KR101327181B1 - Gene Delivery Complex Comprising Renal Cell-Specific Peptide and Polyester Amine and Composition Comprising The Same for Preventing or Treating of Renal Fibrosis. - Google Patents

Abstract

The present invention relates to gene carriers comprising renal cell specific peptides and polyesteramine polymers. The renal cell specific peptide-polyesteramine, a non-viral gene carrier according to the present invention, can provide significantly low cytotoxicity and high transfection efficiency and thus can be used as an effective gene carrier. In the case of using such a gene carrier, hepatocyte growth factor (HGF) gene, which is a therapeutic gene, can be efficiently delivered to a target site, and high renal expression efficiency in the kidney can be suppressed to suppress fibrosis. In addition, since creatinine and urea concentrations can be significantly lowered, they can be effectively used for preventing and treating renal fibrosis.

Description

Gene Delivery Complex Comprising Renal Cell Specific Peptides And Polyesteramine Polymers And Compositions For Preventing Or Treating Renal Fibrosis Including The Same. (Gene Delivery Complex Comprising Renal Cell-Specific Peptide and Polyester Amine and Composition Comprising The Same for Preventing or Treating of Renal Fibrosis.}

The present invention relates to a gene carrier comprising a renal cell specific peptide and a polyesteramine polymer, and a gene therapeutic agent for preventing or treating renal fibrosis using the same.

Gene therapy refers to a method of introducing a therapeutic gene into a patient's body to correct a defective gene or add a new function to a cell to treat or prevent cancer, infectious disease, genetic disease, and the like. Gene therapy can have superior selectivity compared to treatment with common drugs, and can have a rate and rate of treatment of a disease that is difficult to obtain with conventional therapies. Gene therapy does not cure the symptoms of a disease, but rather a way of treating and eliminating the cause of the disease.

As described above, a medicinal product or a medicine composed of cells into which a genetic material is introduced is administered to the human body for the purpose of treating a disease. Gene therapy products are based on molecular biology and recombinant DNA technology, which has seen remarkable growth over the last 50 years. In 1970, they began full-scale development, starting with the first clinical license for gene therapy products.

In order to be effective in gene therapy, it is necessary to develop a gene delivery technology that delivers a therapeutic gene to a desired target cell to obtain high expression efficiency. Vectors for gene transfer should be low or free of toxicity and capable of delivering genes to the cells of interest selectively and effectively.

These gene carriers can be largely divided into viral and non-viral, which is a viral gene composed of retro virus (RV), adeno virus (AV), and adeno associated viruses (AAV). Although the carrier is very excellent in expression rate and persistence, safety by immune response is pointed out as a problem.

In order to overcome the safety problems of such viral vectors, non-viral vectors have been developed that use various cationic polymers as gene carriers as an alternative to traditional viral vectors.

Cationic polymers bind to genes to form "cationic polymer-gene" complexes, which induce eukaryotic transfection. The specific mechanism is that the positive charge on the surface of the polymer-gene complex and the negative charge on the cell surface undergo the process of releasing the gene from the endosome to the cytoplasm after binding of the complex to the cell surface by electrical interaction. Specific examples of such cationic polymers include polyethyleneimine (PEI), polyamidoamine, polyL-lysine, polyaminoesters, cationic homopolymers such as polypropyleneimine and polyethylene glycol (PEG) -blocks. Cationic block copolymers such as -polyL-lysine and PEG-PEI; and the like.

The polymers condense therapeutic genes (e.g., DNA) into nano-sized particles and protect them from degradation, as well as protect the therapeutic genes from undesirable interactions, and deliver them into the cytoplasm and nucleus intracellularly. There is an advantage that can be strengthened. Among them, polyethyleneimine (PEI) is one of the most widely used non-viral vectors in the field of gene therapy research. Polyethylenimine (PEI) is known to effectively compress therapeutic genes (e.g. DNA) into colloidal particles, and has the advantage of rapidly releasing PEI / DNA complexes from endosomes.

