KR-20260065570-A - Nano-graphene oxide-based composition for treating post-transplant complications and uses thereof

Abstract

The present invention relates to nanographene oxide having a therapeutic effect on post-transplant complications such as graft-versus-host disease, nanographene oxide with a surface modified with a zwitter ionic polymer, macrophages treated with said nanographene oxide, and applications thereof. The present invention confirmed that the administration of nanographene oxide (NGO), an NGO modified with a zwitter ionic polymer, or said NGO-treated macrophages in an animal model of acute graft-versus-host disease (GVHD) occurring after transplantation improved survival rates and effectively reduced tissue damage and inflammation. The NGO regulated the activation and differentiation direction of immune cells, particularly macrophages, thereby reducing inflammation-inducing (M1-type) macrophages and increasing the induction of immunomodulatory (M2-type) macrophages and regulatory T cells (Treg), and also provided an effect of restoring immune balance in patient-derived immune cells. Therefore, the present invention can be effectively utilized for the treatment of immune diseases caused by post-transplant complications such as graft-versus-host disease.

Inventors

• 유경록
• 유아론
• 유재철
• 최순원

Assignees

• 서울대학교산학협력단
• 인비씨티 주식회사

Dates

Publication Date

20260508

Application Date

20251030

Priority Date

20241030

Claims (15)

1. A pharmaceutical composition for preventing or treating post-transplant complications comprising nanographene oxide.
2. A composition according to claim 1, characterized in that the average diameter of the nano-graphene oxide is 1 to 100 nm and the thickness is 0.3 to 3 nm.
3. A composition according to claim 1, wherein the post-transplant complication is one or more selected from the group consisting of graft-versus-host disease, graft rejection, autoimmune disease, posttransplant lymphoproliferative disorder, anemia, hyperglycemia, veno-occlusive disease, and pulmonary fibrosis.
4. A pharmaceutical composition for preventing or treating post-implantation complications comprising nano-graphene oxide with a surface modified by a zwitter ionic polymer.
5. A composition according to claim 4, wherein the nano-graphene oxide modified with the amphoteric ionic polymer is a nano-graphene oxide variant in which nano-graphene oxide and the amphoteric ionic polymer are combined in a ratio of 1.0:0.01 to 1.0:100.0.
6. A composition according to claim 4, wherein the amphoteric ionic polymer is one or more selected from the group consisting of PMPC (Poly(2-methacryloyloxyethyl phosphorylcholine)), PVA (Polyvinyl alcohol), PAMPS (Poly(acrylamidomethylpropane sulfonic acid)), PDMAEMA (Poly(N,N-dimethylaminoethyl methacrylate)), PEGMA (Poly(ethylene glycol) methyl ether methacrylate), PHEMA (Poly(2-hydroxyethyl methacrylate)), and PEI (Poly(ethylene imine)).
7. A pharmaceutical composition for the prevention or treatment of post-transplant complications comprising macrophages treated with nanographene oxide.
8. A composition according to claim 7, characterized in that the average diameter of the nano-graphene oxide is 1 to 100 nm and the thickness is 0.3 to 3 nm.
9. A composition according to claim 7, wherein the surface of the nano-graphene oxide is modified with a zwitter ionic polymer.
10. A cell therapy agent for treating post-transplant complications comprising macrophages treated with nanographene oxide.
11. i) a step of producing immature macrophages by culturing monocytes with growth factors for 4 to 8 days; and ii) a step of treating the above immature macrophages with 15–25 μg/mL nanographene oxide for 16–20 hours; A method for preparing macrophages treated with nanographene oxide for the prevention or treatment of post-transplant complications, comprising
12. A method of preparation according to claim 11, characterized in that the monocytes are obtained from one or more selected from the group consisting of peripheral blood, spleen, and bone marrow.
13. A method of preparation according to claim 11, characterized in that the growth factor is a macrophage-colony stimulating factor.
14. A manufacturing method according to the 11th, characterized in that the average diameter of the nano-graphene oxide is 1 to 100 nm and the thickness is 0.3 to 3 nm.
15. A manufacturing method according to claim 11, characterized in that the surface of the nano-graphene oxide is modified with a zwitter ionic polymer.

