Nuclide delivery system based on elastin-like polypeptide and preparation method thereof

Abstract

The invention belongs to the technical field of biological medicine, and provides an elastin-like polypeptide-based nuclide delivery system and a preparation method thereof, the elastin-like polypeptide-based nuclide delivery system comprises a carrier and radioactive iodine nuclide, the carrier is RGD-targeted amphiphilic elastin-like polypeptide, and the carrier is RGD-targeted amphiphilic elastin-like polypeptide; according to the nuclide delivery system based on the elastin-like polypeptide and the preparation method of the nuclide delivery system, the preparation process is simple, operation is easy and convenient, transportation is conducted in vivo in a micelle particle mode through the delivery system, the blood circulation time can be shortened, and tumor accumulation can be increased; by introducing the RGD sequence, the system can effectively target tumor cells, the tumor selectivity of radionuclides can be improved, and meanwhile, the toxicity to healthy tissues is reduced. The patent is subsidized by Key Technology and Industrial Demonstration for Green Biological Manufacturing of Medicinal Polypeptides, and the project number is 2021YFC2103900.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical technology, and specifically relates to a radionuclide delivery system based on elastin-like polypeptide and a preparation method thereof. Background Art

[0002] A radionuclide refers to an unstable atomic nucleus that can spontaneously emit rays (such as α, β rays, etc.) and can form a stable nuclide through decay. Utilizing the radiation particles and / or electromagnetic waves emitted by radionuclides is an effective method for cancer diagnosis and treatment, and thus the field of nuclear medicine has been developed. When a radionuclide is localized within a tumor, it can act as a therapeutic agent to destroy and eliminate exposed cancer cells; after introducing a radionuclide into the body, it can also serve as a diagnostic reagent. Based on the difference in the uptake of radionuclides by normal tissues and diseased tissues, images of the lesions can be obtained using imaging instruments. Usually, a class of radionuclides can be used for both diagnosis and treatment simultaneously, with one isotope serving as an imaging agent and the other as a therapeutic agent. With the continuous development and verification of new drugs, radionuclides have played an important role in tumor imaging and treatment.

[0003] However, radionuclides are highly efficient and common cytotoxic agents that can damage cells through ionizing radiation. Therefore, when the radionuclides fail to target the target cells, they will cause varying degrees of damage to the surrounding healthy cells, and radionuclides at therapeutic doses often cause certain adverse reactions. Summary of the Invention

[0004] In order to solve the above technical problems, the present invention provides a radionuclide delivery system based on elastin-like polypeptide and a preparation method thereof to solve the problems mentioned in the background art.

[0005] According to the first aspect of the present disclosure, a radionuclide delivery system based on elastin-like polypeptide is proposed, which includes a carrier and a radioactive iodine nuclide, and the carrier is an RGD-targeted amphiphilic elastin-like polypeptide.

[0006] Preferably, the particle size of the radionuclide delivery system based on elastin-like polypeptide is 50 - 200 nm, and the critical micelle temperature is 4 - 30 °C.

[0007] According to the second aspect of the present disclosure, a preparation method of a radionuclide delivery system based on elastin-like polypeptide is proposed, which includes the following steps:

[0008] S1. Prepare an RGD-targeted amphiphilic elastin-like polypeptide;

[0009] S2. Mix the RGD-targeted amphiphilic elastin-like polypeptide with a radioactive iodine salt, and add it to a pre-prepared Iodogen tube for reaction;

[0010] S3. Ultrafilter the reaction system in S2 to remove unreacted iodine, and obtain a radionuclide delivery system based on RGD-targeted amphiphilic elastin-like polypeptide.

[0011] Preferably, the theoretical loading molar ratio of the RGD-targeted amphiphilic elastin-like polypeptide to the radioactive iodine salt is 1:20 - 50.

[0012] Preferably, the content in the Iodogen tube is 1,3,4,6-tetrachloro-3a,6a-diphenylglycoluril.

