIEA (74 mg, 0.57 mmol) were dissolved in tetrahydrofuran (8 mL). The reaction mixture was stirred at room temperature for 30 min. Then, 85-4 (72 mg, 0.21 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. The reaction was monitored by TLC. Afterward, the reaction mixture was washed once with 25 mL of 1N Na2CO3 and once with 1N HCl. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography using polar dichloromethane / methanol (V... 二氯甲烷 :V 甲醇 =100:1 to 30:1) to give a white solid 87-1 (119 mg, yield: 62.6%), [M+H] + :991.4.

[0989] Step 2: Synthesis of Compound 87

[0990] 87-1 (119 mg, 0.12 mmol) was dissolved in dichloromethane (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. When most of the starting material was consumed, the reaction was stopped, and the reaction mixture was directly evaporated to dryness and then slurried with diethyl ether to obtain the product. The target compound 87 was a white solid (82 mg, yield: 70.9%), [M+H]. + :791.2.

[0991] 1 HNMR (400MHz, CD3OD), δ7.38-7.14(m,10H),4.69(m,1H),4.56(m,1H),4.36(m,1H),3.83-3.36(m ,7H),3.28-2.81(m,8H),2.71(m,3H),2.00(s,3H),1.75-1.31(m,18H),0.95(d,3H),0.90(d,3H).

[0992] Example 88

[0993] Synthesis of (R)-N-((R)-6-amino-1-(4-(aminomethyl)-4-(2-(deuterated methylamino)-2-oxoethyl)piperidin-1-yl)-1-oxohexane-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropylamino)-3-phenylpropylamino)-4-methylpentanamide trifluoroacetate (88)

[0994]

[0995] Step 1: Synthesis of Compound 88-1

[0996] 82-4 (0.225 g, 0.554 mmol), deuterated methylamine hydrochloride (0.055 g, 0.665 mmol), HOBT (0.105 g, 0.648 mmol), and EDCI (0.205 g, 0.887 mmol) were dissolved in dichloromethane (5 mL), and then DIEA (0.105 g, 0.665 mmol) was added. The mixture was stirred overnight at room temperature. TLC monitoring showed that most of the starting material had been consumed. The reaction solution was washed once each with 25 mL of 1N Na₂CO₃ and 1N HCl, and the organic phase was dried and concentrated to obtain the crude product. The crude product was purified by column chromatography (V... 二氯甲烷 :V 甲醇 =100:1 to 30:1), yielding a colorless oily substance 88-1 (0.18 g, yield: 77%), [M+H + :423.1.

[0997] Step 2: Synthesis of Compound 88-2

[0998] Dissolve 0.18 g (0.426 mmol) of 88-1 in 5 mL of ethyl acetate, add 2.5 mL of (4 M) dioxane hydrochloride solution, and stir at room temperature for 6 hours. Concentrate the solvent to dryness, wash with diethyl ether, and obtain 120 mg of a viscous substance, [M+H]. + :323.1.

[0999] Step 3: Synthesis of Compound 88-3

[1000] Dissolve 1-1 (150 mg, 0.199 mmol), HATU (91 mg, 0.239 mmol), and DIEA (103 mg, 0.797 mmol) in tetrahydrofuran (5 mL), stir at room temperature for 30 min, add 88-2 (85.8 mg, 0.239 mmol), stir at room temperature overnight, monitor the reaction for completeness by TLC, wash once with 25 mL of 1N Na₂CO₃ and once with 1N HCl, dry the organic phase and concentrate to obtain the crude product. Purify the crude product by column chromatography using dichloromethane / methanol (V₂CO₃ / V₂CO₃). 二氯甲烷 :V 甲醇 =100:1 to 30:1), yielding a pale yellow oily substance 88-3 (130 mg, yield: 61.6%), [M+H + :1058.

[1001] Step 4: Synthesis of Compound 88-4

[1002] Dissolve 88-3 (0.130 g, 0.123 mmol) in 5 mL of methanol, add 26 mg of Pd(OH)₂ / C (containing 20% ​​Pd and 50% water), purge with hydrogen under vacuum, and stir overnight at room temperature. Filter, concentrate, and slurry with diethyl ether to obtain the target compound 88-4 as a white powder (40 mg, yield: 35%), [M+H]. + :924.5.

[1003] Step 5: Synthesis of Compound 88

[1004] 88-4 (80 mg, 0.087 mmol) was dissolved in dichloromethane (3 mL), and then trifluoroacetic acid (1 mL) was added. The mixture was stirred at room temperature for 2 hours. When most of the starting material was consumed, the reaction was stopped, and the reaction solution was directly evaporated to dryness and then slurried with diethyl ether to obtain the product. The target compound 88 was a white solid (10 mg, yield: 22%), [M+H]. + :727.4.

[1005] 1H NMR(400MHz,CD3OD),δ7.37-7.24(m,10H),4.72(m,1H),4.41(m,1H),4.12(m,1H),3.74 -3.40(m,3H),3.23-2.92(m,6H),2.73-2.53(m,3H),1.83-1.41(m,13H),0.99(dd,6H).

[1006] Example 89

[1007] Synthesis of (R)-N-((R)-6-amino-1-(4-(aminomethyl)-4-(2-oxo-2-((2,2,2-trifluoroethyl)amino)ethyl)piperidin-1-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropionylamino)-3-phenylpropionylamino)-4-methylpentanamide trifluoroacetate (89)

[1008]

[1009] Step 1: Synthesis of Compound 89-1

[1010] 82-4 (0.20 g, 0.49 mmol), trifluoroethylamine (0.097 g, 0.98 mmol), HOBT (0.09 g, 0.59 mmol), and EDCI (0.132 g, 0.69 mmol) were dissolved in dichloromethane (6 mL). DIEA (0.21 g, 1.62 mmol) was then added to the solution, and the reaction mixture was stirred overnight at room temperature. TLC monitoring showed that most of the starting material had been consumed. The reaction mixture was then washed once with 25 mL of 1N Na₂CO₃ and 1N HCl, respectively. The organic phase was dried by rotary evaporation, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography, using a polar eluent (V...). 二氯甲烷 :V 甲醇 =100:1 to 30:1) yielded a pale yellow oily substance 89-1 (0.21 g, yield: 87.1%), [M+H] + :488.2.

[1011] Step 2: Synthesis of Compound 89-2

[1012] 89-1 (0.21 g, 0.43 mmol) was dissolved in ethyl acetate (15 mL), and HCl / EA (2.0 M, 12 mL) was added to the reaction solution. The reaction solution was stirred at room temperature under hydrogen protection for 0.5 hours, and the reaction was monitored by TLC. After rotary evaporation, a yellow oily substance 89-2 (0.18 g, yield: 96.3%) was obtained, [M+H]. + 388.2.

[1013] Step 3: Synthesis of Compound 89-3

[1014] Dissolve 1-1 (150 mg, 0.199 mmol), DIEA (103 mg, 0.8 mmol), and HATU (84.0 mg, 0.24 mmol) in THF (10 mL). Stir the reaction mixture at room temperature for 30 min, then add 89-2 (102 mg, 0.22 mmol). Stir the mixture at room temperature for 12 h. Monitor by TLC until most of the starting material is consumed. Then, evaporate the solvent to dryness, redissolve in dichloromethane, and wash the organic phase once each with 25 mL of 1N Na₂CO₃ and 1N HCl. Dry the organic phase and evaporate the filtrate to dryness to obtain the crude product. Purify the crude product by column chromatography, using a polar eluent (V0). 二氯甲烷 :V 甲醇 =80:1 to 30:1) yielded a pale yellow oily substance 89-3 (84 mg, yield: 40.3%), [M+H] + :1123.6.

