6-diazo-5-oxo-n-leucine derivative as well as preparation method and application thereof

Abstract

According to the 6-diazo-5-oxo-n-leucine derivative, the preparation method and the application, DON is released through a breakable amido bond, immunoregulation of sepsis-related macrophages can be achieved, the safety is high, immune response of sepsis mice can be effectively inhibited through polarization of M2 type macrophages, and the 6-diazo-5-oxo-n-leucine derivative has a good application prospect. Finally, the inflammation treatment of sepsis is realized. The derivative is a metabolic drug formed by connecting DON and leucine-PEG, the drug can be self-assembled into nano-particles, release DON to play an anti-inflammatory role and reduce systemic toxicity at inflammation parts with high esterase and amidase, and the derivative has good chemical stability and drug development potential, is suitable for further pharmacological research, and can be used for preparing a drug for treating inflammation. And the compound has good synthesizability and pharmaceutical properties.

Description

Technical Field

[0001] This invention relates to the field of chemical pharmaceutical technology, and in particular to a 6-diazo-5-oxo-leucine derivative, its preparation method, and its application. Background Technology

[0002] Sepsis is a life-threatening syndrome caused by infection, characterized by dysregulation of the host response, leading to multiple organ dysfunction and high mortality. Despite advances in supportive care, effective pharmacological interventions remain elusive, primarily due to the complexity of the pathophysiology of sepsis and the lack of therapies targeting immune dysfunction.

[0003] Recent studies have highlighted the importance of immunometabolism in shaping macrophage phenotypes. Notably, pathways related to glutamate production were significantly enriched in the transcriptome of peripheral blood mononuclear cells from septic mice. Experiments revealed that glutamate promotes the conversion of macrophages to the pro-inflammatory M1 phenotype and leads to the production of pro-inflammatory cytokines, which is detrimental to sepsis treatment. The glutamate production inhibitor 6-diazo-5-oxo-leucine (DON) exhibits potent immunomodulatory activity by promoting the functional conversion of macrophages to the M2 phenotype. On the other hand, DON's poor stability and other issues limit its therapeutic efficacy when used alone.

[0004] Among the organs affected by sepsis, the lungs are particularly vulnerable, often being the first site of injury and playing a crucial role in patient survival. Therefore, there is a need to design a compound modified with DON to increase its stability and target more inflamed tissues.

[0005] It should be noted that this section is intended to provide background or context for the technical solutions of this disclosure as set forth in the claims. The description herein does not constitute an admission that it is prior art simply because it is included in this section. Summary of the Invention

[0006] The purpose of this invention is to provide a 6-diazo-5-oxo-leucine derivative, its preparation method, and its application, thereby overcoming, to at least to some extent, one or more problems caused by the limitations and defects of related technologies.

[0007] This invention first provides a 6-diazo-5-oxo-ortholeucine derivative, comprising: a compound having the structure shown in the following general formula, or an optical isomer, diastereomer, or pharmaceutically acceptable salt formed therefrom, and a pharmaceutically acceptable carrier, excipient, and excipient, as follows:

[0008] Wherein, R is selected from hydrogen atom, halogen, cyano, nitro, unsubstituted or substituted by at least one of the following groups: C1-6 alkyl, C3-8 cycloalkyl, C2-6 alkenyl, C2-6 alkoxy, C2-6 alkenyloxy, C2-6 alkoxy, C2-6 alkynyloxy, etc., wherein the following groups may be the same or different and are selected from halogen, hydroxyl, cyano, nitro and amino; unsubstituted or substituted by the following groups, such as those selected from C1-6 alkyl, C1-6 aminoalkyl, C1-6 hydroxyalkyl, C1-6 alkoxyalkyl, C1-6 cyanoalkyl, C2- 6-Alkenyl, C2-6-alkynyl, C1-6-alkylsulfonyl, C1-6-alkylcarbonyl, C2-6-alkenylcarbonyl or C2-6-alkynylcarbonyl; an ester or amide group that is unsubstituted or substituted with the following groups, wherein the following groups of the hydroxyl or amino group in the substituted ester or amide group are selected from C1-6 alkyl, C1-6 aminoalkyl, C1-6 hydroxyalkyl, C1-6 alkoxyalkyl, C1-6 cyanoalkyl, C2-6 alkenyl, C2-6-alkynyl, C1-6-alkylsulfonyl, C1-6-alkylcarbonyl, C2-6 alkenylcarbonyl or C2-6-alkynylcarbonyl.