As such, many cationic polymers have a low serum concentration in It is easy to be considered as a gene carrier because it shows high gene transfer efficiency in vitro , but unfortunately in It is very difficult to apply in vitro transfection directly to in vivo use, where the serum is present. This is because the transfection of the cationic polymer-gene complex is severely inhibited by various factors present in serum in vivo, and the influx of genes into cells in vivo is not smooth.

Polyethyleneimine (PEI), which is the most widely studied gene carrier, also has a significant decrease in transfection efficiency in vivo , and also has problems such as low expression efficiency of genes in vivo due to high cytotoxicity and low blood compatibility. have. Therefore, in gene therapy of cationic polymers in In vivo applications, there is an urgent need for the development of gene carriers that have low cytotoxicity and low transfection efficiency even in the presence of serum.

The present invention has been made in view of the above problems, and the present inventors have conducted extensive studies to prepare gene carriers having high biocompatibility, low cytotoxicity and low transfection efficiency even in the presence of serum. 'Peptide-polyesteramine copolymers' have been found useful in treating renal fibrosis and have led to the present invention.

The present invention provides gene carriers comprising renal cell specific peptides and polyesteramine polymers.

The present invention also provides a gene delivery complex for preventing or treating renal fibrosis in which a therapeutic gene is bound to a gene carrier comprising a renal cell-specific peptide and a polyesteramine polymer.

The present invention also provides a composition for preventing or treating renal fibrosis comprising the gene delivery complex as an active ingredient.

The renal cell specific peptide-polyesteramine, which is a non-viral gene carrier according to the present invention, can provide significantly low cytotoxicity and high transfection efficiency and thus can be used as an effective gene carrier. When using such a gene carrier, hepatocyte growth factor (HGF) gene, which is a therapeutic gene, can be efficiently delivered to a desired site, and high expression efficiency in the kidney can be suppressed, thereby inhibiting fibrosis. In addition, since creatinine and urea concentrations can be significantly lowered, they can be effectively used for preventing and treating renal fibrosis.

1 is a schematic diagram showing a synthesis process of a polyester amine (PEA) polymer according to the present invention.
2 shows a ¹ H NMR spectral analysis of a polyesteramine (PEA) polymer.
Figure 3 schematically shows the synthesis process of the renal cell-specific peptide-polyesteramine copolymer of the present invention.
Figure 4 shows the NMR analysis of the renal cell-specific peptide-polyesteramine copolymer of the present invention.
Figure 5 is a photograph confirming the renal cell-specific peptide-polyesteramine copolymer / DNA complex of the present invention formed according to the ratio of the amine group of the copolymer and the phosphate group of the DNA through electrophoresis.
Figure 6 is a graph measuring the cytotoxicity according to the concentration of the renal cell-specific peptide-polyesteramine copolymer of the present invention.
Figure 7 is a graph showing the measurement of gene expression in the kidney cells of the renal cell-specific peptide-polyesteramine copolymer / DNA complex of the present invention.
8 is a graph showing tissue-specific DNA gene expression in vivo of the renal cell-specific peptide-polyesteramine copolymer / DNA complex of the present invention.
9 is a photograph showing an in vivo renal fibrosis animal model using mice (left) and rats (rat, right).
10 is a photograph showing the expression level of GFP gene by treating the kidney cell specific peptide-polyesteramine copolymer / GFP complex of the present invention in a renal fibrosis model, where M represents mice and R represents rats.
Figure 11 is a graph showing the measurement of creatine concentration in the serum after treatment of the renal cell specific peptide-polyesteramine copolymer / HGF complex of the present invention in the renal fibrosis model.
Figure 12 is a graph showing the measurement of urea concentration in the serum treated with kidney cell specific peptide-polyesteramine copolymer / HGF complex of the present invention in the renal fibrosis model.
Figure 13 shows the analysis of renal tissue pathology through H & E staining after treating the renal cell specific peptide-polyesteramine copolymer / HGF complex of the present invention in a renal fibrosis model (PEP-PEA / GFP: renal cell specific). Peptide-polyesteramine copolymer / GFP complex, UUO: kidney fibrosis model, PEA / HGF: polyesteramine / HGF complex, PEP-PEA / HGF: kidney cell specific peptide-polyesteramine copolymer / HGF complex).
FIG. 14 shows the results of evaluating collagen generated in the tissue through sirius red staining after treating the peptide-polyesteramine copolymer / HGF complex of the present invention in a renal fibrosis model (PEP- PEA / GFP: renal cell specific peptide-polyesteramine copolymer / GFP complex, UUO: renal fibrosis model, PEA / HGF: polyesteramine / HGF complex, PEP-PEA / HGF: renal cell specific peptide-polyester Amine copolymer / HGF complex).