[1015] Step 4: Synthesis of Compound 89-4

[1016] Dissolve 89-3 (84 mg, 0.075 mmol) in MeOH (6 mL), then add Pd(OH)₂ / C (40 mg, 10% purity) to the solution. The reaction mixture is then purged with hydrogen 3-5 times and stirred at room temperature for 16 hours. When most of the starting material has been consumed, the reaction is stopped by TLC. The reaction mixture is directly evaporated to dryness and then slurried with diethyl ether to obtain a grayish-white solid 89-4 (40 mg, yield: 52.0%), [M+H]. + :989.5.

[1017] Step 5: Synthesis of Compound 89

[1018] 89-4 (40 mg, 0.040 mmol) was dissolved in dichloromethane (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 3 hours. When most of the starting material was consumed, the reaction was stopped, and the reaction mixture was directly evaporated to dryness and then slurried with diethyl ether to obtain the product. Target compound 89 was a grayish-white solid (30 mg, yield: 70.3%), [M / 2+H]. + 395.5.

[1019] Example 90

[1020] Synthesis of (R)-N-((R)-1-(4-(N-methanesulfonyl-aminomethyl)-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-6-amino-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide ditrifluoroacetate (90)

[1021]

[1022] Step 1: Synthesis of Compound 90-1

[1023] Compound 86-3 (84 mg, 0.091 mmol), 9 mL THF, 4.5 mL DCM, 0.5 mL TEA, and a catalytic amount of DMAP were added to a reaction flask. After stirring and dissolving, 0.5 mL of methanesulfonyl chloride was added dropwise. The reaction was then maintained at 20–25 °C for 4 h. TLC monitoring showed that the starting material had reacted completely. Water was added to quench the reaction until the salt was completely dissolved. The reaction solution was then evaporated to dryness under reduced pressure at 45 °C. EA and water were added, and the mixture was stirred to dissolve and extract. The layers were separated. The EA phase was washed three times with dilute acid and water, and the layers were separated again. The EA phase was concentrated and purified by silica gel column chromatography followed by medium-pressure reversed-phase chromatography to obtain 83 mg of compound 90-1, [M+H]. + :999.3.

[1024] Step 2: Synthesis of Compound 90

[1025] Compound 90-1 (83 mg, 0.083 mmol) and 3.0 mL of DCM were added to a reaction flask and stirred until dissolved. Then, 1.0 mL of TFA was added, and the mixture was stirred at room temperature for 2 h. The mixture was then diluted with 15 mL of DCM and evaporated to dryness under reduced pressure at 40 °C. The solution was then distilled twice with 12 mL of DCM. After distillation, 8 mL of diethyl ether was added, and the mixture was stirred and beaten for 30 min. The mixture was filtered, and the filter cake was dried using an oil pump to obtain 60 mg of a white solid, compound 90, [M+H]. + :799.3.

[1026] Example 91

[1027] Synthesis of (R)-N-((R)-1-(4-guanidinyl-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-6-amino-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide trifluoroacetate (91)

[1028]

[1029] Step 1: Synthesis of Compound 91-1

[1030] 140 mg (0.152 mmol) of 86-3, 4.0 mL of THF, 4.0 mL of EtOH, and 0.5 mL of DBU were added to a reaction flask. After stirring and dissolving, 285 mg of N,N'-di-BOC-S-methylisothiourea was added. The reaction was then incubated at 30 °C for 24 h. TLC monitoring showed that the reaction proceeded completely. The solvent was evaporated under reduced pressure at 45 °C, and the solution was then purified by silica gel column chromatography followed by medium-pressure reversed-phase chromatography to obtain 130 mg of compound 91-1, [M+H]. + :1163.3.

[1031] Step 2: Synthesis of Compound 91

[1032] Compound 91-1 (130 mg, 0.112 mmol) and 4.5 mL of DCM were added to a reaction flask and stirred until dissolved. Then, 1.5 mL of TFA was added, and the mixture was stirred at room temperature for 3 h. The mixture was then diluted with 20 mL of DCM and evaporated to dryness under reduced pressure at 40 °C. The solution was then distilled twice with 15 mL of DCM. After distillation, 8 mL of diethyl ether was added, and the mixture was stirred and beaten for 30 min. The mixture was filtered, and the filter cake was dried using an oil pump to obtain 108 mg of a white solid, compound 91, [M+H]. + :763.3.

[1033] Example 92

[1034] Synthesis of (R)-N-((R)-1-(4-(N-aminocarbonyl-aminomethyl)-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-6-amino-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide ditrifluoroacetate (92)

[1035]

[1036] Step 1: Synthesis of Compound 92-1

[1037] Compound 86-3 (300 mg, 0.326 mmol), 9 mL THF, 4.5 mL DCM, and 1.5 mL LTEA were added to a reaction flask. After stirring and dissolving, 1.0 mL of phenyl chloroformate was added dropwise. The reaction was then maintained at 20–25 °C for 4 h. TLC monitoring showed that the starting material had reacted completely. Water was added to quench the reaction until the salt was completely dissolved. The reaction solution was then evaporated under reduced pressure at 45 °C to remove THF. EA (20 mL) and water (15 mL) were added, and the mixture was stirred to dissolve and extract. The layers were separated. The EA phase was washed with 1 N hydrochloric acid water (3 × 10 mL) with stirring, and the layers were separated again. The EA phase was concentrated and then subjected to silica gel column chromatography to obtain 295 mg of compound 92-1, [M+H]. +:1041.2.

[1038] Step 2: Synthesis of compound 92-2

[1039] Compound 92-1 (160 mg, 0.153 mmol) was added to a 25 mL flask, followed by 12.0 mL of isopropanol and 4.0 mL of ammonia. After stirring to dissolve, the mixture was heated to 50 °C and maintained at this temperature for 5 h. The reaction was monitored by TLC until the reactants were completely reacted. The solvent in the reaction solution was evaporated to dryness at 50 °C. Then, 15 mL of DCM was added and stirred to dissolve. The mixture was first washed with 3 mL of Na₂CO₃(aq) with stirring, resulting in layer separation. Then, it was washed with 3 mL of 1 M HCl solution with stirring, resulting in layer separation. The DCM layer was evaporated to dryness under reduced pressure at 40 °C and subjected to a reverse phase to obtain 110 mg of compound 92-2 in an oily form [M+H]. + :964.4.

[1040] Step 3: Synthesis of Compound 92

[1041] 3.0 mL of DCM was added to a reaction flask containing compound 92-2 (110 mg, 0.114 mmol), and after stirring to dissolve, 1.0 mL of TFA was added. The mixture was stirred at room temperature for 2 h, then diluted with 15 mL of DCM. The reaction solution was then evaporated to dryness under reduced pressure at 40 °C. The solution was then distilled twice with 12 mL of DCM. After distillation, 8 mL of diethyl ether was added, and the mixture was stirred and beaten for 30 min. The mixture was filtered, and the filter cake was dried using an oil pump to obtain 100 mg of a white solid compound 92, [M+H]. + :764.4.

[1042] Example 93

[1043] Synthesis of (R)-N-((R)-1-(4-(N-methoxycarbonyl-aminomethyl)-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-6-amino-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide ditrifluoroacetate (93)

[1044]

[1045] Step 1: Synthesis of Compound 93-1

[1046] Compound 86-3 (180 mg, 0.196 mmol), 94.5 mL DCM, 0.5 mL TEA, and a catalytic amount of DMAP were added to a reaction flask. After stirring and dissolving, 1.0 mL of dimethyl dicarbonate was added dropwise. The reaction was then maintained at 20–25 °C for 6 h. TLC monitoring showed that the starting material had reacted completely. Water was added to quench the reaction until the salt was completely dissolved. The reaction solution was then evaporated to dryness under reduced pressure at 45 °C. EA and water were added, and the mixture was stirred to dissolve and extract. The layers were separated. The EA phase was washed three times with dilute acid and water, and the layers were separated again. The EA phase was concentrated and purified by silica gel column chromatography followed by medium-pressure reversed-phase chromatography to obtain 120 mg of compound 93-1, [M+H]. + :979.4.

