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KR-20260065471-A - TRANSDERMAL DELIVERY OF GENETIC MATERIAL AND COMPOSITION COMPRISING SAME FOR PREVENTING OR TREATING CANCER

KR20260065471AKR 20260065471 AKR20260065471 AKR 20260065471AKR-20260065471-A

Abstract

The present invention relates to a genetic material delivery vehicle for transdermal delivery and a composition for preventing or treating cancer containing the same. The genetic material delivery vehicle according to the present invention is coated with hyaluronic acid (HA) on cationic solid lipid nanoparticles (CSLNs) that carry angiogenesis-inhibiting siVEGF, thereby promoting transdermal delivery of genetic material and targeting cancer cells. It has an appropriate nano-size distribution and minimal non-specific cytotoxicity, and can be used as a novel drug delivery vehicle for transdermal targeted cancer therapy.

Inventors

  • 한세광
  • 피아오, 정위
  • 김문구

Assignees

  • 포항공과대학교 산학협력단

Dates

Publication Date
20260508
Application Date
20250312
Priority Date
20241031

Claims (17)

  1. cationic solid lipid nanoparticles with a core-shell structure; and A coating layer comprising hyaluronic acid located on the above-mentioned cationic solid lipid nanoparticles; comprising The above core comprises cholesteryl ester and triglyceride, and The above shell is a genetic material carrier comprising cholesterol, fusion-inducible lipids, cationic lipids, and lipid-PEG (polyethylene glycol) conjugates.
  2. In paragraph 1, The above-mentioned genetic material carrier is a genetic material carrier that delivers genetic material through the skin.
  3. In paragraph 1, The above genetic material delivery vehicle further comprises genetic material, and A dielectric material carrier in which the dielectric material is located between the shell of the cationic solid lipid nanoparticle and the coating layer, and is bonded by electrostatic interaction with the cationic lipid of the shell.
  4. In paragraph 3, A genetic material carrier comprising one or more selected from the group consisting of small interfering ribonucleic acid (siRNA), ribosomal ribonucleic acid (rRNA), ribonucleic acid (RNA), deoxyribonucleic acid (DNA), complementary deoxyribonucleic acid (cDNA), aptamer, messenger ribonucleic acid (mRNA), transfer ribonucleic acid (tRNA), and antisense oligodeoxynucleotide (AS-ODN).
  5. In paragraph 3, A genetic material carrier in which the weight ratio (genetic material/cationic solid lipid nanoparticles) of the above-mentioned cationic solid lipid nanoparticles and the above-mentioned genetic material is 1 to 50.
  6. In paragraph 1, The above hyaluronic acid is a genetic material carrier having a molecular weight of 5 to 200 kDa.
  7. In paragraph 1, The above-mentioned cholesteryl ester is a genetic material carrier in which a saturated or unsaturated fatty acid having 10 to 24 carbon atoms is ester-bonded to cholesterol.
  8. In paragraph 1, A genetic material carrier in which the above triglyceride is triolein.
  9. In paragraph 1, The above fusion-inducing lipids are dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), egg phosphatidylcholine (EPC), distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylethanolamine (DSPE), and phosphatidylethanolamine A genetic material carrier comprising one or more selected from the group consisting of phosphatidylethanolamine (PE), dipalmitoylphosphatidylethanolamine, 1,2-dioleyl-sn-glycero-3-phosphoethanolamine, 1-palmitoyl-2-oleyl-sn-glycero-3-phosphoethanolamine (POPE), 1-palmitoyl-2-oleyl-sn-glycero-3-phosphocholine (POPC), 1,2-dioleyl-sn-glycero-3-[phospho-L-serine] (DOPS), and 1,2-dioleyl-sn-glycero-3-[phospho-L-serine].
