CN-121987811-A - Targeted killing conjugate

Abstract

The invention discloses a targeting killing conjugate, which at least comprises a targeting unit and a killing unit, wherein the units are connected through a connecting structure. The invention provides complete preparation, purification, quality control and various clinical administration schemes by taking AS1411 AS a targeting unit, LTX315 AS a killing unit and PEG2000 AS a connecting arm. The conjugate has strong targeting property, high safety, controllable preparation process and industrialized production, is suitable for targeting treatment of systemic and local focuses, and has good application prospect.

Inventors

• MA WENJIE

Assignees

• 马文杰

Dates

Publication Date

20260508

Application Date

20260401

Claims (1)

1. 1. A target killing conjugate is characterized by at least comprising a target unit and a killing unit, wherein the target unit can be combined with a target part, the killing unit can generate at least one of injury effect and killing effect on a target, and the units are connected through a connecting structure.

Description

Targeted killing conjugate Technical Field The invention relates to the technical field of biological medicines, in particular to a targeting killing conjugate. Background The clinical treatment of malignant tumor still faces the core problems of insufficient precision, obvious toxic and side effects, limited administration mode and the like, and the traditional chemotherapy drugs lack the tumor specific recognition capability, and damage normal tissues easily while killing tumor cells, so that patients have poor tolerance and poor prognosis. The targeted therapy is taken as a revolutionary direction of tumor therapy, and the targeted unit and the killing unit are coupled to realize the cooperation of accurate drug delivery and efficient killing, so that the targeted therapy becomes a research hot spot, wherein the aptamer-peptide conjugate has the advantages of high aptamer specificity and high polypeptide bioactivity, and has a wide application prospect. Nucleolin is highly expressed on the surfaces of various tumor cell membranes, is an ideal tumor targeting target, and AS1411 is used AS a nucleic acid aptamer for targeting nucleolin, can realize accurate identification of tumor cells by specifically combining nucleolin, has good safety and no obvious immunogenicity, has no killing activity, and can play a role in treatment by being coupled with a killing unit. LTX315 as a novel oncolytic polypeptide can realize high-efficiency killing by destroying the stability of tumor cell membranes, has extremely low toxicity to normal cells, and has proved curative effects in various tumor models. The existing aptamer-peptide conjugate has a plurality of technical bottlenecks that the connecting arm is unreasonable to select, so that the conjugate is poor in stability and water solubility, the preparation process is complex, the parameters are ambiguous, industrial mass production is difficult to realize, the administration mode is single, different focus types and positions cannot be adapted, the purity and bioactivity of the product are difficult to meet the medicinal requirements, and the clinical transformation of the product is limited. Therefore, the conjugate which has reasonable structural design, controllable preparation process and flexible administration mode and can realize accurate targeting and efficient killing is developed, the defects of the prior art are overcome, the requirements of industrial production and clinical application are met, and the technical problem to be solved in the field is urgent. Disclosure of Invention The invention aims to provide the targeted killing conjugate which has simple integral structure, definite targeting, stable killing effect, can adapt to various clinical administration modes and can meet the medication requirements of different clinical treatment scenes; the conjugate has simple structural design, realizes accurate targeting and efficient killing through the synergistic effect of the targeting unit, the killing unit and the connecting structure, and has stable biological activity; PEG2000 is used as a connecting arm, so that the water solubility is good, the steric hindrance is small, the stability and the targeting binding efficiency of the conjugate can be obviously improved, the immunogenicity is avoided, and the safety is high; The preparation process is mature and controllable, industrial mass production can be realized, and the purity of the product meets the medicinal requirement; Can support intravenous injection, drug-carrying support, local perfusion and other modes of administration, has wide application range and strong clinical practicality, and has higher medicinal value. Detailed Description In order to make the technical scheme of the invention clearer, the invention is further described in detail by combining specific embodiments. Examples The embodiment provides complete preparation, purification and clinical application schemes of the targeted killing conjugate, and the raw materials, reagents, process parameters and administration modes used are clear and practicable, so that the targeted killing conjugate meets the requirements of industrial production and clinical application. 1. Raw materials and reagents 1. A targeting unit: AS1411 aptamer with a sequence 5'-GGTGGTGGTGGTTGTGGTGGTGGTGG-3', modified by 5' end amino, and purity more than or equal to 98%; 2.a killing unit, namely LTX315 polypeptide, activated by C terminal carboxyl, and the purity is more than or equal to 95%; 3. The connection structure is a difunctional PEG connecting agent NH 2 -PEG2000-COOH with the molecular weight of 2000Da; 4. The solvent and buffer solution are sterile and asepsis ultrapure water, 0.01mol/L PBS (pH 7.4), chromatographic pure Dimethylformamide (DMF), EDC, HCl and HOBt which are all of medicinal grade; 5. Auxiliary materials are medical mannitol and sodium chloride, and stent coating matrix is medical PLGA. 2. Conjugate preparation process Dissolution