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CN-121991064-A - Covalent inhibitor derivative of bio-orthogonal group, preparation method and application thereof

CN121991064ACN 121991064 ACN121991064 ACN 121991064ACN-121991064-A

Abstract

The invention belongs to the technical field of chemical synthesis, and particularly discloses a covalent inhibitor derivative of a bio-orthogonal group, a preparation method and application thereof, wherein the covalent inhibitor derivative of the bio-orthogonal group takes a KRASG12C inhibitor sotorasib as a reconstruction molecule, polyethylene glycol with different chain lengths is introduced as fusion protein, and different bio-orthogonal groups are connected to obtain a sotorasib derivative, the bio-orthogonal group comprises dibenzo ring Xin Guian, alkynyl and tetrazinyl, and the chain length of the polyethylene glycol is 4-16. The invention adopts the covalent inhibitor derivative of the bio-orthogonal group, the preparation method and the application thereof, and provides a new way for solving the problems of target selection limitation and heterogeneity in tumor immunotherapy by designing and synthesizing the covalent inhibitor modified by the bio-orthogonal group to induce and generate tumor specific new antigen.

Inventors

  • DING WEN
  • Zhu Sanyong
  • LI ZHENZHEN
  • DANG YONGJUN

Assignees

  • 重庆医科大学

Dates

Publication Date
20260508
Application Date
20260123

Claims (10)

