CN117551205A - Recombinant targeted ferritin-nano selenium hybrid compound and application thereof - Google Patents
Recombinant targeted ferritin-nano selenium hybrid compound and application thereof Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/04—Sulfur, selenium or tellurium; Compounds thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/0005—Vertebrate antigens
- A61K39/0011—Cancer antigens
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
- A61K2039/6031—Proteins
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/33—Fusion polypeptide fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
Abstract
The invention displays tumor targeting peptide at the amino end of caged protein by a genetic engineering method. Nanometer selenium is synthesized in the inner cavity of the cage-shaped protein in a bionic synthesis mode. The caged protein nano-selenium has good anti-tumor (solid tumor and blood tumor) effect, and particularly for leukemia treatment, the caged protein nano-selenium obviously induces leukemia cell differentiation and apoptosis. Meanwhile, the caged protein nano selenium can activate the tumor immune system and is used as a tumor vaccine. The caged protein nano-selenium prepared by the invention has important biomedical application potential for resisting solid tumors, inducing leukemia cell differentiation and apoptosis and activating tumor immune system.
Description
Technical Field
The invention belongs to the technical field of ferritin modification, and relates to a recombinant targeted ferritin-nano selenium hybrid compound and application thereof.
Background
Ferritin, a natural iron storage protein, is composed of 24-mers consisting of heavy and light chains. By utilizing the inner cavity of ferritin with the size of 8-12 nm, nano particles with uniform and dispersed sizes can be synthesized in a biomineralization mode. Targeting can be conferred by chemically or genetically displaying the targeting sequence on its surface. In 2007, ferroferric oxide nano enzyme is reported for the first time, and in 2 012, ferroferric oxide is loaded in the inner cavity of human heavy chain ferritin, and based on the characteristic that human heavy chain ferritin can be combined with tfR in a targeted manner, tumor visual diagnosis is realized. After that, there are studies on the use of ferritin nanoenzymes loaded with different metal elements for the treatment of various diseases. Ferritin has important application potential in the disease diagnosis and treatment process.
Selenium is an essential element in human body and has potential anti-tumor effect. Research has also shown that selenium has an immune activating function. In 2001, research on synthesizing red nano simple substance selenium is carried out, and nano selenium has good anti-tumor property and low toxicity. Selenium was used in leukemia treatment and was found to cause apoptosis in leukemia cells. Nano selenium is used for treating acute myeloid leukemia, and can cause apoptosis of acute myeloid leukemia cells.
However, no report has been made on the synthesis and application of ferritin-nanoselenium.
Disclosure of Invention
In order to overcome the defects in the prior art, the nano selenium is synthesized in the inner cavity of the ferritin in a bionic way, and the tumor targeting peptide is displayed on the surface of the ferritin by using a genetic engineering method, so that the targeting accurate treatment of tumors can be effectively realized.
In a first aspect, the invention provides a recombinant targeted ferritin comprising a targeting peptide displayed at the amino terminus of ferritin.
Ferritin "HFn" refers to any ferritin that may form a cage-like structure, which may be of natural origin, or may be a recombinantly expressed ferritin, or a mutant thereof, which may be derived from a prokaryote, a protozoa, a fungus, a plant or an animal, e.g. from a bacterium, a fungus, an insect, a reptile, an avian, an amphibian, a fish, a mammal, e.g. from a rodent, a ruminant, a non-human primate or a human, e.g. a mouse, a rat, a guinea pig, a canine, a cat, a cow, a horse, a sheep, a monkey, a gorilla, a human. Although the amino acid sequences of ferritin vary greatly from bacteria to humans, they are structurally similar and can form a protein shell structure.
In certain embodiments, the ferritin is a polypeptide having the nucleotide sequence of SEQ ID No.1.
In certain embodiments, the targeting peptide consists of 5-200 amino acids, preferably 5-15 amino acids.
In certain embodiments, the targeting peptide is a leukemia targeting peptide.
In certain embodiments, the nucleotide sequence of the recombinant targeted ferritin is SEQ ID No.2.
In certain embodiments, the amino acid sequence of the targeting peptide is CLL1 target targeting peptide C DLRSAAVC (SEQ ID No. 3) or VLA-4 target targeting peptide CPLDIDFYC (S EQ ID No. 4).
In a second aspect of the invention, there is provided a ferritin-nanoselenium hybrid complex for loading nanoselenium particles in the inner cavity of a recombinant targeted ferritin provided in the first aspect of the invention.
In certain embodiments, the loading ratio of recombinant targeted ferritin to selenium atoms is 1:10 to 1:1000, preferably 1:500 to 1:700.