Hereinafter, the present invention will be described in more detail.

The present invention provides gene carriers comprising renal cell specific peptides and polyesteramine polymers.

In the present invention, the "renal cell specific peptide" refers to a peptide that can be specifically recognized by the receptor renal cells. In one embodiment of the present invention may be HOOC-ELRGDRAHW-NH₂.

The polyester amine (PEA) polymer can be synthesized by copolymerizing a low molecular weight polyethylenimine (PEI) and glycerol dimethacrylate (GDM) (FIG. 1).

The gene carrier of the present invention may be prepared by reacting a carboxyl group of a renal cell-specific peptide with an amine group of a polyesteramine polymer with an amide bond (FIG. 3).

The present invention also provides a gene delivery complex for preventing or treating renal fibrosis in which a therapeutic gene is bound to a gene carrier comprising a renal cell-specific peptide and a polyesteramine polymer.

The renal cell-specific peptide refers to a peptide that specifically binds to renal cells, preferably HOOC-ELRGDRAHW-NH₂.

Gene delivery complex for preventing or treating renal fibrosis of the present invention means a gene therapy agent that can be used for the purpose of preventing or treating renal fibrosis.

The gene delivery complex is a complex in which a therapeutic gene is bound to the gene delivery system of the present invention.

The therapeutic gene may be an HGF gene, and preferably, it comprises an artificial sequence DNA of SEQ ID NO: 1. The artificial sequence DNA is characterized by consisting of a total of 3679 nucleotide sequences.

The present invention also provides a composition for preventing or treating renal fibrosis comprising the gene delivery complex as an active ingredient.

The composition for preventing or treating renal fibrosis according to the present invention may be a pharmaceutical composition.

The pharmaceutical composition for preventing or treating renal fibrosis of the present invention can be prepared by mixing the gene delivery complex of the present invention with a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier can be any substrate commonly used in injections. Examples of suitable carriers are vegetable oils such as propylene glycol, polyethylene glycol, olive oil and corn oil, organic esters for injections such as gelatin and ethyloleate. The pharmaceutical composition for preventing or treating renal fibrosis of the present invention may further include a filler, an anticoagulant, a lubricant, a humectant, a perfume, an emulsifier, a preservative, and the like. Pharmaceutical compositions for the prophylaxis or treatment of renal fibrosis of the present invention can be formulated using methods well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. Injectables may also be prepared as solutions, suspensions or colloidal solutions by adding osmotic pressure regulators, pH regulators, vegetable oils such as sesame oil or soybean oil, or surfactants such as lecithin, nonionic surfactants, etc., in the usual manner. . Such injectables may be prepared in powder form, lyophilized form and dissolved before use.

The pharmaceutical composition for preventing or treating nephrotic fibrosis of the present invention can be conveniently administered parenterally, for example, by subcutaneous, transmuscular or transdermal injection. Direct delivery of the composition may be by injection, subcutaneously, intraperitoneally, intravenously, intratumorally or intramuscularly, or into the gaps of the tissue, in the solid phase, dissolved in a pre-sterilized substrate if necessary immediately before gene therapy. If used, or in the case of a liquid can be used as it is without additional treatment.