[1047] Step 2: Synthesis of Compound 93

[1048] Compound 93-1 (120 mg, 0.123 mmol) and 3.0 mL of DCM were added to a reaction flask and stirred until dissolved. Then, 1.0 mL of TFA was added, and the mixture was stirred at room temperature for 2 h. The mixture was then diluted with 15 mL of DCM and evaporated to dryness under reduced pressure at 40 °C. The solution was then distilled twice with 12 mL of DCM. After distillation, 8 mL of diethyl ether was added, and the mixture was stirred and beaten for 30 min. The mixture was filtered, and the filter cake was dried using an oil pump to obtain 95 mg of a white solid, compound 93-1, [M+H]. + :779.4.

[1049] Example 94

[1050] Synthesis of (R)-N-((R)-1-(4-(acetamidomethyl)-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-6-amino-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-3-cyclopropylpropionamide ditrifluoroacetate (94)

[1051]

[1052] Step 1: Synthesis of Compound 94-1

[1053] 5-1 (150 mg, 0.2 mmol) was dissolved in THF (3 mL), and DIEA (76 mg, 0.6 mmol) and HATU (91 mg, 0.24 mmol) were added. The mixture was heated to 30 °C and stirred for 0.5 h. 85-4 (63 mg, 0.24 mmol) was then added and stirred for 2 h. After the reaction was complete as monitored by TLC, water and dichloromethane were added for extraction. The organic phases were combined and dried to obtain the crude product. The crude product was purified by reverse-phase chromatography to obtain a light yellow, buttery compound 94-1 (30 mg, yield: 16%), [M+H]. + : 962.

[1054] Step 2: Synthesis of Compound 94

[1055] 94-1 (30 mg, 0.03 mmol) was dissolved in DCM (3 mL), and TFA (1 mL) was added. The mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC until complete. After rotary evaporation to dryness, the mixture was slurried with diethyl ether and filtered to obtain a white solid 94 (20 mg, yield: 84%), [M+H). + :761.

[1056] Example 95

[1057] Synthesis of (R)-N-((R)-1-(4-(N-ethoxycarbonyl-aminomethyl)-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-6-amino-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide ditrifluoroacetate (95)

[1058]

[1059] Step 1: Synthesis of Compound 95-1

[1060] Compound 86-3 (200 mg, 0.217 mmol), 4.5 mL THF, and 0.5 mL TEA were added to a reaction flask. After stirring and dissolving, 0.5 mL of ethyl chloroformate was added dropwise. The reaction was then incubated at 20–25 °C for 2 h. TLC monitoring showed that the starting material reacted completely. Water was added to quench the reaction until the salt was completely dissolved. The reaction solution was then evaporated to dryness under reduced pressure at 45 °C. EA and water were added, and the mixture was stirred to dissolve and extract. The layers were separated. The EA phase was washed three times with dilute acid water and separated again. The EA phase was concentrated and purified by silica gel column chromatography followed by medium-pressure reversed-phase chromatography to obtain 83 mg of compound 95-1, [M+H]. + :993.3.

[1061] Step 2: Synthesis of Compound 95

[1062] Compound 95-1 (83 mg, 0.084 mmol) and 3.0 mL of DCM were added to a reaction flask and stirred until dissolved. Then, 1.0 mL of TFA was added, and the mixture was stirred at room temperature for 2 hours. The mixture was then diluted with 15 mL of DCM and evaporated to dryness under reduced pressure at 40 °C. The solution was then distilled twice with 12 mL of DCM. After distillation, 8 mL of diethyl ether was added, and the mixture was stirred and beaten for 30 minutes. The mixture was filtered, and the filter cake was dried using an oil pump to obtain 60 mg of a white solid, compound 95-1, [M+H]. + :793.3.

[1063] Example 96

[1064] Synthesis of (R)-N-((R)-6-amino-1-(4-((3-cyclopropylurea)methyl)-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide ditrifluoroacetate (96)

[1065]

[1066] Step 1: Synthesis of Compound 96-1

[1067] 85-2 (1.0 g, 3.5 mmol) and triethylamine (763 mg, 7 mmol) were dissolved in tetrahydrofuran (20 mL). Phenyl chloroformate (655 mg, 4.2 mmol) was added to the solution under ice bath conditions. The reaction mixture was stirred overnight at room temperature. The reaction was monitored by TLC. The reaction mixture was filtered, and the filtrate was directly evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography to give a white solid 96-1 (1.1 g, yield: 78.7%), [M+H]. + 406.2.

[1068] Step 2: Synthesis of Compound 96-2

[1069] 96-1 (140 mg, 0.34 mmol) was dissolved in DMSO (3 mL), and Na2CO3 (110 mg, 1.04 mmol) and cyclopropaneamine (24 mg, 0.41 mmol) were added. The mixture was heated to 30 °C and stirred for 6 h. After the reaction was complete as monitored by TLC, saturated brine and dichloromethane were added for extraction. The organic phases were combined and dried to obtain the crude product. The crude product was purified by reverse-phase chromatography to obtain a light yellow, oily compound 96-2 (120 mg, yield: 93%), [M+H]. + 368.

[1070] Step 3: Synthesis of Compound 96-3

[1071] Dissolve 96-2 (120 mg, 0.32 mmol) in DCM (2 mL), add ethyl acetate hydrochloride (2 mL), stir at room temperature for 2 h, monitor the reaction until complete by TLC, and evaporate to dryness to obtain a light yellow buttery substance 96-3 (120 mg).

[1072] Step 4: Synthesis of Compound 96-4

[1073] Dissolve 1-1 (150 mg, 0.2 mmol) in THF (4 mL), add DIEA (68 mg, 0.6 mmol) and HATU (90 mg, 0.23 mmol), stir at room temperature for 0.5 h, add 96-3 (72 mg, 0.23 mmol), stir and react overnight. After the reaction is complete as monitored by TLC, extract with water and dichloromethane, combine the organic phases, dry to obtain crude product, and purify the crude product by scraping with a large plate (V). 二氯甲烷 :V 甲醇 =10:1), yielding a light yellow, buttery substance 96-4 (50 mg, yield: 25%), [M+H] + :1005.

[1074] Step 5: Synthesis of Compound 96

[1075] Dissolve 96-4 (50 mg, 0.05 mmol) in DCM (3 mL), add TFA (2 mL), stir at room temperature for 2 h, monitor the reaction until complete by TLC, evaporate to dryness, slurry with diethyl ether, and filter to obtain white solid 96 (38 mg, yield: 95%).

[1076] Example 97

[1077] [(R)-N-((R)-1-(4-(N-methylaminocarbonyl-aminomethyl)-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-6-amino-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide difluoroacetate](97)

[1078]

[1079] Step 1: Synthesis of Compound 97-1

[1080] Compound 86-3 (280 mg, 0.304 mmol), 6.0 mL DCM, and 0.5 mL TEA were added to a reaction flask and stirred until dissolved. Then, 80 mg of methylcarbamoyl chloride was added, and the reaction was maintained at 20–25 °C for 4 h. TLC monitoring showed that the starting material reacted completely. Water was added to quench the reaction until the salt was completely dissolved. The reaction solution was then evaporated to dryness under reduced pressure at 45 °C. EA and water were added, and the mixture was stirred to dissolve and extract. The layers were separated. The EA phase was washed three times with dilute acid and water, and the layers were separated again. The EA phase was concentrated and purified by silica gel column chromatography followed by medium-pressure reversed-phase chromatography to obtain 83 mg of compound 97-1, [M+H]. + :978.3.