[0009] In this invention, R is ethyl.

[0010] In this invention, the derivative is obtained by reacting 6-diazo-5-oxo-ortholeucine (DON) with CY122, and the reaction process is as follows: .

[0011] The present invention further provides a method for preparing a 6-diazo-5-oxo-ortholeucine derivative, comprising the following steps: S1, m-PEG7-CH2COOH and N-hydroxy-7-azabenzotriazole are dissolved in dry DCM, DMF is added to aid dissolution, and the reaction is carried out; S2, after the reaction mixture was cooled, N,N'-dicyclohexylcarbodiimide was added and stirring continued. Then L-leucine tert-butyl hydrochloride and DIPEA were added in sequence, and the reaction was carried out at room temperature to obtain a colorless oily compound CY122. S3, dissolve CY122 in anhydrous DCM, cool to 0°C, slowly add trifluoroacetic acid, then raise to room temperature and continue stirring to obtain a free acid intermediate; S4, the free acid and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride were dissolved in anhydrous DCM, cooled to 0°C, and 6-diazo-5-carbonyl-L-norleucine ethyl ester was added. The reaction was carried out at room temperature to obtain a 6-diazo-5-oxo-ortholeucine derivative.

[0012] In this invention, the molar ratio of m-PEG7-CH2COOH, N-hydroxy-7-azabenzotriazole and N,N'-dicyclohexylcarbodiimide is 2:3:3.

[0013] In this invention, the reaction molar ratio of L-leucine tert-butyl hydrochloride and DIPEA is 1:2.

[0014] In this invention, the dosage forms of the excipients include: oral tablets, capsules, injections, lyophilized powder injections, and sustained-release formulations.

[0015] The present invention also provides an application of a 6-diazo-5-oxo-leucine derivative, wherein the above-described derivative is used in the preparation of a drug for treating sepsis and colitis.

[0016] The technical solution provided by this invention may include the following beneficial effects: The present invention discloses a method for preparing and applying a 6-diazo-5-oxo-ortholeucine derivative. By releasing DON through a cleavable amide bond, it achieves immunomodulation of sepsis-associated macrophages with high safety. Furthermore, it effectively inhibits the immune response in septic mice through M2 macrophage polarization, ultimately achieving the treatment of sepsis inflammation. This derivative is a metabolic drug composed of DON linked to leucine-PEG. This drug can self-assemble into nanoparticles, releasing DON at sites of inflammation involving high esterase and amidase activity to exert anti-inflammatory effects and reduce systemic toxicity. It possesses good chemical stability and drug development potential, making it suitable for further pharmacological research, and exhibits good syntheticability and pharmaceutical properties. Attached Figure Description

[0017] The accompanying drawings, which are incorporated in and form part of this specification, illustrate embodiments consistent with this disclosure and, together with the description, serve to explain the principles of this disclosure. It is obvious that the drawings described below are merely some embodiments of this disclosure, and those skilled in the art can obtain other drawings based on these drawings without any inventive effort.

[0018] Figure 1 The general structural formula of the 6-diazo-5-oxo-ortholeucine derivative in this invention is shown. Figure 2 This illustrates CY122 in the present invention. 1 H- NMR Atlas; Figure 3 This illustrates CY122 in the present invention. 13 C- NMR Atlas; Figure 4 The HRMS spectrum of CY122 in this invention is shown; Figure 5 This illustrates CY128 in the present invention. 1 H- NMR Atlas; Figure 6 This illustrates CY128 in the present invention. 13 C- NMR Atlas; Figure 7 The HRMS spectrum of CY128 in this invention is shown; Figure 8 The image shows HE staining of the major organs of the mouse in this invention. Detailed Implementation