[0013] Preferably, during the reaction in S2, the reaction temperature is 0 - 6°C and the pH value is 7 - 8.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] A radionuclide delivery system based on elastin-like polypeptide and its preparation method provided by the present invention have good biocompatibility, are biodegradable, and have a wide range of applications. On the one hand, the delivery system is transported in the form of micelle particles in the body, which can improve the blood circulation time and increase tumor accumulation. On the other hand, the introduction of the RGD sequence can enable the system to effectively target tumor cells, help improve the tumor selectivity of radionuclides, and at the same time reduce the toxicity to healthy tissues. And the delivery system can be applied to fields such as tumor radiotherapy and imaging. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 It is a schematic diagram of the electrophoresis result of ELP-RGD provided by the embodiment of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0017] The following further describes in detail the embodiments of the present invention in conjunction with the drawings. The following embodiments are used to illustrate the present invention, but cannot be used to limit the scope of the present invention.

[0018] As shown in the Figure 1 drawing:

[0019] Example 1: The present invention provides a radionuclide delivery system based on elastin-like polypeptide, including a carrier and a radioactive iodine nuclide. The radioactive iodine nuclide is iodine-125, and the carrier is an RGD-targeted amphiphilic elastin-like polypeptide.

[0020] The particle size of the radionuclide delivery system based on elastin-like polypeptide is 50 - 100 nm, and the critical micelle temperature is lower than 10°C.

[0021] A preparation method of a radionuclide delivery system based on elastin-like polypeptide includes the following steps:

[0022] S1. Preparation of the Iodogen tube: Dissolve 1,3,4,6 - tetrachloro - 3a,6a - diphenylglycoluril in dichloromethane, take an appropriate amount and add it to the bottom of a clamp - mouth bottle. Dry it with nitrogen, seal the bottle, evacuate the air, and store it in a - 20°C refrigerator for later use.

[0023] S2. Preparation of the RGD - targeted amphiphilic elastin - like polypeptide ELP - RGD;

[0024] S3. Add the ELP - RGD to be labeled to the PB buffer with a pH of 7.4, then add the Na125I solution and mix well. Add the mixture to the Iodogen tube and incubate for the reaction.

[0025] S4. After purifying the reaction solution through a Sephadex G25 chromatography column, a radionuclide delivery system based on ELP - RGD is obtained.

[0026] In S3, the reaction molar ratio of ELP - RGD to Na 125 I is 1:30, the incubation temperature is 4°C, and the incubation time is 25 min. By controlling the incubation temperature and time, the iodination efficiency of ELP - RGD is improved.

[0027] Example 2: A radionuclide delivery system based on an elastin - like polypeptide, comprising a carrier and a radioactive iodine radionuclide; the radioactive iodine radionuclide is iodine - 131; the carrier is an RGD - targeted amphiphilic elastin - like polypeptide.

[0028] The particle size of the radionuclide delivery system based on the elastin - like polypeptide is 50 - 100 nm, and the critical micelle temperature is lower than 10°C.

[0029] A preparation method of a radionuclide delivery system based on an elastin - like polypeptide, comprising the following steps:

[0030] S1. Preparation of the Iodogen tube: Dissolve 1,3,4,6 - tetrachloro - 3a,6a - diphenylglycoluril in dichloromethane, take an appropriate amount and add it to the bottom of a clamp - mouth bottle. Dry it with nitrogen, seal the bottle, evacuate the air, and store it in a - 20°C refrigerator for later use

[0031] S2. Preparation of the RGD - targeted amphiphilic elastin - like polypeptide ELP - RGD;

[0032] S3. Add the ELP - RGD to be labeled to the PB buffer with a pH of 7.4, then add the Na131I solution and mix well. Add the mixture to the Iodogen tube and incubate for the reaction.

[0033] S4. After purifying the reaction solution through a Sephadex G25 chromatography column, a radionuclide delivery system based on ELP - RGD is obtained.

[0034] In S3, the reaction molar ratio of ELP-RGD and Na 131 I is 1:25, the incubation temperature is 4 °C, and the incubation time is 30 min. By controlling the incubation temperature and time, the iodination efficiency of ELP-RGD is improved.

[0035] Example 3: A radionuclide delivery system based on elastin-like polypeptide, comprising a carrier and a radioactive iodine nuclide; the radioactive iodine nuclide is iodine-123, and the carrier is an RGD-targeted amphiphilic elastin-like polypeptide.

[0036] The particle size of the radionuclide delivery system based on elastin-like polypeptide is 100 - 200 nm, and the critical micelle temperature is lower than 10 °C.