[1081] Step 2: Synthesis of Compound 97

[1082] Compound 97-1 (190 mg, 0.089 mmol) and 7.5 mL of DCM were added to a reaction flask and stirred until dissolved. Then, 2.5 mL of TFA was added, and the mixture was stirred at room temperature for 2 hours. The mixture was then diluted with 20 mL of DCM and evaporated to dryness under reduced pressure at 40 °C. The solution was then distilled twice with 15 mL of DCM. After distillation, 12 mL of diethyl ether was added, and the mixture was stirred and beaten for 30 minutes. The mixture was filtered, and the filter cake was dried using an oil pump to obtain 170 mg of a white solid, compound 97-1, [M+H]. + :778.3.

[1083] Example 98

[1084] (2R)-N-[(2R)-6-amino-1-[4-(carbamoylmethyl)-4-(acetamidomethyl)piperidin-1-yl]-1-oxohexane-2-yl]-2-[(2R)-2-(2-amino-3-phenylpropylamino)-3-phenylpropylamino]-4-methylpentanamide (98)

[1085]

[1086] Step 1: Synthesis of Compound 98-4

[1087] 82-4 (500 mg, 1.2 mmol), HATU (547 mg, 1.4 mmol), DIEA (1.3 mg, 3.6 mmol), and ammonium chloride (254 mg, 4.8 mmol) were dissolved in THF (10 mL), and the mixture was stirred for 5 h. The reaction mixture was stopped after the starting materials were consumed by TLC. Water and ethyl acetate were added to the reaction solution, and the mixture was extracted, evaporated to dryness, and purified by column chromatography to obtain a light brown oily substance 98-4 (350 mg, yield: 70.0%), [M+H]. + :406.2.

[1088] Step 2: Synthesis of Compound 98-1

[1089] Dissolve 3 mL of DCM (350 mg, 0.9 mmol) in 98-4, add 1 mL of TFA, and stir for 1 h. Concentrate to obtain 250 mg of 98-1 (yield: 81.9%), [M+H]. + :306.2.

[1090] Step 3: Synthesis of Compound 98-2

[1091] Dissolve 1-1 (150 mg, 0.2 mmol) and 98-1 (73.2 mg, 0.24 mmol) in THF (3 mL), then add DIEA (77.4 mg, 0.6 mmol) and HATU (91.2 mg, 0.24 mmol) to the solution and stir for 3 h. Stop the reaction when the starting materials are consumed by TLC. Add water and ethyl acetate to the reaction solution, extract, evaporate to dryness, and purify by column chromatography to obtain a light brown oily substance 98-2 (70 mg, yield: 33.6%), [M+H]. + : 1041.6.

[1092] Step 4: Synthesis of Compound 98-3

[1093] 98-2 (70 mg, 0.06 mmol) was dissolved in MeOH (2 mL), and Pd(OH)2 / C (50 mg)Ac2O (13 mg, 0.14 mmol) and TEA (19.6 mg, 0.18 mmol) were added. The reaction mixture was stirred overnight at room temperature under a H2 (15 psi) atmosphere. After the reaction was confirmed to be complete by TLC, the mixture was filtered and evaporated to dryness to give a brown solid 98-3 (55 mg, yield: 96.1%), [M+H]. + : 949.6.

[1094] Step 5: Synthesis of Compound 98

[1095] 98-3 (55 mg, 0.05 mmol) was dissolved in DCM (1.5 mL), and then TFA (0.5 mL) was added. The reaction mixture was stirred at room temperature for 1 h. After the reaction was completed by TLC monitoring, the reaction mixture was evaporated to dryness to obtain a brown oily substance. Ether was added and the mixture was slurried to obtain a grayish-white solid 98 (30 mg, yield: 78.9%), [M+H]. + : 749.5.

[1096] Example 99

[1097] (2R)-N-[(2R)-6-amino-1-{4-[(dimethylamino)methyl]-4-[(methylcarbamoyl)methyl]piperidin-1-yl}-1-oxohexane-2-yl]-2-[(2R)-2-(2-amino-3-phenylpropylamino)-3-phenylpropylamino]-4-methylpentanylamine (99)

[1098]

[1099] Step 1: Synthesis of Compound 99-1

[1100] 86-2 (150 mg, 0.14 mmol) was dissolved in MeOH (2 mL), and Pd(OH)2 / C (50 mg) and formaldehyde aqueous solution (2 mL, 30%) were added. The reaction mixture was stirred overnight at room temperature under a H2 (15 psi) atmosphere. After the reaction was complete as monitored by TLC, the solution was filtered and evaporated to dryness to give a brown solid 99-1 (50 mg, yield: 37.8%), [M+H]. + : 949.6.

[1101] Step 2: Synthesis of Compound 99

[1102] 99-1 (50 mg, 0.05 mmol) was dissolved in DCM (1.5 mL), and then TFA (0.5 mL) was added. The reaction mixture was stirred at room temperature for 1 h. After the reaction was completed by TLC monitoring, the reaction mixture was evaporated to dryness to obtain a brown oily substance. Ether was added and the mixture was slurried to obtain a grayish-white solid 99 (20 mg, yield: 54.0%), [M+H]. + : 749.5.

[1103] Example 100

[1104] Synthesis of (2R)-N-[(2R)-6-amino-1-[4-(carbamoylmethyl)-4-(acetamidomethyl)piperidin-1-yl]-1-oxohexane-2-yl]-2-[(2R)-2-(2-amino-3-phenylpropylamino)-3-phenylpropylamino]-4-methylpentanamide (100)

[1105] Step 1: Synthesis of Compound 100-1

[1106] 83-1 (500 mg, 0.63 mmol), 85-4 (171 mg, 0.75 mmol), HATU (287 mg, 0.75 mmol), and DIEA (243 mg, 0.63 mmol) were dissolved in THF (10 mL) and the mixture was stirred for 3 h. The reaction mixture was stopped after the starting materials were consumed by TLC. Water and ethyl acetate were added to the reaction solution, and the mixture was extracted, evaporated to dryness, and purified by column chromatography to obtain a light brown oily substance 100-1 (550 mg, yield: 88.7%), [M+H]. + : 997.5.

[1107] Step 2: Synthesis of Compound 100-2

[1108] 100-1 (200 mg, 0.23 mmol) was dissolved in MeOH (5 mL), and Pd(OH)2 / C (50 mg) was added. The reaction mixture was stirred at room temperature for 6 h under a H2 (15 psi) atmosphere. After the reaction was monitored by TLC until complete, the mixture was filtered and evaporated to dryness to give a pale yellow oil, 100-2 (160 mg, yield: 80.8%), [M+H].+ : 863.5.

[1109] Step 3: Synthesis of Compound 100-4

[1110] 100-2 (110 mg, 0.12 mmol) and 100-3 (46 mg, 0.18 mmol) were dissolved in MeCN (3 mL), and TEA (39.2 mg, 0.36 mmol) was added to the solution. The reaction mixture was stirred for 2 hours. The reaction was stopped after the starting materials were consumed by TLC monitoring. The reaction solution was evaporated to dryness, and TLC yielded a pale yellow solid 100-4 (45 mg, yield: 39.4%), [M+H]. + : 951.5.

[1111] Step 4: Synthesis of Compound 100

[1112] 100-4 (45 mg, 0.05 mmol) was dissolved in DCM (1.5 mL), and then TFA (0.5 mL) was added. The reaction mixture was stirred at room temperature for 1 h. After the reaction was complete as monitored by TLC, the reaction mixture was evaporated to dryness to give a pale yellow oil. Ether was added and the mixture was slurried to give a white solid 100 (20 mg, yield: 47.6%), [M+H]. + :851.1.