[0019] Exemplary embodiments will now be described more fully with reference to the accompanying drawings. However, these exemplary embodiments can be implemented in many forms and should not be construed as limited to the examples set forth herein; rather, they are provided so that this disclosure will be more comprehensive and complete, and will fully convey the concept of the exemplary embodiments to those skilled in the art. The described features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0020] Furthermore, the accompanying drawings are merely illustrative diagrams of embodiments of this disclosure and are not necessarily drawn to scale. The same reference numerals in the drawings denote the same or similar parts, and therefore repeated descriptions of them will be omitted. Some block diagrams shown in the drawings are functional entities and do not necessarily correspond to physically or logically independent entities.

[0021] Example 1 6-Diazo-5-oxo-leucine derivatives include: compounds having the structure shown in the following general formula, or optical isomers, diastereomers, and pharmaceutically acceptable salts formed therefrom, as well as pharmaceutically acceptable carriers, excipients, and excipients, the excipient dosage forms including: oral tablets, capsules, injections, lyophilized powders for injection, and sustained-release formulations, the general formula being as follows:

[0022] Wherein, R is selected from hydrogen atom, halogen, cyano, nitro, unsubstituted or substituted by at least one of the following groups: C1-6 alkyl, C3-8 cycloalkyl, C2-6 alkenyl, C2-6 alkoxy, C2-6 alkenyloxy, C2-6 alkoxy, C2-6 alkynyloxy, etc., wherein the following groups may be the same or different and are selected from halogen, hydroxyl, cyano, nitro and amino; unsubstituted or substituted by the following groups, such as those selected from C1-6 alkyl, C1-6 aminoalkyl, C1-6 hydroxyalkyl, C1-6 alkoxyalkyl, C1-6 cyanoalkyl, C2- 6-Alkenyl, C2-6-alkynyl, C1-6-alkylsulfonyl, C1-6-alkylcarbonyl, C2-6-alkenylcarbonyl or C2-6-alkynylcarbonyl; an ester or amide group that is unsubstituted or substituted with the following groups, wherein the following groups of the hydroxyl or amino group in the substituted ester or amide group are selected from C1-6 alkyl, C1-6 aminoalkyl, C1-6 hydroxyalkyl, C1-6 alkoxyalkyl, C1-6 cyanoalkyl, C2-6 alkenyl, C2-6-alkynyl, C1-6-alkylsulfonyl, C1-6-alkylcarbonyl, C2-6 alkenylcarbonyl or C2-6-alkynylcarbonyl.

[0023] Example 2 Preparation method of 6-diazo-5-oxo-ortholeucine derivatives The 6-diazo-5-oxo-leucine derivative is ethyl(28S,31S)-31-(4-diazo-3-oxobutyl)-28-isobutyl-26,29-dioxo-2,5,8,11,14,17,20,23-octaoxa-27,30-diazatetane-32-ester, referred to as CY128 in this application.

[0024] The preparation method includes the following steps: S1, dissolve m-PEG7-CH2COOH (100 mg, 0.24 mmol) and N-hydroxy-7-azabenzotriazole (50 mg, 0.36 mmol) in dry DCM (5 mL), and add 10% DMF (v / v) to aid dissolution.

[0025] S2, after cooling the reaction mixture to 0°C, N,N'-dicyclohexylcarbodiimide (74 mg, 0.36 mmol) was added and stirring was continued for 15 min. Subsequently, L-leucine tert-butyl hydrochloride (84 mg, 0.36 mmol) and DIPEA (93 mg, 0.72 mmol) were added sequentially. The reaction was carried out at room temperature for 12 h.

[0026] TLC monitoring showed complete conversion of the starting material. The reaction was quenched with saturated NH4Cl solution (10 mL), extracted with DCM (3 × 15 mL), and the organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column chromatography (DCM / MeOH gradient elution) to give a colorless oily compound leucine-PEG (hereinafter referred to as CY122) (74 mg, 0.127 mmol, yield 52.9%).