  10. In paragraph 1, The above cationic lipids are 3-beta[N-(N',N'-dimethylaminoethane)carbamoyl]cholesterol (DC-cholesterol), 3-beta-[N-(N',N',N'-trimethylaminoethane)carbamoyl]cholesterol (TC-cholesterol), 3-beta[N-(N'-monomethylaminoethane)carbamoyl]cholesterol (MC-cholesterol), 3-beta[N-(aminoethane)carbamoyl]cholesterol (AC-cholesterol), N-(N'-aminoethane)carbamoylpropanoic tocopherol (AC-tocopherol), N-(N'-methylaminoethane)carbamoylpropanoic tocopherol (MC-tocopherol), N,N-dioleyl-N,N-dimethylammonium chloride (DODAC), N,N-Distearyl-N,N-Dimethylammonium Bromide (DDAB), N-(1-(2,3-Dioleoyloxy)propyl-N,N,N-Trimethylammonium Chloride (DOTAP), N,N-Dimethyl-(2,3-Dioleoyloxy)propylamine (DODMA), N-(1-(2,3-Dioleyl)propyl)-N,N,N-Trimethylammonium Chloride (DOTMA), 1,2-Dioleyl-3-Dimethylammonium-Propane (DODAP), 1,2-Dioleylcarbamyl-3-Dimethylammonium-Propane (DOCDAP), 1,2-Dilineoyl-3-Dimethylammonium-Propane (Dilineoyl-3-Dimethylammonium-propane, DLINDAP), Dioleoyloxy-N-[2-spermincarboxamido)ethyl}-N,N-dimethyl-1-propaneamulanttrifluoroacetate (DOSPA), dioctadecylamidoglycyl spermine (DOGS), 1,2-dimyristriloxypropyl-3-dimethyl-hydroxyethyl ammonium bromide (DMRIE), 3-dimethylamino-2-(cholest-5-en-3-beta-oxybutane-4-oxy)-1-(cis,cis-9,12-octadecadienoxy)propane (CLinDMA), 2-[5'-(cholest-5-en-3-beta-oxy)-3'-oxapentoxy]-3-damethyl-1-(cis,cis-9',12'-octadecadienoxy)propane (CpLinDMA), N,N-dimethyl-3,4-dioleyloxybenzylamine A genetic material carrier comprising one or more selected from the group consisting of (DMOBA), 1,2-N,N'-dioleylcarbamyl-3-dimethylaminopropane (DOcarbDAP), 1,2-diacyl-3-trimethylammonium-propane (TAP), 1,2-diacyl-3-dimethylammonium-propane (DAP), 1,2-di-O-octadeceyl-3-trimethylammonium propane, and 1,2-dioleyl-3-trimethylammonium propane.
  11. In paragraph 1, A genetic material delivery system comprising, based on 100 weight% of total cationic solid lipid nanoparticles, 30 to 60 weight% of the cholesteryl ester, 0.1 to 10 weight% of the triglyceride, 5 to 20 weight% of the cholesterol, 5 to 30 weight% of the fusion-inducing lipid, 10 to 50 weight% of the cationic lipid, and 0.01 to 1 weight% of the lipid-PEG conjugate.
  12. In paragraph 1, A dielectric material carrier having a content ratio between the core and shell of 30:70 to 70:30 by weight.
  13. In paragraph 1, A genetic material carrier having an average particle size of 30 to 500 nm.
  14. In paragraph 1, A genetic material carrier that exhibits a zeta potential of -30 to -5 mv.
  15. Core-shell structured cationic solid lipid nanoparticles; A coating layer containing hyaluronic acid located on the above positively charged solid lipid nanoparticles; and A dielectric material located between the shell of the positively charged solid lipid nanoparticle and the coating layer; comprising The above core comprises cholesteryl ester and triglyceride, and The above shell comprises cholesterol, fusion-inducing lipids, cationic lipids, and lipid-PEG (polyethylene glycol) conjugates, and A pharmaceutical composition for the prevention or treatment of cancer, wherein the above-mentioned genetic material is bonded to the cationic lipid of the shell by electrostatic interaction.
  16. In paragraph 15, The above composition is a pharmaceutical composition that delivers the genetic material through the skin.
  17. In paragraph 15, A pharmaceutical composition wherein the above cancer is any one selected from the group consisting of skin cancer, cutaneous or intraocular melanoma, colorectal cancer, breast cancer, brain cancer, neurocancer, lung cancer, small cell lung cancer, stomach cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, head or neck cancer, uterine cancer, ovarian cancer, rectal cancer, pro-anal cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal cell carcinoma, renopelvic carcinoma, CNS tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma and pituitary adenoma.