[0037] A preparation method of a radionuclide delivery system based on elastin-like polypeptide, comprising the following steps:

[0038] S1. Prepare an iodogen tube: Dissolve 1,3,4,6-tetrachloro-3a,6a-diphenylglycouril in dichloromethane, take an appropriate amount and add it to the bottom of a crimp-neck bottle, dry it with nitrogen, seal the bottle, exhaust the air, and store it in a -20 °C refrigerator for later use.

[0039] S2. Prepare an RGD-targeted amphiphilic elastin-like polypeptide ELP-RGD.

[0040] S3. Add the ELP-RGD to be labeled in a PB buffer with a pH of 7.4, then add a Na123I solution and mix evenly. Add the mixture to the iodogen tube and incubate for reaction.

[0041] S4. After purifying the reaction solution through a Sephadex G25 chromatography column, a radionuclide delivery system based on ELP-RGD is obtained.

[0042] In S3, the reaction molar ratio of ELP-RGD and Na 123 I is 1:50, the incubation temperature is 4 °C, and the incubation time is 20 min. By controlling the incubation temperature and time, the iodination efficiency of ELP-RGD is improved.

[0043] Importantly, it should be noted that the structures and arrangements of the present application shown in multiple different exemplary embodiments are merely illustrative. Although only a few embodiments are described in detail in this disclosure, those who refer to this disclosure should easily understand that many modifications are possible without substantially departing from the novel teachings and advantages of the subject matter described in this application. Other substitutions, modifications, changes, and omissions can be made in the design, operating conditions, and arrangements of the exemplary embodiments without departing from the scope of the present invention. Therefore, the present invention is not limited to specific embodiments, but extends to various modifications that still fall within the scope of the appended claims.

[0044] In addition, in order to provide a concise description of the exemplary embodiments, all features of the actual embodiments may not be described (i.e., those features that are not relevant to the currently contemplated best mode of carrying out the invention or those features that are not relevant to implementing the invention).

[0045] It should be understood that in the development of any actual implementation, as in any engineering or design project, numerous specific implementation decisions may be made. Such development efforts may be complex and time-consuming, but for those of ordinary skill in the art who benefit from this disclosure, without undue experimentation, such development efforts will be a routine task of design, fabrication, and production.

[0046] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention may be modified or equivalently replaced without departing from the spirit and scope of the technical solutions of the present invention, and they should all be covered by the scope of the claims of the present invention.

Claims

1. A nuclide delivery system based on elastin-like polypeptide, characterized in that: The invention comprises a carrier and a radioactive iodine nuclide, wherein the carrier is an amphipathic elastin-like polypeptide targeted by RGD.

2. A nuclide delivery system based on elastin-like polypeptide according to claim 1, characterized in that: The particle size of the elastin-like polypeptide-based nuclide delivery system is 50-200 nm, and the critical micelle temperature is 4-30° C.

3. A method for preparing a nuclide delivery system based on an elastin-like polypeptide, applied to a nuclide delivery system based on an elastin-like polypeptide according to any one of claims 1 to 2, characterized in that: The steps include: S1. preparing an RGD-targeted amphiphilic elastin-like polypeptide; S2, mixing the RGD-targeted amphiphilic elastin-like polypeptide with radioactive iodine salt and adding the mixture into the pre-prepared Iodogen tube for reaction; S3, ultrafiltering the reaction system in S2 to remove unreacted iodine, and obtaining a nuclide delivery system based on RGD-targeted amphiphilic elastin-like polypeptide.

4. A method for preparing a nuclide delivery system based on elastin-like polypeptide according to claim 3, characterized in that: The theoretical loading molar ratio of the RGD-targeted amphiphilic elastin-like polypeptide to the radioiodine salt is 1:20-50.

5. A method for preparing a nuclide delivery system based on elastin-like polypeptide according to claim 3, characterized in that: The content of the Iodogen tube is 1,3,4,6-tetrachloro-3a,6a-diphenyl glycoluril.

6. A method for preparing a nuclide delivery system based on elastin-like polypeptide according to claim 3, characterized in that: During the S2 reaction, the reaction temperature is 0-6°C and the pH value is 7-8.

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