[0027] The proton NMR spectrum of CY122 is as follows: Figure 2 As shown, high-resolution mass spectrometry is as follows Figure 4 As shown: 1 H NMR (500 MHz, CDCl3) δ 6.65 (d, J = 8.1 Hz, 1H), 4.47 (td, J = 8.5,5.5 Hz, 1H), 3.71 (d, J = 5.8 Hz, 2H), 3.63 (d, J = 5.2 Hz, 26H), 3.55–3.50(m, 2H), 3.36 (s, 3H), 2.48 (t, J = 5.8 Hz, 2H), 1.70–1.54 (m, 2H), 1.51–1.45(m, 1H), 1.43 (s, 9H), 0.92 (dd, J = 6.5, 2.7 Hz, 6H). The carbon NMR spectrum of CY122 is as follows: Figure 3 As shown: 13 C NMR (126 MHz, CDCl3) δ173.14, 171.20, 82.83, 72.02, 70.65(11),70.50, 70.45, 67.33, 59.11, 51.34, 41.88, 37.00, 28.10(3), 25.02, 22.92,22.25. MS (ESI): m / z [M+H]+582.58. S3, CY122 (100 mg, 0.172 mmol) was dissolved in anhydrous DCM (5 mL), cooled to 0°C in an ice bath, and trifluoroacetic acid (196 mg, 1.72 mmol) was slowly added dropwise. The reaction system was brought to room temperature and stirred for 2 h. TLC showed that the starting material had completely disappeared. The reaction solution was concentrated under reduced pressure to obtain the free acid intermediate. S4, the above free acid and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (50 mg, 0.258 mmol) were dissolved in anhydrous DCM (5 mL), cooled to 0°C, and then 6-diazo-5-carbonyl-L-norleucine ethyl ester (35 mg, 0.172 mmol) were added. The reaction was carried out at room temperature for 16 h. After TLC monitoring showed that the reaction mixture was complete, the reaction mixture was quenched with saturated NH4Cl solution (10 mL), extracted with DCM (3 × 15 mL), the organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM / MeOH gradient elution) to give the yellow oily target product CY128 (70 mg, 0.099 mmol, yield 57.6%).

[0028] The proton NMR spectrum of CY128 is as follows: Figure 5 As shown, high-resolution mass spectrometry is as follows Figure 7 As shown: 1 H NMR (500 MHz, CDCl3) δ 7.00 (d, J = 7.8 Hz, 1H), 6.89 (s, 1H), 5.54(s, 1H), 4.45 (td, J = 8.3, 4.4 Hz, 1H), 4.35 (ddd, J = 9.1, 7.4, 4.9 Hz,1H), 4.14 (qd, J = 7.1, 2.8 Hz, 2H), 3.77 (t, J = 5.6 Hz, 2H), 3.74 – 3.59(m, 26H), 3.55 (dd, J = 5.9, 3.2 Hz, 2H), 3.37 (s, 3H), 2.67 (dt, J = 10.9,5.6 Hz, 1H), 2.58 (dt, J = 15.1, 5.5 Hz, 1H), 2.41 (s, 2H), 2.19 (ddt, J =12.6, 7.9, 4.8 Hz, 1H), 2.06 – 1.93 (m, 1H), 1.69 – 1.62 (m, 2H), 1.25 (t, J= 7.1 Hz, 4H), 0.98 – 0.87 (m, 6H). The carbon NMR spectrum of CY128 is as follows: Figure 6 As shown, 13C NMR (126 MHz, CDCl3) δ 193.82, 172.21, 171.54, 171.27, 71.65,70.26 (11), 70.09, 70.01, 66.94, 61.17, 58.72, 54.60, 53.40, 51.57, 40.67,36.50, 36.19, 24.45, 22.72, 21.75 (2), 13.88.MS (ESI): m / z [M+H]+707.84. Example 3 Application of 6-diazo-5-oxo-leucine derivatives: The derivatives described in this application are used in the preparation of drugs for treating sepsis, and their dosage forms can be oral tablets, capsules, injections, lyophilized powder injections, or sustained-release preparations.