[1113] Example 101

[1114] Synthesis of ((2R)-2-[(2R)-2-(2-amino-3-phenylpropylamino)-3-phenylpropylamino]-N-[(2R)-6-(dimethylamino)-1-oxo-1-{3-oxo-2,8-diazaspiro[4.5]dec-8-yl}hexane-2-yl]-4-methylpentanamide (101)

[1115] Step 1: Synthesis of Compound 101-1

[1116] After dissolving 83-1 (1 g, 1.2 mmol) and 8-2 (0.2 g, 1.5 mmol) in 20 mL of THF, DIEA (0.4 g, 1.5 mmol) and HATU (0.5 g, 4.78 mmol) were added to the solution, and the mixture was stirred for 3 h. The reaction was stopped after the starting materials were consumed by TLC monitoring. Water and ethyl acetate were added to the reaction solution, and the mixture was extracted, evaporated to dryness, and purified by column chromatography to obtain a light brown oily substance 101-1 (0.6 g, yield: 54.2%), [M+H]. + : 924.5.

[1117] Step 2: Synthesis of Compound 101-2

[1118] 101-1 (150 mg, 0.16 mmol) was dissolved in MeOH (2 mL), and Pd(OH)2 / C (50 mg) was added. The reaction mixture was stirred overnight at room temperature under a H2 (15 psi) atmosphere. After the reaction was monitored by TLC until complete, the solution was filtered and evaporated to dryness to give a brown solid 101-2 (110 mg, yield: 69.6%), [M+H]. + : 990.4.

[1119] Step 3: Synthesis of Compound 101-3

[1120] 101-2 (150 mg, 0.19 mmol) was dissolved in MeOH (2 mL), and formaldehyde aqueous solution (2 mL, 30%) and Pd(OH)2 / C (50 mg) were added. The reaction mixture was stirred overnight at room temperature under a H2 (15 psi) atmosphere. After the reaction was complete as monitored by TLC, the solution was filtered, evaporated to dryness, and purified by column chromatography to give a white solid 100-3 (60 mg, yield: 38.7%), [M+H]. + :818.5.

[1121] Step 4: Synthesis of Compound 101

[1122] Dissolve 101-3 (60 mg, 0.07 mmol) in DCM (1.5 mL), then add TFA (0.5 mL). Stir the reaction mixture at room temperature for 1 h. After the reaction is complete as monitored by TLC, evaporate the mixture to dryness to obtain a brown oily substance. Add diethyl ether and slurry to give a white solid 101 (40 mg, yield: 80.0%), [M+H]. + : 718.5.

[1123] Example 102

[1124] Synthesis of (R)-N-((R)-6-amino-1-(4-(2-amino-2-oxoethyl)-4-cyanopiperidin-1-yl)-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide ditrifluoroacetate (102)

[1125] Step 1: Synthesis of Compound 102-2

[1126] 102-1 (1 g, 4.76 mmol) and KI (157 mg, 0.95 mmol) were dissolved in tetrahydrofuran (15 mL), and the mixture was purged with nitrogen. The reaction solution was cooled to -78 °C, and LDA (2 N, 7.2 mL) was added dropwise while stirring at low temperature for 1 h. Then, 2-bromoacetamide (853 mg, 6.18 mmol) dissolved in THF (5 mL) was added dropwise, and the reaction was stirred for another 2 h. After the reaction was complete as monitored by TLC, it was quenched with saturated NHCl4 solution, washed with H2O and EA, and the organic phase was evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography with a polarity of V. 正己烷 :V 乙酸乙酯 The ratio was changed from 3:1 to 1:2, yielding a white solid 10²⁻² (324 mg, yield 25%), [M + H⁺]. + :268. Step 2: Synthesis of compound 102-3

[1127] 102-2 (150 mg, 0.56 mmol) was dissolved in DCM (2 mL), and then ethyl acetate hydrochloride (2 mL) was added. The mixture was stirred at room temperature for 3 h. After the reaction was completed by TLC monitoring, the mixture was evaporated to dryness to give a white solid 102-3 (100 mg, 87%).

[1128] Step 3: Synthesis of Compound 102-4

[1129] Dissolve 1-1 (150 mg, 0.2 mmol) in THF (4 mL), add DIEA (102 mg, 0.79 mmol) and HATU (91 mg, 0.23 mmol), heat to 30 °C and stir for 1 h, then add 103-3 (48 mg, 0.24 mmol) and stir overnight. After the reaction is complete as monitored by TLC, extract with water and dichloromethane, combine the organic phases and dry to obtain the crude product. Purify the crude product by scraping a large plate (V... 二氯甲烷 :V 甲醇 =15:1) to give a white solid 10²⁻⁴ (30 mg, yield 17%), [M+H + :904.

[1130] Step 4: Synthesis of Compound 102

[1131] 102-4 (30 mg, 0.03 mmol) was dissolved in DCM (1.5 mL), and trifluoroacetic acid (0.5 mL) was added. The mixture was stirred at room temperature for 3 h. After the reaction was complete as monitored by TLC, the solution was evaporated to dryness, slurried with diethyl ether, and filtered to obtain a white solid 102 (23 mg, 98% yield), [M+H]. + :704.

[1132] Example 103

[1133] Synthesis of (R)-N-((R)-1-(4-amino-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-6-amino-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide trifluoroacetate (103)

[1134]

[1135] Step 1: Synthesis of Compound 103-2

[1136] Compound 103-1 (1.4 g) was dissolved in dioxane (20 mL), then 2N sodium carbonate aqueous solution (20 mL) was added. Cb2Cl 1.2 g was added dropwise with stirring. After the reaction was complete as monitored by TLC, the pH was adjusted to 5-6 with 2N hydrochloric acid. DCM (3 × 30 mL) was then added and extracted three times with stirring. The layers were separated, and the DCM phase was washed once with water with stirring. The layers were then separated again. The DCM phase was concentrated and purified by silica gel column chromatography to obtain 2.0 g of compound 103-2, [M+H]. + :393.2.

[1137] Step 2: Synthesis of Compound 103-3

[1138] Compound 103-2 (830 mg) was dissolved in DCM (10 mL) with stirring. Then, 2 equivalents of DIEA, 1.2 equivalents of HOBT, and 1.4 equivalents of EDCI were added, and the mixture was activated at room temperature for 30 min with stirring. Then, 1.5 equivalents of methylamine hydrochloride were added, and the mixture was reacted overnight at room temperature. After the reaction was monitored by TLC until complete, water was added to quench the reaction, and the mixture was stirred and washed to separate the layers. The DCM phase was then washed twice with dilute acid (5 mL × 2) with stirring to separate the layers. The DCM phase was then washed once with 1N sodium carbonate aqueous solution (5 mL) with stirring to separate the layers. After concentrating the DCM phase, it was purified by silica gel column chromatography to obtain 800 mg of compound 103-3, [M+H]. + :406.2.

[1139] Step 3: Synthesis of compound 103-4

[1140] Compound 103-3 (800 mg) was dissolved in 6 mL of EA solution with stirring. Then, 6 mL of 4 M HCl / dioxane solution was added, and the mixture was heated to 35 °C and maintained at this temperature for 4 h. The reaction mixture was monitored by TLC until the reaction was complete. The solvent was evaporated under reduced pressure at 45 °C, and then the solution was evaporated three times with 10 mL of THF. After evaporation, the solution was drained using an oil pump to obtain 700 mg of a white solid compound 103-4, [M+H]. + :306.2.

[1141] Step 4: Synthesis of Compound 103-5

[1142] Compound 1-1 (1.488 g) and 25 mL of THF were added to a 50 mL bottle and stirred until dissolved. Then, 812 mg of HATU was added, followed by 805 mg of DIEA. After the addition was complete, the mixture was activated at room temperature for 30 min, then cooled to 0–5 °C and 700 mg of compound 4 was added. After the addition was complete, the cooling source was removed, and the mixture was allowed to slowly return to room temperature. The reaction was then carried out at room temperature for 18 h. LC-MS monitoring showed that compound 5 had reacted completely. The solvent in the reaction solution was evaporated under reduced pressure at 45 °C, and then 25 mL of DCM was added and stirred until dissolved. The mixture was then washed once with 10 mL of water, and the layers were separated. The DCM phase was then washed twice with 8 mL of 1 M NaCO3 (aq), and the layers were separated again. The DCM phase was then washed twice with 8 mL of 1 M HCl (aq), and the layers were separated again. After the DCM layer was concentrated, it was mixed with wet silica gel and passed through a column to obtain 1140 mg of a white solid compound 103-5, [M+H]. + :1041.1.