The pharmaceutical composition for preventing or treating nephrotic fibrosis of the present invention may be administered at a dose of 0.01 to 100 mg locally to a disease site of a patient based on an active ingredient in application to a patient. However, it is to be understood that the actual dosage of the active ingredient should be determined in light of several relevant factors such as the route of administration, the age, sex, weight of the patient, the severity of the disease, etc. and therefore the dosage should not be construed in any way for the scope of the invention. It is not limited.

Advantages and features of the present invention, and methods of accomplishing the same, will be apparent from and elucidated with reference to the embodiments described hereinafter in detail. However, the present invention is not limited to the embodiments disclosed below, but may be implemented in various forms, and only the present embodiments are intended to complete the disclosure of the present invention, and have ordinary skill in the art to which the present invention pertains. It is provided to fully inform the scope of the invention, and the invention is defined only by the scope of the claims.

[Example]

Example 1. Polyesteramine  ( PEA ) Polymer synthesis.

Reagents such as branched polyethyleneimine (PEI, 1.2 and 25 kDa), glycerol dimethacrylate (GDM) (Mn = 228.25 Da), and methanol were purchased from Sigma (St Louis, MO, USA).

Glycerol dimethacrylate and polyethyleneimine were dissolved separately in anhydrous methanol and the glycerol dimethacrylate solution was slowly added to the polyethyleneimine solution and mixed in three different stoichiometric GDM: PEI ratios (see Table 1).

division Reactant (wt%) Supply ratio
GMD: PEI
(Mol / Mol) Of PEI (a)
Composition (Mol%) Copolymer yield (%) Molecular weight (b) GDM PEI GDM / PEI-1.2
(1: 1) 228.25 1200 1: 1 56.78 37.72 7,630 GDM / PEI-1.2
(1: 2) 228.25 1200 1: 2 55.55 67.30 7,690 GDM / PEI-1.2
(1: 4) 228.25 1200 1: 4 68.44 55.74 7,700 a: ¹ H NMR measurement, b: GPC measurement

The reaction mixture was then heated at 60 ° C. for 24 hours with constant stirring. The reaction mixture was dialyzed with distilled water using a Spectra / Por membrane (molecular mass cutoff 3.5 kDa Spectrum Medical Industries, Inc., California) at 4 ° C. for 24 hours. Finally the polymer was lyophilised and stored at 0 ° C.

The reaction was carried out at an elevated temperature (60 ° C.) to increase the degree of crosslinking to increase the molecular weight of the synthesized polymer. Due to the nucleophilic nature of the amine group of polyethyleneimine it reacted with glycerol dimethacrylate end groups (FIG. 1).

Example 2. Polyesteramine  ( PEA Identification and Properties of Polymers

The composition of the synthesized polyesteramine was confirmed by ¹ H NMR spectroscopy. 2 shows a representative ¹ H NMR spectrum of GDM / PEI-1.2 (1: 1). The composition of the polyesteramine was found to vary depending on the stoichiometric feed ratio of the reactants. Polyesteramines have also been found to be soluble in highly polar solvents such as DMSO or water, but insoluble in some organic solvents such as THF.

The degree of polymer degradation was estimated by measuring the molecular weight reduction of PEA. The polymer dissolved in PBS (0.1 g / ml) was incubated in shake incubator (37 ° C., 100 rpm) and samples were taken at various time intervals. Subsequently, gel permeation chromatography was performed on the lyophilized sample to measure molecular weight.

In general, monomer structure, solvent selection, and reaction temperature affected the average molecular weight of the polymer, and their range was 2,000 to 50,000. However, the average molecular weight distribution of the synthesized polyesteramines has not been found to be very broad (close to polydispersity indices 2.0), suggesting that simply changing the reactant feed ratio suggests difficulty controlling the molecular weight ( See Table 1 above).