[0029] Example 4 Application of 6-diazo-5-oxo-leucine derivatives: The application of the derivatives described in this application in the preparation of drugs for treating colitis, wherein the dosage form can be oral tablets, capsules, injections, lyophilized powder injections or sustained-release preparations.

[0030] Example 5 Evaluation of the effect of CY128 on the survival of septic mice This embodiment established a cecal ligation-puncture (CLP) model and conducted drug intervention and efficacy testing. Healthy 7-8 week old female C57BL / 6 mice weighing 21-28 g were used in the experiment. Before surgery, the abdominal hair of the C57BL / 6 mice was shaved. At the start of the surgery, the mice were anesthetized with isoflurane. An incision was made along the midline of the mouse's abdomen to expose the cecum, and a ligation was performed using 5-0 sutures at approximately 35% of the distance between the distal end and the base of the cecum. After ligation, a puncture was made in the cecum using a 21-gauge needle, and a drop of feces was expelled. Subsequently, the incision was sutured in layers. Postoperatively, the mice were injected with saline for fluid resuscitation. For the sham-operated group, only laparotomy was performed, and the cecum was removed from the abdominal cavity without ligation or puncture. One hour postoperatively, a cytokine (CY128, 0.3 mg / kg) was administered via the tail vein. The mortality rate of the mice was monitored within 72 hours, and the results are shown in Table 1.

[0031] Table 1. Effect of CY128 on the survival rate of septic mice

[0032] Example 6 Identification of the effect of CY128 on macrophage phenotypic transformation This experiment established an in vitro LPS-stimulated macrophage model and performed drug intervention and phenotypic identification. The mouse macrophage cell line RAW264.7 was used as the model cell. RAW264.7 cells were stimulated with LPS (100 ng / mL) for 4 hours, then treated with CY128 for 24 hours. Cells were then collected for flow cytometry. For RAW264.7 cell treatment, the cells were first washed with PBS, then stained with antibodies against the target protein membrane, and incubated at 4°C for 1 hour, gently mixing every 20 minutes. Next, fixation was performed, followed by cell membrane rupture using a membrane-breaking solution. Finally, antibodies against the target protein membrane were added for staining. The results are shown in Table 2.

[0033] Table 2 Effect of CY128 on macrophage phenotypic transformation (%, mean ± standard deviation)

[0034] Example 7 Effects of CY128 on macrophage phenotype in various organs and tissues of mice In this embodiment, lung, spleen, and ileum tissues from a CLP model were collected for macrophage marker antibody labeling. For tissue processing (spleen, lung, and ileum), the tissues were first minced with scissors, and then enzymatically dissociated at 37°C using collagenase IV and DNAase I for 30 minutes. After homogenizing the tissues using a 1 mL syringe plunger, the cells on a 70 μm filter were washed with PBS to obtain a single-cell suspension. The resulting cell suspension was then counted, and the results are shown in Table 3, for subsequent antibody staining. The antibody staining process was the same as the cell processing procedure.

[0035] Table 3 Effects of CY128 on macrophage phenotype in organs and tissues of septic mice (%, mean ± standard deviation)

[0036] Example 8 Identification of inflammatory cytokine levels in mouse serum using CY128 To further verify the regulatory role of CY128 on macrophages, this experiment measured the changes in the levels of related cytokines (TNF-α, IL-6, IFN-β, and IL-1β) secreted by macrophages in the serum of septic mice. The day before the experiment, Capture Antibody was used to seal the plates and incubate overnight at 4°C. The next day, the plates were washed three times with Wash Buffer and then incubated with ELISPOT Diluent at room temperature for 1 hour. After incubation, standards and samples were added, the plates were sealed, and incubated at room temperature for 2 hours. Detection Antibody was then added and incubated at room temperature for 1 hour. Streptavidin-HRP was then added and incubated at room temperature for 30 minutes, followed by TMB incubation for 15 minutes. Finally, Stop Solution was added, and absorbance was measured at 450 nm. The results are shown in Table 4.