  1. 1. The covalent inhibitor derivative of bio-orthogonal group is characterized in that KRASG12C inhibitor sotorasib is taken as an remodel molecule, polyethylene glycol with different chain lengths is introduced as fusion protein, and different bio-orthogonal groups are connected to obtain sotorasib derivative; the bioorthogonal groups include dibenzo ring Xin Guian, alkynyl and tetrazinyl; the chain length of the polyethylene glycol is 4-16.
  2. 2. The covalent inhibitor derivative of bio-orthogonal groups of claim 1, wherein said sotorasib derivative comprises sotorasib-PEG 4-alkynyl, sotorasib-PEG 5-alkynyl, sotorasib-PEG 4-tetrazine, sotorasib-PEG4-DBCO, sotorasib-PEG10-DBCO, sotorasib-PEG16-DBCO; the sotorasib-PEG 4-alkynyl has the structural formula: ; the sotorasib-PEG 5-alkynyl has the structural formula: ; The sotorasib-PEG 4-tetrazine has the structural formula: ; The sotorasib-PEG4-DBCO has the structural formula: ; The structural formulas of sotorasib-PEG10-DBCO and sotorasib-PEG16-DBCO are as follows: ; wherein n is 10 or 16.
  3. 3. A method for preparing a covalent inhibitor derivative of a bio-orthogonal group, comprising the steps of: S1, dissolving one or more of a KRASG12C inhibitor sotorasib, bromoacetic acid tert-butyl ester, potassium carbonate or 2-bromoethyl (methyl) carbamic acid tert-butyl ester in N, N-dimethylformamide according to a proportion, and reacting to obtain an intermediate; s2, sequentially adding dichloromethane and trifluoroacetic acid, and reacting to obtain an intermediate; s3, adding the intermediate obtained from the compounds a and S2 into an N, N-dimethylformamide solvent or a dichloromethane solvent, and reacting to obtain a target product; The compound a is a small molecular compound of PEG-connected bio-orthogonal group or amino-polyethylene glycol-tert-butyl propionate.
  4. 4. A method for the preparation of a covalent inhibitor derivative of a bioorthogonal group according to claim 3, comprising in particular the steps of: S1, dissolving a KRASG12C inhibitor sotorasib, bromobutyl acetate and potassium carbonate in N, N-dimethylformamide according to the dosage ratio of 0.09mmol to 0.18mmol to 1mL, and reacting for 2h at the temperature of 55 ℃ to obtain an intermediate 2; s2, dissolving the intermediate 2 in dichloromethane, adding trifluoroacetic acid, and reacting for 3 hours at room temperature, wherein the volume ratio of the dichloromethane to the trifluoroacetic acid is 3:1, so as to obtain an intermediate 3; s3, adding the compound a, the intermediate 3, the HATU and the DIPEA into N, N-dimethylformamide according to the dosage ratio of 0.07mmol to 0.14mmol to 50 mu L to 1mL, and stirring and reacting for 1h at room temperature to obtain a target product sotorasib-PEG4-DBCO; Wherein, the compound a is polyethylene glycol with the chain length of 4 and is connected with dibenzo ring Xin Guian.
  5. 5. A method for the preparation of a covalent inhibitor derivative of a bioorthogonal group according to claim 3, comprising in particular the steps of: S1, dissolving a KRASG12C inhibitor sotorasib, bromobutyl acetate and potassium carbonate in N, N-dimethylformamide according to the dosage ratio of 0.09mmol to 0.18mmol to 1mL, and reacting for 2h at the temperature of 55 ℃ to obtain an intermediate 2; S2, dissolving the intermediate 2 in dichloromethane, adding trifluoroacetic acid, and reacting for 3 hours at room temperature, wherein the volume ratio of the dichloromethane to the trifluoroacetic acid is 3:1, so as to obtain an intermediate 3; S3, adding a compound a, an intermediate 3, HATU and DIPEA into N, N-dimethylformamide according to the dosage ratio of 0.08mmol to 0.13mmol to 30 mu L to 1mL, stirring and reacting overnight at room temperature to obtain an intermediate 4, then adding trifluoroacetic acid and dichloromethane, reacting for 3 hours at room temperature to obtain an intermediate 5, then adding a compound b, HATU, DIPEA according to the dosage ratio of 0.05mmol to 0.06mmol to 14 mu L, and stirring and reacting overnight at room temperature to obtain a target product; Wherein, the compound a is amino-polyethylene glycol-tert-butyl propionate with the chain length of 10 or 16, and the compound b is aza dibenzo ring Xin Guian.
  6. 6. A method of preparing a covalent inhibitor derivative of a bioorthogonal group according to claim 3, comprising the steps of: S1, dissolving a KRASG12C inhibitor sotorasib, bromobutyl acetate and potassium carbonate in N, N-dimethylformamide according to the dosage ratio of 0.09mmol to 0.18mmol to 1mL, and reacting for 2h at the temperature of 55 ℃ to obtain an intermediate 2; S2, dissolving the intermediate 2 in dichloromethane, adding trifluoroacetic acid, and reacting for 3 hours at room temperature, wherein the volume ratio of the dichloromethane to the trifluoroacetic acid is 3:1, so as to obtain an intermediate 3; S3, adding the compound a, the intermediate 3, the HATU and the DIPEA into N, N-dimethylformamide according to the dosage ratio of 0.1mmol to 0.08mmol to 0.16mmol to 55 mu L to 1mL, and reacting for 4 hours at room temperature to obtain a target product sotorasib-PEG 4-alkynyl; Wherein, the compound a is polyethylene glycol connecting alkynyl with chain length of 4.
  7. 7. A method of preparing a covalent inhibitor derivative of a bioorthogonal group according to claim 3, comprising the steps of: S1, dissolving a KRASG12C inhibitor sotorasib, bromobutyl acetate and potassium carbonate in N, N-dimethylformamide according to the dosage ratio of 0.09mmol to 0.18mmol to 1mL, and reacting for 2h at the temperature of 55 ℃ to obtain an intermediate 2; S2, dissolving the intermediate 2 in dichloromethane, adding trifluoroacetic acid, and reacting for 3 hours at room temperature, wherein the volume ratio of the dichloromethane to the trifluoroacetic acid is 3:1, so as to obtain an intermediate 3; s3, adding the compound a, the intermediate 3, the HATU and the DIPEA into N, N-dimethylformamide according to the dosage ratio of 0.08mmol to 0.07mmol to 0.14mmol to 48 mu L to 1mL, and reacting for 4 hours at room temperature to obtain a target product sotorasib-PEG 5-alkynyl; wherein, the compound a is polyethylene glycol connecting alkynyl with chain length of 5.
  8. 8. A method of preparing a covalent inhibitor derivative of a bioorthogonal group according to claim 3, comprising the steps of: S1, dissolving a KRASG12C inhibitor sotorasib, tert-butyl 2-bromoethyl (methyl) carbamate and potassium carbonate in N, N-dimethylformamide according to the dosage ratio of 0.09mmol to 0.18mmol to 1mL, and reacting for 2 hours at the temperature of 55 ℃ to obtain an intermediate 8; S2, dissolving the intermediate 8 in dichloromethane, adding trifluoroacetic acid, and reacting for 3 hours at room temperature, wherein the volume ratio of the dichloromethane to the trifluoroacetic acid is 3:1, so as to obtain an intermediate 9; S3, adding the compound a and the intermediate 9 into dichloromethane according to the dosage ratio of 0.05mmol to 1mL, and reacting for 5h at room temperature to obtain a target product sotorasib-PEG 4-tetrazine; Wherein, the compound a is polyethylene glycol with the chain length of 4 and is connected with tetrazinyl.
  9. 9. Use of a target product prepared by a preparation method of the covalent inhibitor derivative of a bioorthogonal group according to any one of claims 1-2 or the covalent inhibitor derivative of a bioorthogonal group according to any one of claims 3-8 in tumor immunotherapy.
  10. 10. The use according to claim 9, wherein the covalent inhibitor derivative of a bioorthogonal group targets binding oncoproteins.