Description

Nano-graphene oxide-based composition for treating post-transplant complications and uses thereof The present invention relates to nano-graphene oxide having a therapeutic effect on post-transplant complications such as graft-versus-host disease, nano-graphene oxide with a surface modified with a zwitter ionic polymer, macrophages treated with said nano-graphene oxide, and applications thereof. The present invention was completed with the support of the research project No. 500-20230269 (500-20240247) and the project title “Development of a side-effect-free cell therapy for graft-versus-host disease and a coronavirus treatment using nanographene oxide (Elucidation of the inflammation-regulating mechanism of nanographene oxide for graft-versus-host disease and development of a treatment for porcine epidemic diarrhea virus)” and the support of INBCT. Allogeneic hematopoietic stem cell transplantation (HSCT) is a widely used treatment for various malignant and non-malignant hematological and immune diseases. However, the infusion of donor-derived immune cells carries significant risks as they may recognize the recipient's tissue as a foreign substance. This reaction is primarily caused by human leukocyte antigen mismatches and minority histocompatibility antigen (HSA) differences, ultimately leading to graft-versus-host disease (GVHD). Despite advancements in immunosuppressive therapy for GVHD prevention, acute GVHD (aGVHD) still occurs in 40–50% of transplant patients and remains a major cause of death after allogeneic HSCT, excluding recurrence. The standard first-line treatment for severe acute GVHD is systemic steroid therapy, but it does not respond in 35–60% of patients. Although ruxolitinib has been approved as a treatment for steroid-refractory GVHD, the disease still has a high mortality rate, and a universally effective second-line treatment has not been established. The pathogenesis of acute GVHD consists of a three-step chain process. First, tissue damage caused by pretreatment, such as whole-body radiation or chemotherapy, activates the recipient's antigen-presenting cells (APCs). Subsequently, donor-derived alloreactive T cells are stimulated to initiate the afferent phase of the immune response. Finally, target organs are damaged by the secretion of inflammatory cytokines and the release of cytotoxic granules. Activated donor T cells differentiate into Th1 and Th17 helper T cell subtypes, causing host tissue damage. In contrast, regulatory T cells (Tregs) perform immunomodulatory functions that suppress GVHD through contact-dependent and contact-independent mechanisms. In addition to T cells, macrophages are emerging as important regulators in the pathogenesis of aGVHD. Macrophages possess remarkable heterogeneity and plasticity, playing a crucial role in both innate and adaptive immunity. They are classified into inflammatory M1 and anti-inflammatory M2 subtypes based on their activation state. M1 macrophages are activated by IFN-γ and TLR ligands and secrete inflammatory cytokines such as TNF-α and IL-12 through the IFN-γ/STAT1 and LPS/TLR4/MyD88 signaling pathways. M1 macrophage infiltration serves as a biomarker of disease progression, and it has been reported that higher M1/M2 ratios are associated with increased severity (grades II-IV) of aGVHD. Conversely, mesenchymal stem cell-derived extracellular vesicles (EVs) induce macrophage polarization toward M2, thereby mitigating aGVHD, while donor-derived M2 macrophages have also been shown to suppress GVHD in mouse models. These results suggest that macrophages are potential therapeutic targets, but it has not been clearly revealed how the IFN-γ/STAT1 axis is involved in M1 macrophage-mediated inflammation. Against this backdrop, therapeutic approaches involving the direct injection of M2 macrophages or the repolarization of M1 macrophages into M2 macrophages within the body are being attempted. However, macrophage-based therapies face an inherent limitation due to the plasticity of macrophages, which change their phenotypes depending on the surrounding environment. Even M2 macrophages administered for therapeutic purposes can revert to an inflammatory phenotype similar to M1 macrophages if exposed to a strong inflammatory microenvironment (e.g., IFN-γ, DAMPs, etc.). This phenotypic instability can nullify therapeutic effects or even exacerbate inflammation, leaving the technology to maintain a stable anti-inflammatory phenotype as a key challenge in the development of macrophage therapies. Nano-sized graphene oxide (NGO) is a carbon-based nanoparticle that is attracting attention as a potential therapeutic candidate for various inflammatory diseases, such as inflammatory bowel disease and liver injury, due to its excellent biocompatibility and context-dependent immune-modulating properties. NGOs possess superior reactive oxygen species (ROS) scavenging capabilities resulting from their electron-deficient ring structure, thereby alleviating o