[0037] Table 4. Effects of CY128 on serum inflammatory cytokine levels in septic mice. (pg / mL, mean ± standard deviation)

[0038] Example 9 Identification of inflammatory cytokine levels in mouse bronchoalveolar lavage fluid (BALF) using CY128 In this experimental case, the BALF of the constructed CLP model was tested using ELISA. Twenty-four hours after CLP modeling, C57BL / 6 mice were euthanized, their limbs were immobilized, and the neck surface was disinfected with alcohol. The muscle and adipose tissue around the trachea were carefully separated using scissors and forceps to expose the trachea. A small triangular incision was then made above the trachea. A gavage needle was inserted into the trachea and secured with sutures to prevent dislodgement. 500 μl of PBS was slowly injected into the trachea using a 1 ml syringe while gently massaging the lungs. The fluid was aspirated after 20 seconds. This flushing process was usually repeated three times. The results are shown in Table 5.

[0039] Table 5. Effects of CY128 on the levels of inflammatory cytokines in BALF of septic mice (pg / mL, mean ± standard deviation)

[0040] Example 10 Verification of the regulatory effect of CY128 on liver and kidney function indicators in septic mice This embodiment established a mouse model of sepsis. Serum was collected to detect liver function (ALP, ALT, AST) and kidney function (CRE, UA, UREA) levels. The mouse's head was fixed by holding the skin between its ears with the left hand. To prevent clotting, the whiskers were trimmed. Gently pressure was applied to both sides of the neck to cause the eyeballs to become congested and protrude. The eyeballs were quickly removed using curved forceps, and the eye sockets were immediately tilted downwards to allow blood to drip into EP tubes. The serum was obtained by incubating at 37°C for 1 hour and centrifuging at 3000 rpm for 20 minutes. Furthermore, HE staining of the major organs (heart, liver, spleen, and kidneys) showed no obvious inflammatory infiltration, necrosis, or structural damage, indicating that CY128 has good systemic tolerability and biosafety, as shown in Table 6. Figure 7 As shown.

[0041] Table 6 Effects of CY128 on liver and kidney function (mean ± standard deviation)

[0042] Compared with related technologies, the 6-diazo-5-oxo-ortholeucine derivative provided by this invention is an immunomodulatory drug linked to DON and CY122, which has the following beneficial effects: (1) Low toxicity and high stability: It has high esterase and amidase in the inflammatory environment. Immunomodulatory drugs that link DON to CY122 can release more DON under the action of these enzymes in the inflammatory conditions, which can achieve selective delivery to septic lung tissue, release a large amount of DON in the lung tissue, significantly improve the selectivity of treatment, and reduce the toxicity to normal tissues. (2) Significant anti-inflammatory activity: It significantly prolongs the survival time of mice in the CLP model. It inhibits the innate immune system, enhances the anti-inflammatory immune response, and significantly alleviates the progression of sepsis by inducing macrophages to the anti-inflammatory M2 type polarization; (3) The mechanism of immune metabolic reprogramming is clear: the survival of septic mice is enhanced by inhibiting the level of glutamate in the systemic circulation and stimulating the transformation of macrophage anti-inflammatory phenotype, which provides a theoretical basis for a deeper understanding of immunotherapy for sepsis; (4) Reduce toxic exposure and increase safety window: It has higher selectivity for septic lung tissue, significantly reduces toxic exposure to other organs without reducing efficacy, has better safety, and has a wider clinical application potential; (5) Clear chemical structure and good development prospects: The synthesis route is clear and the structure is stable. It has excellent chemical modification potential and pharmacokinetic characteristics, making it suitable for further pharmacological evaluation and industrial development, providing a new strategy for targeted treatment of inflammatory models such as sepsis.

[0043] The embodiments of the present invention have been described above with reference to the accompanying drawings. However, the present invention is not limited to the specific embodiments described above. The specific embodiments described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of the present invention without departing from the spirit and scope of the claims. Those skilled in the art can combine and integrate the different embodiments or examples described in this specification, and these all fall within the protection scope of the present invention.