Description

Transdermal delivery of genetic material and composition for preventing or treating cancer comprising the same The present invention relates to a genetic material delivery vehicle for transdermal delivery and a composition for preventing or treating cancer containing the same. More specifically, it relates to a novel siRNA delivery system that forms a stable nanoscale complex capable of targeted delivery to skin cancer tissue by coating hyaluronic acid (HA) onto cationic solid lipid nanoparticles (CSLNs) carrying angiogenesis-inhibiting siVEGF. RNA interference (RNAi) is a cellular defense mechanism that sequence-specifically degrades foreign RNA. Short interfering RNA (siRNA) is a form of RNAi that binds to target RNA, cleaves and degrades it, and consequently downregulates the expression of target proteins. This mechanism is receiving significant attention in therapeutic applications due to its ability to downregulate specific proteins. To date, the FDA has approved six siRNA drugs (Patisiran (Onpattro), Givosiran (Givlaari), Lumasiran (Oxlumo), Inclisiran (Leqvio), Nedosiran (DCR-PHXC), and Vutrisiran (Amvutta)), most of which target the liver. These developments highlight the promising potential of siRNA for treating various diseases. However, the six approved drugs are limited to liver-targeted delivery because they primarily utilize GalNAc as a delivery carrier. While GalNAc has successfully delivered siRNA to hepatocytes, challenges remain, such as difficulties in targeting other organs, limited endocytotic escape, hepatotoxicity in non-target organs, and extensive enzymatic and systemic clearance. Currently, several other drugs are in Phase II and III clinical trials, primarily targeting organs such as the liver, eyes, lungs, kidneys, brain, and skin. However, since most are initially delivered in RNA form, the need to explore alternative delivery systems, such as lipid nanoparticles (LNPs), polymers, or biological vectors, is highlighted. Such advancements could improve delivery efficiency to a wider range of organs and increase the potential to treat more diseases. Early strategies for siRNA delivery focused primarily on LNPs, which have been widely accepted in clinical practice due to their low immunogenicity, increased circulation time, and in vivo stability. By optimizing the lipid-nucleic acid ratio, LNPs can be customized for targeted delivery, thereby enhancing therapeutic potential. The goal of LNP design is to address issues related to the delivery of the initial nucleic acid, such as instability caused by biological degradation mechanisms and difficulty in penetrating membrane barriers. Common types of LNPs include solid lipid nanoparticles, nanostructured lipid carriers, and cationic lipid-nucleic acid complexes, which are characterized by more complex internal structures and enhanced physical stability. However, these next-generation LNPs still face several challenges: 1) RNA size, LNP formulation, size, and polydispersity index significantly affect encapsulation efficiency; 2) Existing LNPs lack targeting ability and are not effectively delivered to specific cells; and 3) storage methods, the selection of cryoprotectants, and storage temperature can affect the long-term stability of the formulation. Therefore, it is necessary to develop genetic material delivery vehicles that solve the aforementioned problems while enhancing the ability to target specific organs and cells. Figure 1a shows the results of confirming the transdermal delivery effect of hyaluronic acid labeled with Rhodamine B (RhoB) into the dermal layer. FIG. 1b is a schematic diagram showing that the entry of a genetic material carrier into a cell according to the present invention occurs through endocytosis. FIG. 1c is a schematic diagram relating to a method for manufacturing a dielectric material carrier according to the present invention. Figure 2a shows the morphology of cationic solid lipid nanoparticles (CSLN) and genetic material carriers (HA/CSLN/siRNA) as seen by TEM (scale bar = 50 μm). Figure 2b shows the results of agarose gel electrophoresis at various weight ratios of CSLN/siRNA complexes (CSLN:siRNA = 1:3, 6, 9, 12, 15). Figure 2c shows the size distribution of CSLN, CSLN/siRNA complex, and HA/CSLN/siRNA, respectively. Figure 2d shows the zeta-potential (ζ-potential) measurement results for CSLN, CSLN/siRNA complex, and HA/CSLN/siRNA, respectively. Figure 2e shows the viability of B16F10 cells according to the concentrations of siRNA, CSLN/siRNA complex and HA/CSLN/siRNA. Figure 3a shows the results of fluorescence confocal microscopy analysis of siRNA labeled Cy3, CSLN/siRNA complex, HA/CSLN/siRNA (Example 1), and HA pretreatment (Example 2) (scale bar = 25 μm). Figure 3b shows the flow cytometry results of siRNA labeled Cy3, CSLN/siRNA complex, HA/CSLN/siRNA (Example 1), and HA pretreatment (Example 2). Figure 3c shows mRNA levels analyzed by RT-PCR in B16F10 cells after treatment with the CSLN/siVEGF c