In a third aspect, the invention provides an application of the recombinant targeted ferritin-nano-selenium hybrid compound in the second aspect in preparing a drug for targeted treatment of tumor.
In certain embodiments, the tumor comprises a solid tumor and a hematological tumor.
In certain embodiments, the hematological neoplasm is leukemia.
In certain embodiments, the leukemia is Acute Myeloid Leukemia (AML) or Acute Promyelocytic Leukemia (APL).
In a fourth aspect, the invention provides the use of a recombinant targeted ferritin-nanoselenium hybrid complex according to the second aspect of the invention in the preparation of a medicament for inducing differentiation and/or apoptosis of leukemic cells.
In a fifth aspect, the present invention provides an immune activator, wherein the immune activator contains the recombinant targeted ferritin-nano-selenium hybrid complex according to the second aspect as an active ingredient.
Compared with the prior art, the invention displays the tumor targeting peptide at the amino end of human heavy chain ferritin (HFn) for the first time by a genetic engineering method, and synthesizes nano selenium in the ferritin cavity by a bionic synthesis mode. The ferritin nano-selenium has good anti-tumor (solid tumor and blood tumor) effect, and particularly for leukemia treatment, the ferritin nano-selenium obviously induces leukemia cell differentiation and apoptosis. Ferritin nano-selenium (AE-HFn-Se) can be used as a Histone Deacetylase (HDAC) inhibitor for treating tumors (solid tumors and hematological tumors). Meanwhile, the ferritin nano-selenium can activate the tumor immune system and is used as a tumor vaccine. The ferritin nano-selenium prepared by the invention has important biomedical application potential for resisting solid tumor and inducing leukemia cell differentiation and apoptosis and activating tumor immune system.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments will be briefly described below.
FIG. 1 AE-HFn recombinant ferritin purification and characterization results. Wherein, graph A is SD S-PAGE to verify AE-HFn single subunit molecular weight; panel B is SEC showing the size of AE-HFn assembled into ferritin; panel C shows that DLS shows an AE-HFn average radius size of about 7.4nm; and the graph D shows that AE-HFn is spherical and has better monodispersity by a transmission electron microscope.
FIG. 2 AE-HFn-Se characterization. Wherein, the graph A is TEM AE-HFn-Se negative staining result, the graph B is TEM AE-HFn-Se non-staining result, the graph C is AE-HFn-S e high resolution transmission electron microscope graph, and the graph D is AE-HFn-Se binding energy absorption peak detected by XPS. Panel E shows the selenium content of AE-HFn-Se measured by ICP-MS.
FIG. 3 sensitivity test of tumor cells (leukemia and solid tumor) to AE-HFn-Se, wherein FIG. A shows the results of sensitivity test of leukemia cell lines Kasumi-1, K562, U937, and FIG. B shows the results of sensitivity test of solid tumor cell lines A549, hepG2, U87 to AE-HFn-Se.
FIG. 4 shows the results of targeting detection of leukemia cells by ferritin nanoselenium. Wherein, the graph A is siRNA knockdown ITGA, the graph B is the targeting detection of AE-HFn-FITC to ITGA knockdown cell Kasu mi-1, and the graph C is the targeting detection of AE-HFn-FITC to CD 71.
FIG. 5 detection of differentiation of ferritin nanoselenium-induced leukemia cells. Wherein, FIG. A is Swiss staining, FIG. B is the effect of AE-HFn-Se on the expression of CD11B in Kasumi-1 cells, and FIG. C is the effect of AE-HFn-Se on the expression of CD14 in Kasumi-1 cells.
FIG. 6 shows the results of detection of apoptosis of ferritin nano-selenium-induced leukemia cells. Wherein, the graph A is that the flow cytometry detects that AE-HFn-Se induces Kasumi-1 cells to undergo apoptosis in a time-dependent and dose-dependent manner, and the graph B is that WB detects that AE-HFn-Se induces clear caspase3 protein expression level to be increased.
FIG. 7 shows the results of detection of the effect of ferritin nanoselenium on leukemia H3K9 acetylation. Wherein, the graph A shows the effect of AE-HFn-Se on H3K9 acetylation of Kasumi-1 cells, and the graph B shows the effect of AE-HFn-Se on H3K9 acetylation of NB4 cells.
FIG. 8 shows a graph of change in CD80 expression of HFn-Se-induced dendritic cells.