The polymer obtained above was dissolved in CDCl 3 at a concentration of 10 mg / ml to prepare a ¹ H NMR sample. NMR spectra were recorded using an Avance 500 minute photometer (Bruker, Germany). The molecular weight of the polymer was measured at a column temperature of 25 ° C. and a flow rate of 0.5 mL / min using a column (GPCusing Sodex OHpack SB-803 HQ column; Phenomenox, USA). Ammonium acetate (0.5M, pH 5.5) was used as mobile phase.

Biodegradability of the drug / gene delivery system does not mean that the carrier molecule collapses immediately after application, but rather slowly degrades to avoid long-term toxicity in the organism. In addition, the accumulation of undegraded and unreleased carriers causes cytotoxicity. The kinetics of ester bond degradation of the polyesteramines were investigated by measuring the decrease in molecular weight. As a result, the polyesteramines were hydrolyzed to form acids and alcohols, respectively, resulting in low molecular weight non-toxic byproducts. The half-life of GDM / PEI-1.2 (1: 1) was found to be 9-10 days, meaning that degradation occurred in a controlled manner.

Example 3. Kidney cell specific of the present invention Peptides - Polyesteramine  Synthesis of Copolymers.

Renal cell specific peptide-polyesteramine copolymer of the present invention The carboxyl group of the renal cell-specific peptide and the amine group of the polyesteramine were synthesized by reacting with an amide bond (FIG. 3). When preparing peptides, histamine was added to prepare a structure that can be identified by NMR. Renal cell specific peptides were purchased from Anigen.

The synthesized kidney cell specific peptide-polyesteramine copolymer is shown in FIG. 4. It was confirmed by NMR that 8.8 mol% of renal cell specific peptides were introduced into the polyesteramine.

Example 4. Kidney cell specific of the present invention Peptides - Polyesteramine  Copolymer / DNA Complex Preparation.

The renal cell-specific peptide-polyesteramine copolymer / DNA complex was formed by electrophoresis by controlling the ratio of the amine group of the polymer and the phosphate group of DNA to form an electrostatic action.

As a result, as shown in Figure 5, at a low weight ratio (weight ratio: 0.1 or 0.5), the composite is not properly formed, but it was confirmed that the composite is made by increasing the weight ratio to about 1.

In the following experimental example, the renal cell-specific peptide-polyesteramine copolymer of the present invention was simply referred to as' peptide-PEA ', and the renal cell-specific peptide-polyesteramine copolymer / DNA complex of the present invention was simply' peptide-PEA / DNA complex.

Experimental Example  1. of the present invention in renal cells peptide - PEA Cytotoxicity Assessment of.

Human kidney transformed cells (293T) were preserved in DMEM culture medium supplemented with 10% heat-inactivated FBS, penicillin / streptomycin. Cells were cultured under standard conditions in a 37 ° C. wet incubator containing 5% CO ₂ .

The cell viability of peptide-PEA was examined with a Cell Titer 96R Aqueous One Solution Cell Proliferation Kit (Promega). After 24 hours of incubation of cells loaded with an initial density of 1 × 10 ⁴ (293T) cells / well in a 0.2 well culture medium in a 96 well plate, the copolymer was concentrated at a concentration (5, 10, 20, 50 μg / ml). After 24 hours treatment, MTS assay was used to examine the cytotoxicity of the copolymer against 293T cell line. Polyethyleneimine 25K was used as a control.

As a result, as shown in Figure 6, it was found that the cytotoxicity gradually appeared as the concentration of the renal cell-specific peptide-PEA increases. In the case of PEI 25K, which is commonly known as a non-viral gene transfer material, the cell survival rate decreases rapidly as the concentration increases, but the peptide-PEA of the present invention showed a cell survival rate higher than 60% even at a high concentration of 20 µg / ml. Through this, peptide-PEA of the present invention was found to have a significantly higher cell viability than the control PEI 25K.