[1143] Step 5: Synthesis of Compound 103-6

[1144] 200 mg of compound 103-5 was added to a 25 mL bottle, followed by 3 mL of methanol, 2 mL of THF, and 4 equivalents of (Boc)2O. After stirring and dissolving, 50 mg of Pd(OH)2 / C (containing 10% Pd and 50% water) was added. The mixture was then purged with hydrogen under vacuum three times, followed by the introduction of hydrogen under balloon pressure. The mixture was kept in an oil bath at 40 °C with stirring for 2 hours. LCMS monitoring showed that the de-Cb2 reaction of the starting material was complete, but the Boc reaction had only partially occurred. The reaction solution was filtered, and catalytic amounts of DMAP, 0.5 mL of TEA, and 0.5 mL of... were added to the filtrate. Boc₂O was reacted at 25°C for 18 hours. TLC showed complete reaction of the intermediate amino compound. The reaction solution was evaporated under reduced pressure at 45°C to remove the solvent. Then, EA and water were added, and the mixture was stirred to dissolve and extract. The layers were separated. The EA phase was washed three times with dilute acid and water, and the layers were separated again. After concentration, the EA phase was purified by silica gel column chromatography followed by medium-pressure reversed-phase chromatography to obtain 130 mg of compound 10³⁻⁶, [M+H]. + :1007.3.

[1145] Step 2: Synthesis of Compound 103

[1146] Add 3.0 mL of DCM to a reaction flask containing 95 mg of compound 103-6, stir to dissolve, then add 1.0 mL of TFA, stir and react at room temperature for 2 h, then add 10 mL of DCM to dilute, then evaporate the reaction solution under reduced pressure at 40 °C, then distill twice with 10 mL of DCM. After distillation, add 6 mL of diethyl ether, stir and beat for 30 min, filter, and dry the filter cake with an oil pump to obtain 61 mg of off-white solid compound 103, [M+H]. + :707.3.

[1147] Example 104

[1148] Synthesis of (R)-N-((R)-1-(4-(N-acetyl-amino)-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-6-amino-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide ditrifluoroacetate (104)

[1149]

[1150] Step 1: Synthesis of Compound 104-1

[1151] 200 mg of compound 103-5 was added to a 25 mL bottle, followed by 3 mL of methanol and 2 mL of THF. After stirring to dissolve, 50 mg of Pd(OH)2 / C (containing 10% Pd and 50% water) was added. The mixture was purged with hydrogen under vacuum three times, and then hydrogen gas was introduced under balloon pressure. The mixture was kept in an oil bath at 35 °C with stirring for 4 h. The reaction was monitored by TLC until the reactants were completely reacted. The reaction solution was filtered, and the filtrate was evaporated to dryness under reduced pressure at 45 °C. The filtrate was then distilled three times with 10 mL of THF. After distillation, compound 104-1 was obtained as an oily substance [M+H]. + :907.3.

[1152] Step 2: Synthesis of compound 104-2

[1153] Add 4.5 mL THF, 1.5 mL TEA, and a catalytic amount of DMAP to the above reaction flask. After stirring and dissolving, add 0.5 mL acetyl chloride dropwise. Then, react at room temperature for 1 h. Monitor the reaction by TLC. If the starting material reacts completely, quench with water until the salt is completely dissolved. Then, evaporate the reaction solution to dryness under reduced pressure at 45 °C. Add EA and water, stir to dissolve and extract, and separate the layers. Wash the EA phase three times with dilute acid water and separate the layers again. After concentrating the EA phase, purify it by silica gel column chromatography followed by medium-pressure reversed-phase chromatography to obtain 80 mg of compound 10⁴⁻², [M+H]. + :949.5.

[1154] Step 2: Synthesis of Compound 104

[1155] Add 3.0 mL of DCM to a reaction flask containing 80 mg of compound 104-2, stir to dissolve, then add 1.0 mL of TFA, stir and react at room temperature for 2 h, then add 10 mL of DCM to dilute, then evaporate the reaction solution under reduced pressure at 40 °C, then distill twice with 10 mL of DCM. After distillation, add 6 mL of diethyl ether, stir and beat for 30 min, filter, and dry the filter cake with an oil pump to obtain 61 mg of off-white solid compound 104, [M+H]. +:749.5.

[1156] Example 105

[1157] (R)-N-((R)-1-(4-(N-ethoxycarbonyl-amino)-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-6-amino-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide difluoroacetate (105)

[1158]

[1159] Step 1: Synthesis of Compound 104-1

[1160] 270 mg of compound 103-5 was added to a 25 mL bottle, followed by 4.5 mL of methanol and 3 mL of THF. After stirring to dissolve, 70 mg of Pd(OH)2 / C (containing 10% Pd and 50% water) was added. Hydrogen gas was then introduced under balloon pressure, and the mixture was stirred and kept in an oil bath at 35 °C for 4 h. The reaction was monitored by TLC until the reactants were completely reacted. The reaction solution was filtered, and the filtrate was evaporated to dryness under reduced pressure at 45 °C. The filtrate was then distilled three times with 10 mL of THF. After distillation, compound 104-1 was obtained as an oily substance [M+H]. + :907.3.

[1161] Step 2: Synthesis of Compound 105-2

[1162] 4.5 mL of THF, 1.0 mL of TEA, and a catalytic amount of DMAP were added to the reaction flask containing compound 104-1. After stirring and dissolving, 0.5 mL of ethyl chloroformate was added dropwise. The reaction was then carried out at room temperature for 1 hour. TLC monitoring showed that the starting material had reacted completely. Water was added to quench the reaction until the salt was completely dissolved. The reaction solution was then evaporated to dryness under reduced pressure at 45 °C. EA and water were added, and the mixture was stirred to dissolve and extract. The layers were separated. The EA phase was washed three times with dilute acid water and separated again. The EA phase was concentrated and then purified by silica gel column chromatography followed by medium-pressure reversed-phase chromatography to obtain 115 mg of compound 105-2, [M+H]. + :979.4.

[1163] Step 3: Synthesis of Compound 105

[1164] Add 4.5 mL of DCM to a reaction flask containing 115 mg of compound 105-2, stir to dissolve, then add 1.5 mL of TFA, stir and react at room temperature for 2 h, then add 10 mL of DCM to dilute, then evaporate the reaction solution under reduced pressure at 40 °C, then distill twice with 10 mL of DCM. After distillation, add 6 mL of diethyl ether, stir and beat for 30 min, filter, and dry the filter cake with an oil pump to obtain 81 mg of off-white solid compound 105, [M+H]. + :779.4.

[1165] Example 106

[1166] Synthesis of (2R)-2-[(2R)-2-(2-amino-3-phenylpropylamino)-3-phenylpropylamino]-N-[(2R)-6-acetamido-1-oxo-1-{3-oxo-2,8-diazaspiro[4.5]dec-8-yl}hexane-2-yl]-4-methylpentanamide (106)

[1167]

[1168] Step 1: Synthesis of Compound 106-1

[1169] 101-2 (110 mg, 0.11 mmol) was dissolved in DCM (2 mL), and Ac2O (22 mg, 0.22 mmol) and TEA (35.9 mg, 0.33 mmol) were added. The reaction mixture was stirred overnight at room temperature under a H2 (15 psi) atmosphere. After the reaction was complete as monitored by TLC, the mixture was filtered and evaporated to dryness. TLC yielded a white solid 106-1 (65 mg, yield: 71.4%), [M+H]. + :832.4.