Detailed Description
General overview
The invention relates to a protein-nano selenium hybrid compound which mainly contains targeted protein and nano selenium. The protein-nano selenium hybrid compound can be used for treating tumors (solid tumors and blood tumors) and can be used as an immune vaccine. Specifically, the invention displays tumor targeting peptides at the amino terminus of human heavy chain ferritin (HFn) by genetic engineering methods. Nanometer selenium is synthesized in the inner cavity of ferritin in a bionic synthesis mode. The ferritin nano-selenium has good anti-tumor (solid tumor and blood tumor) effect, and particularly for leukemia treatment, the ferritin nano-selenium obviously induces leukemia cell differentiation and apoptosis. Ferritin nano-selenium (AE-HFn-Se) can be used as a Histone Deacetylase (HDAC) inhibitor for treating tumors (solid tumors and hematological tumors). Meanwhile, the ferritin nano-selenium can activate the tumor immune system and is used as a tumor vaccine. The ferritin nano-selenium prepared by the invention has important biomedical application potential for resisting solid tumor and inducing leukemia cell differentiation and apoptosis and activating tumor immune system.
Detailed Description
Embodiments of the technical scheme of the present invention will be described in detail below. The following examples are only for more clearly illustrating the technical aspects of the present invention, and thus are merely examples, which should not be construed as limiting the scope of the present invention. It is noted that unless otherwise indicated, technical or scientific terms used herein should be given the ordinary meaning as understood by one of ordinary skill in the art to which this invention pertains.
EXAMPLE 1 recombinant engineering human heavy chain ferritin purification and characterization
Construction of AE (AML 1-ETO short) -HFn plasmid
AE is AML1-ETO for short, nine amino acid gene sequences (which are mainly targeted to bind AML1-ETO leukemia, AE for short) are displayed at the amino terminal of HFn complete gene sequence (SEQ ID NO. 1), a recombinant HFn sequence (AE-HFn) with targeting property is obtained, the sequence is shown as SEQ ID NO.2 (wherein, the bolded italic base sequence is a targeting peptide sequence and the underlined base sequence is HFn complete gene sequence), ndeI/BamHI restriction enzyme is selected as an enzyme cleavage site, and the recombinant HFn sequence is constructed on pET22b plasmid and is constructed by Shanghai Jieli.
CATATGTGTCCGTTAGATATTGATTTTTATTGTGGTGGCGGT GGCTCTATGACCACCGCAAGTACCT CACAGGTGCGCCAGAATTATCATCAGGATAGTGAAGCAGCAATTAATCGTCAGATTAATTTAGAACTGTATGCAAG CTATGTGTATCTGAGTATGAGCTATTATTTTGATCGCGATGATGTTGCCCTGAAAAATTTTGCCAAATATTTTCTG CATCAGTCTCATGAAGAACGCGAACATGCCGAAAAACTGATGAAATTACAGAATCAGCGTGGCGGTCGCATTTTTC TCCAAGATATTAAAAAACCGGATTGTGATGATTGGGAAAGCGGCCTGAATGCAATGGAATGTGCCTTACATCTGGA AAAAAATGTTAATCAGTCACTGCTGGAACTGCATAAACTGGCAACCGATAAAAATGATCCGCATCTGTGTGATTTT ATTGAAACCCATTATCTGAATGAACAGGTTAAAGCCATTAAAGAACTGGGCGATCATGTTACCAATTTACGTAAAA TGGGCGCCCCGGAAAGTGGCTTAGCCGAATATCTGTTTGATAAACATACCTTAGGCGATAGCGATAATGAATCTTA AGGATCC(SEQ ID NO.2)。
AE-HFn protein purification
E.coli BL21 (TransGen) expression strain was transformed with AE-HFn, and plates were plated with ampicillin L B, and the monoclonal was picked and shaken overnight. Then, ampicillin-resistant LB medium was added, and after extensive culture, the bacterial suspension was collected by centrifugation at 6000rpm and resuspended in 50mM Tris, pH 8.0. After resuspension, high pressure homogenization and disruption was performed, followed by centrifugation at 12000rpm for 20min, then heat treatment at 72℃for 15min, centrifugation at 12000rpm for 40min, collection of supernatant, filtration through a 0.22 um filter head, and sodium chloride supplementation to 0.75mM. The AE-HFn was then purified by a hydrophobic column, concentrated after purification, and further purified by separation using a Superdex200 molecular sieve. The AE-HFn protein concentration was determined using a BCA protein assay kit with bovine serum albumin as standard.
AE-HFn protein characterization
The size of the AE-HFn single subunit molecular weight after recombinant expression was verified by SDS-PAGE. Superdex200 molecular sieves and transmission electron microscopy examined AE-HFn assembly.