Experimental Example  2. of the present invention in renal cells peptide - PEA Of DNA  Investigation of gene expression of the complex.

Human kidney transformed cells (human kidney transformed cells, 293T) in 24 well plate 1 × 10 ⁵ cells incubated for 24 hours, and then treated with peptide-PEA / DNA complex of the present invention for 4 hours, the medium is changed and After 24 hours, the cells were lysed to confirm RLU (Relative Light Units) values using a luminometer, and RLU / mg protein values were calculated by checking protein concentrations using a BCA protein kit.

As a result, as shown in Figure 7, it was confirmed that the gene expression efficiency gradually increased in the peptide-PEA / DNA complex group as the weight ratio (weight ratio) increases. In addition, from the weight ratio 5, it was confirmed that the gene delivery efficiency was higher than that of the conventional PEI 25K and the weight ratio 10 was higher than the lipofectamine of the phospholipid series. In addition, higher gene expression efficiency was confirmed in the peptide-PEA / DNA complex group compared to the polyesteramine / DNA group.

Experimental Example  3. The peptide - PEA Of DNA  Composite in vivo In DNA  Gene Expression Survey.

To confirm whether the synthesized peptide-PEA / DNA complex shows gene expression efficiently in vivo, 30 μg of a reporter gene, GFP, is complexed with a renal cell-specific peptide-polyesteramine copolymer (peptide-PEA). The gene expression was examined after treatment to experimental animals by intravenous injection.

The animals were healthy and well-developed after 5 days of ICR mice and SD rats were purified for 3 days for evaluation of the gene expression test. Breeding conditions were the temperature 24 ± 3 ℃, relative humidity 50 ± 20% ventilation number 12-15 times / hour, lighting 12 hours (artificial lighting, 7 am to 7 pm). Breeding and testing of all experimental animals were conducted in accordance with the guidelines for management of experimental animals at Seoul National University. The purified mice received the peptide-PEA / GFP complex (GFP: 30 µg, total volume: 0.2 mL) through the tail vein and sacrificed mice after 48 hours, and expressed in major organs such as kidney, liver, heart and brain. The efficiency was confirmed.

As a result, as shown in FIG. 8, the group treated with peptide-PEA / GFP showed more GFP protein expression efficiency in renal tissue compared to the group treated with polyesteramine / GFP without binding to the renal cell-specific peptide. It was confirmed that it was shown. Through this, it was found that the present invention, in which the kidney-specific peptide is bound, works efficiently even in vivo.

Experimental Example  4. Construction of renal fibrosis model and renal fibrosis model peptide - PEA Of DNA  Composite DNA  Gene Expression Survey.

Example 4-1: Construction of a unilateral ureteral obstruction (UUO) in vivo

Renal fibrosis models in vivo were constructed with reference to Kinter et al., Kidney International (1999) 55, 1327-334. After anesthesia of the mice and rats, the urinary tract from the left kidney was ligated to inhibit renal function, thereby inducing renal fibrosis.

As a result, as shown in Figure 9, the right kidney (mouse and rat standard) compared to normal, the left kidney was induced renal fibrosis in two weeks, it was confirmed that the renal tissue is enlarged compared to the normal tissue even with the naked eye.

Example 4-2: Investigation of DNA Gene Expression of Peptide-PEA / DNA Complex in Kidney Fibrosis Model.

In order to confirm whether the synthesized peptide-PEA / DNA complex of the present invention exhibits efficient gene expression efficiency in the renal fibrosis model, 50 µg of the reporter gene, GFP gene, was formed with the peptide-PEA and then renal fibrosis. Model animals (for mice: GFP: 30 μg, total volume: 0.2 mL; for rats: GFP: 50 μg, total volume: 0.5 mL) were administered via tail vein and kidneys at the expense of mice and rats. GFP expression efficiency was confirmed in tissues.