Claims

1,6-diazo-5-oxo-ortholeucine derivative, characterized in that, The derivatives include: compounds having the structure shown in the following general formula, or optical isomers, diastereomers, and pharmaceutically acceptable salts formed therefrom, as well as pharmaceutically acceptable carriers, excipients, and excipients, as follows: Wherein, R is selected from hydrogen atom, halogen, cyano, nitro, unsubstituted or substituted by at least one of the following groups: C1-6 alkyl, C3-8 cycloalkyl, C2-6 alkenyl, C2-6 alkoxy, C2-6 alkenyloxy, C2-6 alkoxy, C2-6 alkynyloxy, etc., wherein the following groups may be the same or different and are selected from halogen, hydroxyl, cyano, nitro and amino; unsubstituted or substituted by the following groups, such as those selected from C1-6 alkyl, C1-6 aminoalkyl, C1-6 hydroxyalkyl, C1-6 alkoxyalkyl, C1-6 cyanoalkyl, C2- 6-Alkenyl, C2-6-alkynyl, C1-6-alkylsulfonyl, C1-6-alkylcarbonyl, C2-6-alkenylcarbonyl or C2-6-alkynylcarbonyl; an ester or amide group that is unsubstituted or substituted with the following groups, wherein the following groups of the hydroxyl or amino group in the substituted ester or amide group are selected from C1-6 alkyl, C1-6 aminoalkyl, C1-6 hydroxyalkyl, C1-6 alkoxyalkyl, C1-6 cyanoalkyl, C2-6 alkenyl, C2-6-alkynyl, C1-6-alkylsulfonyl, C1-6-alkylcarbonyl, C2-6 alkenylcarbonyl or C2-6-alkynylcarbonyl.

2. The 6-diazo-5-oxo-ortholeucine derivative according to claim 1, characterized in that, R stands for ethyl.

3. The method for preparing the 6-diazo-5-oxo-ortholeucine derivative as described in claim 1 or 2, characterized in that, The derivative was obtained by reacting 6-diazo-5-oxo-ortholeucine (DON) with CY122, and the reaction process is as follows: 。 4. The method for preparing the 6-diazo-5-oxo-ortholeucine derivative according to claim 3, characterized in that, Includes the following steps: S1, m-PEG7-CH2COOH and N-hydroxy-7-azabenzotriazole are dissolved in dry DCM, DMF is added to aid dissolution, and the reaction is carried out; S2, after the reaction mixture was cooled, N,N'-dicyclohexylcarbodiimide was added and stirring continued. Then L-leucine tert-butyl hydrochloride and DIPEA were added in sequence, and the reaction was carried out at room temperature to obtain a colorless oily compound CY122. S3, dissolve CY122 in anhydrous DCM, cool to 0°C, slowly add trifluoroacetic acid, then raise to room temperature and continue stirring to obtain a free acid intermediate; S4, the free acid and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride were dissolved in anhydrous DCM, cooled to 0°C, and 6-diazo-5-carbonyl-L-norleucine ethyl ester was added. The reaction was carried out at room temperature to obtain a 6-diazo-5-oxo-ortholeucine derivative.

5. The method for preparing the 6-diazo-5-oxo-ortholeucine derivative according to claim 4, characterized in that, The molar ratio of m-PEG7-CH2COOH, N-hydroxy-7-azabenzotriazole and N,N'-dicyclohexylcarbodiimide is 2:3:

3.

6. The method for preparing the 6-diazo-5-oxo-ortholeucine derivative according to claim 5, characterized in that, The reaction molar ratio of L-leucine tert-butyl hydrochloride and DIPEA is 1:

2.

7. The 6-diazo-5-oxo-ortholeucine derivative according to claim 1, characterized in that, Excipient dosage forms include: oral tablets, capsules, injections, lyophilized powder for injection, and sustained-release formulations.

8. The application of 6-diazo-5-oxo-ortholeucine derivatives, characterized in that, Use of the derivative of claim 1 or 2 in the preparation of a medicament for treating sepsis and colitis.

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