[1170] Step 2: Synthesis of Compound 106

[1171] 106-1 (65 mg, 0.07 mmol) was dissolved in DCM (1.5 mL), and then TFA (0.5 mL) was added. The reaction mixture was stirred at room temperature for 1 h. After the reaction was completed by TLC monitoring, the reaction mixture was evaporated to dryness to obtain a brown oily substance. Ether was added and the mixture was slurried to give a grayish-white solid 106 (35 mg, yield: 68.6%), [M+H]. + :732.4.

[1172] Example 107

[1173] Synthesis of (R)-N-((R)-1-(4-(N-phenoxycarbonyl-aminomethyl)-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-6-amino-1-oxohex-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide ditrifluoroacetate (107)

[1174]

[1175] Compound 92-1 (93 mg, 0.089 mmol) and 3.0 mL of DCM were added to a reaction flask and stirred until dissolved. Then, 1.0 mL of TFA was added, and the mixture was stirred at room temperature for 2 hours. The mixture was then diluted with 15 mL of DCM and evaporated to dryness under reduced pressure at 40 °C. The solution was then distilled twice with 12 mL of DCM. After distillation, 8 mL of diethyl ether was added, and the mixture was stirred and beaten for 30 minutes. The mixture was filtered, and the filter cake was dried using an oil pump to obtain 70 mg of a white solid, compound 107, [M+H]. + :841.2.

[1176] Example 108

[1177] (2R)-N-[(2R)-6-amino-1-[4-({2-[2-(2-methoxyethoxy)ethoxy]acetamido}methyl)-4-[(methylcarbamoyl)methyl]piperidin-1-yl]-1-oxohexane-2-yl]-2-[(2R)-2-(2-amino-3-phenylpropylamino)-3-phenylpropylamino]-4-methylpentanylamine (108)

[1178]

[1179] Step 1: Synthesis of Compound 108-1

[1180] 86-3 (110 mg, 0.12 mmol) and 2-(2-(2-methoxyethoxy)ethoxy)acetic acid (110 mg, 0.14 mmol) were dissolved in THF (3 mL). DIEA (46.4 mg, 0.36 mmol) and HATU (54.7 mg, 0.14 mmol) were then added to the solution, and the mixture was stirred for 1.5 h. The reaction was stopped after the starting materials were consumed by TLC. Water and ethyl acetate were added to the reaction solution, and the mixture was extracted and evaporated to dryness. TLC yielded a pale yellow solid 108-1 (60 mg, yield: 46.5%), [M+H]. + : 1081.6.

[1181] Step 2: Synthesis of Compound 108

[1182] 108-1 (60 mg, 0.05 mmol) was dissolved in DCM (1.5 mL), and then TFA (0.5 mL) was added. The reaction mixture was stirred at room temperature for 1 h. After the reaction was completed by TLC monitoring, the reaction mixture was evaporated to dryness to give a pale yellow oil. Ether was added and the mixture was slurried to give a white solid 108 (25 mg, yield: 56.8%), [M+H]. + :881.4.

[1183] Example 109

[1184] Synthesis of (2R)-N-[(2R)-6-amino-1-[4-(methoxymethyl)-4-[(methylcarbamoyl)methyl]piperidin-1-yl]-1-oxohexane-2-yl]-2-[(2R)-2-(2-amino-3-phenylpropylamino)-3-phenylpropylamino]-4-methylpentanamide (109)

[1185] Step 1: Synthesis of Compound 109-4

[1186] 102-1 (1 g, 4.76 mmol) and KI (157 mg, 0.95 mmol) were dissolved in tetrahydrofuran (15 mL), and the mixture was purged with nitrogen. The reaction solution was cooled to -78 °C, and LDA (2 N, 7.2 mL) was added dropwise while stirring at low temperature for 1 h. Then, 2-bromo-N-methylacetamide (933 mg, 6.18 mmol) dissolved in THF (5 mL) solution was added dropwise, and the reaction was stirred for another 2 h. After the reaction was complete as monitored by TLC, it was quenched with saturated NHCl4 solution, H2O was added, and the mixture was washed with EA. The organic phase was evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography with a polarity of V. 正己烷 :V 乙酸乙酯 The ratio was changed from 3:1 to 1:2, yielding a white solid 10⁹⁻⁴ (850 mg, yield 63.5%), [M + H⁺]. + :282.1.

[1187] Step 2: Synthesis of compound 109-5

[1188] 109-4 (800 mg, 2.84 mmol) was dissolved in THF (10 mL), and DIBAL-H (3 mL, 2 M) was added at -30 °C. The reaction mixture was stirred at room temperature for 1 h. After the reaction was completed by TLC monitoring, the solution was filtered, evaporated to dryness, and purified by column chromatography to obtain a pale yellow oily substance 109-5 (480 mg, yield: 55.6%), [M+H]. + :285.1.

[1189] Step 3: Synthesis of Compound 109-6

[1190] 109-5 (480 mg, 1.7 mmol) was dissolved in THF (10 mL), and LiAlH4 (393 mg, 5.4 mmol) was added at 0°C. The reaction mixture was stirred at room temperature for 1 h. After the reaction was complete as monitored by TLC, the solution was filtered, evaporated to dryness, and purified by column chromatography to give a pale yellow oil C (360 mg, yield: 74.0%), [M+H]. + :287.1.

[1191] Step 4: Synthesis of Compound 109-1

[1192] 109-6 (360 mg, 1.9 mmol) was dissolved in DCM (3 mL), and TFA (1 mL) was added. The reaction mixture was stirred at room temperature for 1 h. After the reaction was completed as monitored by TLC, the solution was filtered and evaporated to dryness to obtain a pale yellow oily substance 109-1 (230 mg, yield: 64.7%), [M+H]. + :187.1.

[1193] Step 5: Synthesis of compound 109-3

[1194] Dissolve 1-1 (100 mg, 0.13 mmol) and 109-1 (31 mg, 0.15 mmol) in THF (5 mL), then add 109-2 (27.7 mg, 0.13 mmol). Stir the reaction mixture at room temperature for 1 h. After the reaction is complete as monitored by TLC, filter, evaporate to dryness, and purify by column chromatography to obtain a pale yellow oil, 109-3 (30 mg, yield: 80.8%), [M+H]. + : 936.5.

[1195] Step 6: Synthesis of Compound 109

[1196] 109-3 (30 mg, 0.03 mmol) was dissolved in DCM (1.5 mL), and then TFA (0.5 mL) was added. The reaction mixture was stirred at room temperature for 1 h. After the reaction was completed by TLC monitoring, the reaction mixture was evaporated to dryness to obtain a pale yellow oil. Ether was added and the mixture was slurried to give a white solid 109 (4 mg, yield: 18.8%), [M+H]. + : 736.5.

[1197] Example 110

[1198] Synthesis of n-((1-(D-phenyllanol-D-phenyllanol-D-leucine-D-lysyl)-4-(2-(methylamino)-2-oxoethyl)piperidin-4-yl)methyl)palmitamide ditrifluoroacetate (110)

[1199]

[1200] Step 1: Synthesis of Compound 110-1

[1201] Pentadecanoic acid (47 mg, 0.19 mmol) was dissolved in THF (5 mL), and DIEA (62 mg, 0.48 mmol) and HATU (74 mg, 0.19 mmol) were added. The mixture was heated to 30 °C and stirred for 1 h. 86-3 (150 mg, 0.16 mmol) was added and the mixture was stirred overnight. After the reaction was complete as monitored by TLC, water and dichloromethane were added for extraction. The organic phases were combined and dried to obtain the crude product. The crude product was purified by TLC (Volume Spectroscopy). 二氯甲烷 :V 甲醇 =10:1) to give a white solid (110-1) (108 mg, yield: 57%), [M+H) + :1160.