As a result, as shown in Figure 10, the renal fibrosis model animal treated with the peptide-PEA / DNA complex of the present invention not only efficiently GFP protein expression in the right kidney tissue, but also GFP protein expression in the renal tissue fibrosis Was observed. This suggests that peptide-PEA efficiently delivers DNA genes to renal tissue.

Experimental Example  5. In the renal fibrosis model peptide - PEA Of HGF  Composite HGF  Gene Expression Survey.

Nanocomplexes (peptide-PEA / HGF) by combining the therapeutic HGF gene with peptide-PEA polymer in the renal fibrosis induction model (n = 4) based on the results of GFP gene expression of peptide-PEA / GFP in vivo After the formation of the rat (rat) by injection into the tail vein was confirmed the HGF gene expression efficiency. Peptide-PEA / HGF complex (HGF: 50 µg, total volume: 0.5 mL) was administered to the renal fibrosis model rat three times through the tail vein at three-day intervals from the day of fibrosis. rats were sacrificed to determine the efficiency of HGF gene expression in the kidney.

In the kidney function test, the blood was collected by cardiac collection, and the progress of kidney fibrosis was evaluated by measuring the levels of creatinine and urea in serum. As a control, sham control, HGF, PEA / HGF, peptide-PEA / HGF were used.

As a result, as shown in Figure 11 and 12, the group treated with peptide-PEA / HGF was confirmed that the concentration of creatinine and urea much lower than the group (UUO) induced kidney fibrosis.

Experimental Example  6. In the Kidney Fibrosis Model peptide - PEA Of HGF  Renal tissue pathology analysis after administration of the complex

Kidney histopathology was analyzed by H & E staining and sirius red staining after peptide-PEA / HGF complex administration in the renal fibrosis model. Rat kidney tissues treated with Sham control, Vector control (PEP-PEA / GFP), HGF, PEA / HGF, and PEP-PEA / HGF were identified by hematoxylin and eosin (H & E) staining, respectively.

As a result, as shown in FIG. 13, HGF-treated rat kidney tissues were identified by H & E staining, and renal fibrosis was induced in all groups by UUO procedure. It was confirmed that medulla thickness decreased as pelvis and papilla were pushed into medulla and cortex due to urethral obstruction, but PEA / HGF group and peptide-PEA / HGF group were weaker than control group. there was. As a result of the progression of renal fibrosis, it was confirmed that the index decreased compared to the control group. This suggests that the formation of fiber is much rarer in the group treated with PEA / HGF and peptide-PEA / HGF than the control UUO group.

Figure 14 shows the results of the evaluation of collagen generated in the tissue through the sirius red staining after administration of peptide-PEA / HGF in the renal fibrosis model, another control in the group treated with peptide-PEA / HGF Collagen formation is much rarer than in the group. The results showed similar results to H & E staining, and the peptide-PEA / HGF group showed a marked decrease in fibrous tissue compared to other groups.

Attach an electronic file to a sequence list

Claims (8)

A gene carrier comprising a copolymer of a polyesteramine polymer, polyethylenimine and glycerol dimethacrylate, and HOOC-ELRGDRAHW-NH ₂ , a renal cell specific peptide.
delete delete The method of claim 1, wherein the gene carrier comprises: a carboxyl group of HOOC-ELRGDRAHW-NH ₂ ; And an amine group of a copolymer of polyethyleneimine and glycerol dimethacrylate; and a gene carrier prepared by reacting an amide bond.
A gene transfer complex in which a hepatocyte growth factor (HGF) gene is combined as a therapeutic gene in a gene delivery system according to any one of claims 1 and 4.
delete The gene delivery complex according to claim 5, wherein the hepatocyte growth factor gene is a base of SEQ ID NO: 1.
Composition for preventing or treating renal fibrosis comprising the gene delivery complex according to claim 5.

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