As a result, as shown in FIG. 1, the purified AE-HFn was found to have a larger molecular weight of the AE-HFn single subunit than HFn as verified by SDS-PAGE (A in FIG. 1). SEC showed that AE-H Fn peaked between 50-75ml, indicating that AE-HFn assembled into ferritin size (B in FIG. 1). DLS shows an AE-HFn average radius size of about 7.4nm (C in FIG. 1). Transmission electron microscopy showed AE-HFn to be spherical and well monodisperse (D in FIG. 1).
Example 2 Synthesis and characterization of ferritin nanoselenium
1. Synthesis of ferritin nano-selenium
Adding 2mM sodium selenite into 50mM tris-HCL 0.15M sodium chloride solution containing 1 μm AE-HFn, standing at room temperature for 3 hr, adding sodium borohydride, reducing for 1 hr, centrifuging at 10000 rpm for 15min, and adding Hiprep TM 26/10 Desantng was desalted and then purified using Superdex200 (10/300GL,GE Healthcare).
2. Ferritin nanoselenium characterization
AE-HFn-Se assembly and selenium nanoparticles were detected by TE M negative staining and non-negative staining, respectively, in PBS solution, ph=7.4, prepared at 0.1mg/ml AE-HFn-Se. 0.1mg/ml AE-HFn-Se is prepared, and in PBS solution, the content and nano-selenium lattice spacing are detected by ICP-MS and high-resolution transmission electron microscope. 50mg/ml AE-HFn-Se was replaced with water in PBS solution, lyophilized to a powder, sent to a middle laboratory for XPS.
The results are shown in FIG. 2, wherein the synthesized AE-HFn-Se is subjected to Hiprep TM 26/10Desa multiple ng was desalted and then purified by superdex200 (10/300GL,GE Healthcare) to be separated and colored red. TEM negative staining shows that AE-HFn-Se has a spherical shell structure and good monodispersity. TEM does not negatively stain, showing AE-HFn-Se contains nanoparticles about 4.5nm in size. High resolution transmission electron microscopy showed that the nanoparticles in AE-HFn-Se had a lattice spacing of about 0.216 nm. XPS shows a significant absorption peak at a binding energy of 55.3eV, indicating that the XPS is zero-valent nano-selenium. ICP-MS measures selenium content in AE-HFn-Se, each AE-HFn containing about 530 selenium atoms.
Example 3 sensitivity of tumor cells (leukemia and solid tumor) to ferritin nanoselenium
The sensitivity of leukemia cell lines (Kasumi-1, K562, U937) and solid tumor cell lines (A549, hepG2, U8 7) to AE-HFn-Se was examined separately using a cell proliferation-toxicity assay kit (CCK 8), and the half lethal dose (IC 50) was calculated.
As shown in FIG. 3, the tumor cells (leukemia and solid tumor) had a certain sensitivity to AE-HFn-Se, the leukemia cells showed a better sensitivity to AE-HFn-Se, and the leukemia cells Kasumi-1 were more sensitive to AE-HFn-Se.
Example 4 detection of leukemia cell targeting by ferritin nanoselenium
VLA-4 targeting detection: ITGA is taken as a subunit of a leukemia target VLA-4, and a targeting peptide displayed at the amino end of ferritin is mainly combined with ITGA, so that in order to verify the targeting of AE-HFn, siRNA is utilized to electroknock down ITGA, and the targeting of AE-HFn-FITC to Ka sumi-1 cells is detected.
CD71 targeting assay: HFn has better targeting specificity to CD71, and we detect Kasumi-1 surface CD71 through HFn-FITC.
The results are shown in FIG. 4, in which AE-HFn-FITC has reduced targeting binding to the ITGA knockdown group as compared to the control group. AE-HFn proved to be targeted to VLA-4. Meanwhile, the HFn can inhibit the binding specificity of AE-HFn to Kasumi-1, and the AE-HFn has targeting to CD7 1.
EXAMPLE 5 ferritin nanoselenium induces leukemia cell differentiation
Swiss staining: kasumi-1 cells were induced with 90ug/ml AE-HFn-Se, and the cells were changed every two days, and the cell morphology and nuclear changes were observed after continuous induction for 4 days.
Flow cytometry: kasumi-1 cells were induced with 30ug/ml, 90ug/ml, 180ug/ml AE-HFn-Se, and the cell surface molecules CD11b and CD14 were examined at 24h, 48h, and 72h, respectively.