[1202] Step 2: Synthesis of Compound 110

[1203] 110-1 (90 mg, 0.07 mmol) was dissolved in DCM (4 mL), and TFA (1.5 mL) was added. The mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC until complete. After rotary evaporation to dryness, the product was slurried with diethyl ether and filtered to obtain the target product (110) as a white solid (70 mg, yield: 93%), [M+H]. + : 960.

[1204] Example 111

[1205] Synthesis of benzyl((1-((R)-6-amino-2-((R)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentamido)hexanoyl)-4-(2-(methylamino)-2-oxoethyl)piperidin-4-yl)methyl)carbamate dihydrochloride (111)

[1206] 86-2 (60 mg, 0.057 mmol) was dissolved in ethyl acetate (2.0 mL), and dioxane hydrochloride solution (4 M, 2.0 mL) was added to the solution. The reaction mixture was then stirred at room temperature for 2 hours. When most of the starting material was consumed, the reaction was stopped by TLC. The reaction solution was diluted with 20 mL of dichloromethane, and the solution was directly evaporated to dryness. The resulting product was slurried with 15 mL of n-hexane to form a white solid. The solid was filtered and dried to obtain the final product. Target compound 111 was a white solid (41 mg, yield: 77.7%), [M+H]. + :855.3.

[1207] Example 112

[1208] (R)-N-((R)-6-amino-1-(4-((2-aminoacetamido)methyl)-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-1-oxohexyl-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide trifluoroacetate (112)

[1209]

[1210] Step 1: Synthesis of Compound 112-1

[1211] (tert-Butoxycarbonyl)glycine (24 mg, 0.14 mmol) was dissolved in THF (2 mL), and DIEA (45 mg, 0.36 mmol) and HATU (54 mg, 0.14 mmol) were added. The mixture was heated to 30 °C and stirred for 0.5 h. 86-3 (110 mg, 0.12 mmol) was added and the mixture was stirred overnight. After the reaction was complete as monitored by TLC, water and dichloromethane were added for extraction. The organic phases were combined and dried to obtain the crude product. The crude product was purified by TLC (Volume Spectrometry). 二氯甲烷 :V 甲醇 =10:1) to give a white solid (112-1) (30 mg, yield: 23%), [M+H) + :1077.

[1212] Step 2: Synthesis of Compound 112

[1213] 112-1 (30 mg, 0.02 mmol) was dissolved in DCM (3 mL), and TFA (2 mL) was added. The mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC until complete. After rotary evaporation to dryness, the mixture was slurried with diethyl ether and filtered to obtain a white solid 112 (20 mg, yield: 81%), [M+H]. + :879.

[1214] Example 113

[1215] Synthesis of (R)-N-((R)-6-amino-1-(4-((2-aminoacetamido)methyl)-4-(2-(methylamino)-2-oxoethyl)piperidin-1-yl)-1-oxohexyl-2-yl)-2-((R)-2-((R)-2-amino-3-phenylpropamido)-3-phenylpropamido)-4-methylpentanamide trifluoroacetate (113)

[1216]

[1217] Step 1: Synthesis of Compound 113-1

[1218] (tert-Butoxycarbonyl)-D-alanine (27 mg, 0.14 mmol) was dissolved in THF (2 mL), and DIEA (45 mg, 0.36 mmol) and HATU (54 mg, 0.14 mmol) were added. The mixture was heated to 30 °C and stirred for 0.5 h. 86-3 (110 mg, 0.12 mmol) was added and the mixture was stirred overnight. After the reaction was complete as monitored by TLC, water and dichloromethane were added for extraction. The organic phases were combined and dried to obtain the crude product. The crude product was purified by TLC (Volume Spectrometry). 二氯甲烷 :V 甲醇 =10:1) to give white solid 113-1 (30 mg, yield: 23%), [M+H) + :1093.

[1219] Step 2: Synthesis of Compound 113

[1220] 113-1 (30 mg, 0.02 mmol) was dissolved in DCM (3 mL), and TFA (2 mL) was added. The mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC until complete. After rotary evaporation to dryness, the mixture was slurried with diethyl ether and filtered to obtain a white solid 113 (20 mg, yield: 79%), [M+H). + : 922.

[1221] Biological test cases

[1222] Activation activity of ketogenic receptors

[1223] ForsKolin stimulates the release of cAMP in human opioid receptor-overexpressing OPRK1 cells (DiscoveRx), while opioid receptor agonists inhibit forsKolin-stimulated cAMP release. By detecting the inhibitory effect of the test compound on forsKolin-stimulated cAMP release, the agonistic activity of the compound on human opioid receptors can be determined. First, human opioid receptor-overexpressing cell lines were co-incubated with specific concentrations of forsKolin and different concentrations of the test compound. The cAMP levels in the stimulated OPRK1 cells were determined using a time-resolved fluorescence resonance energy transfer (TR-FRET) cAMP immunoassay (LANCEPerKinElmer). The specific method is as follows:

[1224] Detection buffer: 1×stimulation buffer, 500 μM IBMX, ddH2O. Compound preparation: The compound was dissolved in DMSO to prepare a stock solution with a final concentration of 10 mM. This stock solution was then diluted to a working concentration of 2 mM. The compound was serially diluted 4-fold using an Echo, with an initial concentration of 2 mM and 10 concentration gradients. 50 nL of each gradient was added to a 384 cell plate in duplicate, with a final concentration of 10 μM. The cell plates were then centrifuged at...

Claims

1. A peptide amide compound of general structural formula (I) or its solvate, pharmaceutically acceptable salt or cocrystal, characterized in that, It has the following general structural formula: （Ⅰ） in, R c Selected from C 1-6 alkyl; R d R e Each is independently selected from H; R f Selected from , ; R a and R b Together with the N atom it is attached to, it forms the following structure: ； R1 is selected from -NR7(C=O)R8; R2 is selected from H, -C 1-6 alkyl; R7 is selected from H and C. 1-6 Alkyl, 3- to 6-membered cycloalkyl; the C 1-6 The alkyl group may be further replaced by 1 to 3 deuteriums, 1 to 3 halogens, or 3 to 6-membered cycloalkyl groups; R8 is selected from -C 1-6 Alkyl, -OC 1-6 Alkyl, -OCH2CF3, -COOH , , , , , , , , , , , , , , , ; R9 is selected from halogens, hydrogen (H), deuterium, and carbon. 1-6 alkyl; R 10 R 11 Each is independently selected from H, deuterium, and halogens; R is selected from halogen, -C 1-6 Alkyl, -OC 1-6 Alkyl, wherein the C 1-6 Alkyl groups can be replaced by 1 to 3 deuterium or halogen groups; n is selected from 0, 1, 2, 3, and 4; m is selected from 0, 1, 2, and 3; p is selected from 1 and 2.

2. The peptide amide compound or its solvate, pharmaceutically acceptable salt or cocrystal according to claim 1, characterized in that, The compound is selected from one of the following structural formulas; 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 。 3. A pharmaceutical composition comprising a compound according to any one of claims 1 to 2, or a solvate thereof, a pharmaceutically acceptable salt or eutectic, and one or more pharmaceutically acceptable carriers.

4. The use of the compound or solvate thereof, pharmaceutically acceptable salt or cocrystal of any one of claims 1 to 2, or the pharmaceutical composition of claim 3, for the manufacture of a medicament for the treatment or prevention of diseases or conditions associated with κ opioid receptors in mammals, wherein the κ opioid receptor-associated diseases are selected from pain, inflammation, pruritus, edema, hyponatremia, hypokalemia, intestinal obstruction, cough, and glaucoma; wherein the pain is selected from neuropathic pain, trunk pain, visceral pain, skin pain, arthritis pain, kidney stone pain, uterine cramps, dysmenorrhea, endometriosis, indigestion, postoperative pain, eye pain, ear pain, fulminant cancer pain, and GI disorder-related pain.

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