Description

Covalent inhibitor derivative of bio-orthogonal group, preparation method and application thereof Technical Field The invention relates to the technical field of chemical synthesis, in particular to a covalent inhibitor derivative of a bio-orthogonal group, a preparation method and application thereof. Background Cancer is clinically treated by using surgery, radiotherapy, cytotoxic chemotherapy, hormone therapy, targeted therapy, immunotherapy and the like as anticancer means singly or in combination according to tumor stage, resectability, biological characteristics, complications and overall patient status. Wherein, targeting therapy (such as small molecule signal transduction inhibitor, receptor tyrosine kinase antibody) aiming at specific mutation oncogene or downstream signal transduction cascade components thereof and immunotherapy of activating or enhancing specific recognition of human immune system and killing tumor cells can realize accurate killing of cancer cells, thereby deeply changing the treatment pattern and survival prognosis of cancer patients. However, with the penetration of clinical applications, both targeted and immunotherapy show respective limitations. Targeted therapy is limited by scarcity of patentable drug targets, and drug resistance is often generated by mechanisms such as target secondary mutation, bypass activation, phenotype transformation and the like. The immune therapy is limited due to the problems of insufficient overall response rate, frequent immune related adverse reactions and the like, and the deep mechanism of the immune therapy relates to tumor heterogeneity and immune editing. Neoantigens (neoantigen) are completely new peptide fragments encoded by gene mutations specific for tumor cells, different from the wild-type protein sequence, presented on the cell surface via the Major Histocompatibility Complex (MHC), specifically recognized by T cells and activating an anti-tumor immune response. The new antigen is used as a therapeutic target with great potential, is only expressed in tumor tissues, is not present in normal tissues, has high specificity and strong immunogenicity, has extremely low off-target risk, and is considered as an ideal target for realizing accurate immunotherapy. In the prior art, the new antigen therapy has the problems of limited new antigen prediction accuracy, immune escape and insufficient generation of new antigen specific T cells, and the problems seriously restrict the development of the new antigen therapy. Disclosure of Invention The invention aims to provide a covalent inhibitor derivative of a bio-orthogonal group, a preparation method and application thereof, and provides a new approach for solving the problems of target selection limitation and heterogeneity in tumor immunotherapy by designing and synthesizing a covalent inhibitor modified by the bio-orthogonal group to induce generation of tumor specific new antigens. In order to achieve the above purpose, the invention provides a covalent inhibitor derivative of bio-orthogonal groups, which is obtained by introducing polyethylene glycol with different chain lengths as fusion proteins by taking KRASG12C inhibitor sotorasib as a modifying molecule and connecting different bio-orthogonal groups to obtain sotorasib derivative; the bioorthogonal groups include dibenzo ring Xin Guian, alkynyl and tetrazinyl; the chain length of the polyethylene glycol is 4-16. Preferably, the sotorasib derivatives include sotorasib-PEG 4-alkynyl, sotorasib-PEG 5-alkynyl, sotorasib-PEG 4-tetrazine, sotorasib-PEG4-DBCO, sotorasib-PEG10-DBCO, sotorasib-PEG16-DBCO; the sotorasib-PEG 4-alkynyl has the structural formula: ; the sotorasib-PEG 5-alkynyl has the structural formula: ; The sotorasib-PEG 4-tetrazine has the structural formula: ; The sotorasib-PEG4-DBCO has the structural formula: ; The structural formulas of sotorasib-PEG10-DBCO and sotorasib-PEG16-DBCO are as follows: ; wherein n is 10 or 16. The invention also provides a preparation method of the covalent inhibitor derivative of the bio-orthogonal group, which comprises the following steps: S1, dissolving one or more of a KRASG12C inhibitor sotorasib, bromoacetic acid tert-butyl ester, potassium carbonate or 2-bromoethyl (methyl) carbamic acid tert-butyl ester in N, N-dimethylformamide according to a proportion, and reacting to obtain an intermediate; s2, sequentially adding dichloromethane and trifluoroacetic acid, and reacting to obtain an intermediate; S3, adding the intermediate obtained from the compounds a and S2 into an N, N-Dimethylformamide (DMF) solvent or a Dichloromethane (DCM) solvent, and reacting to obtain a target product; The compound a is a small molecular compound of PEG-connected bio-orthogonal group or amino-polyethylene glycol-tert-butyl propionate. Preferably, S3 further comprises adding a condensing agent, which is HATU (chinese name 2- (7-azobenzotriazole) -N, N' -tetramethylurea hexafluorophosphate), an organic ba