The results are shown in FIG. 5, which shows that AE-HFn-Se induces differentiation of Kasumi-1 cells by Swiss staining. AE-HFn-Se induces a dose and time dependent up-regulation of the Kasumi-1 cell surface molecules CD11b, CD 14.
EXAMPLE 6 ferritin nanoselenium induces leukemia cell apoptosis
Flow cytometry: kasumi-1 cells were induced with 30ug/ml, 90ug/ml, 180ug/ml AE-HFn-Se, respectively, and double staining of Annexin V-FIT C and PI was performed at 24h, 48h, and 72h, to detect apoptosis of Kasumi-1 cells.
Western Blot: ka sumi-1 cells were induced with 90ug/ml and 180ug/ml AE-HFn-Se, respectively, and the clear caspase3 protein expression level was detected at 24 hours and 48 hours.
The results are shown in FIG. 6, which shows that AE-HFn-Se induces apoptosis of Kasumi-1 cells in a time-and dose-dependent manner. WB assay found that AE-HFn-Se induced an increase in the expression level of C-leaved caspase3 protein.
Example 7 ferritin nanoselenium inhibits histone deacetylase activity, leading to histone H3 acetylation
Kasumi-1 and NB4 cells were stimulated with different concentrations of AE-HFn-Se (60 ug/ml, 180ug/ml, 270 ug/ml), respectively, and after 48H, the effect of AE-HFn-Se on the acetylation levels of Kasumi-1 and NB4 cytohistones H3K9 was examined, respectively, by Western Blot experiments. The results are shown in FIG. 7, where AE-HFn-Se clearly caused a dose-dependent up-regulation of the acetylation levels of H3K9 in Kasumi-1 and NB4 cells.
EXAMPLE 8 iron protein nanoselenium-induced dendritic cell maturation
After DC2.4 was stimulated with HFn-Se (20 ug/ml) for 24 hours, the change in DC2.4 CD80 expression of mouse dendritic cells induced by HFn-Se was examined by flow cytometry to determine whether dendritic cell maturation could be induced. The results are shown in FIG. 8, in which HFn-Se induces up-regulation of DC2.4 CD80 expression in mouse dendritic cells, which can induce differentiation and maturation of mouse dendritic cells.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention, and are intended to be included within the scope of the appended claims and description.
Sequence listing
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Claims (14)
1.A recombinant targeted ferritin comprising a targeting peptide displayed at the amino terminus of ferritin.
2. The recombinant targeted ferritin according to claim 1, wherein the ferritin nucleotide sequence is SEQ ID No.1.
3. Recombinant targeted ferritin according to claim 1, wherein the targeting peptide consists of 5-200 amino acids, preferably 5-15 amino acids.
4. The recombinant targeted ferritin according to claim 1, wherein the targeting peptide is a leukemia targeting peptide.
5. The recombinant targeted ferritin according to claim 1, wherein the amino acid sequence of the targeting peptide is CLL1 target targeting peptide CDLRSAAVC or VLA-4 target targeting peptide CPLDIDFYC.
6. The recombinant targeted ferritin according to claim 5, wherein the nucleotide sequence of the recombinant targeted ferritin is SEQ ID No.2.
7. A recombinant targeted ferritin-nanoselenium hybrid complex wherein nanoselenium particles are loaded in the recombinant targeted ferritin lumen of any one of claims 1-6.
8. The recombinant targeted ferritin-nanoseleno hybrid complex according to claim 7, wherein the loading ratio of recombinant targeted ferritin to selenium atoms is 1:10 to 1:1000, preferably 1:500 to 1:700.
9. Use of the recombinant targeted ferritin-nano-selenium hybrid complex according to any of claims 7-8 in the preparation of a medicament for targeted treatment of tumors.
10. The use of claim 9, wherein the tumor comprises a solid tumor and a hematological tumor.
11. The use according to claim 10, wherein the hematological neoplasm is leukemia.
12. The use according to claim 11, wherein the leukemia is Acute Myeloid Leukemia (AML) or Acute Promyelocytic Leukemia (APL).
13. Use of a recombinant targeted ferritin-nanoseleno hybrid complex according to any one of claims 7-8 in the preparation of a medicament for inducing leukemia cell differentiation and/or apoptosis.
14. An immune activator, characterized in that the immune activator contains the recombinant targeted ferritin-nano-selenium hybrid complex as an active ingredient.
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| PCT/CN2022/086284 WO2023164991A1 (en) | 2022-03-04 | 2022-04-12 | Recombinant targeting ferritin-nano-selenium hybrid complex and use thereof |
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