KR-102963111-B1 - Pharmaceutical composition for treating inflammatory disease comprising polypeptide derived from Mycobacterium tuberculosis

Abstract

The present invention relates to a polypeptide derived from Mycobacterium tuberculosis or a variant thereof and a pharmaceutical composition for treating inflammatory diseases containing the same. The pharmaceutical composition according to the present invention can be applied as a therapeutic agent for inflammatory diseases by specifically binding to NLRP3 and inhibiting the activation of the NLRP3 inflammasome.

Inventors

• 김효근
• 양철수

Assignees

• 한양대학교 에리카산학협력단

Dates

Publication Date

20260512

Application Date

20230810

Priority Date

20230601

Claims (13)

1. An NLRP3-specific binding molecule comprising the sequence represented by SEQ ID NO. 1, which specifically binds to the NLRP3 (NOD-, LRR- and pyrin domain-containing protein 3) protein.
2. In claim 1, the binding molecule is an NLRP3-specific binding molecule composed of the sequence represented by SEQ ID NO. 1.
3. In claim 1, the binding molecule is an NLRP3-specific binding molecule comprising the sequence represented by SEQ ID NO. 2.
4. An NLRP3-specific binding molecule according to claim 1, wherein the amino acids at the 4th, 6th, and 7th positions of SEQ ID NO. 1 are not substituted.
5. An NLRP3-specific binding molecule according to claim 4 that does not include Q4L, V6H, or Q7L substitutions.
6. delete
7. A pharmaceutical composition for treating inflammatory diseases comprising an NLRP3-specific binding molecule according to any one of claims 1 to 5.
8. A pharmaceutical composition for treating an inflammatory disease according to claim 7, wherein the inflammatory disease is Alzheimer's disease, atherosclerosis, asthma and allergic airway inflammation, gout, inflammatory bowel disease, non-alcoholic fatty liver disease and non-alcoholic steatohepatitis, myocardial infarction, multiple sclerosis or experimental autoimmune encephalitis, nephropathy, hyperinflammation after influenza infection, graft-versus-host disease, stroke, silicosis, type 1 diabetes, obesity-induced inflammation or insulin resistance, rheumatoid arthritis, contact hypersensitivity, or traumatic brain injury.
9. A nucleic acid molecule expressing an NLRP3-specific binding molecule according to any one of claims 1 to 5.
10. A pharmaceutical composition for treating inflammatory diseases comprising the nucleic acid molecule of claim 9.
11. A pharmaceutical composition for treating an inflammatory disease according to claim 10, wherein the inflammatory disease is Alzheimer's disease, atherosclerosis, asthma and allergic airway inflammation, gout, inflammatory bowel disease, non-alcoholic fatty liver disease and non-alcoholic steatohepatitis, myocardial infarction, multiple sclerosis or experimental autoimmune encephalitis, nephropathy, hyperinflammation after influenza infection, graft-versus-host disease, stroke, silicosis, type 1 diabetes, obesity-induced inflammation or insulin resistance, rheumatoid arthritis, contact hypersensitivity, or traumatic brain injury.
12. A recombinant vector comprising the nucleic acid molecule of claim 9.
13. Isolated host cell comprising the nucleic acid molecule of claim 9.

Description

Pharmaceutical composition for treating inflammatory disease comprising polypeptide derived from Mycobacterium tuberculosis The present invention relates to a polypeptide derived from Mycobacterium tuberculosis that specifically binds to NLRP3, and a pharmaceutical composition for treating inflammatory diseases containing the same. Four types of inflammasomes are known: NLRP1, NLRP3, NLRP4, and Aim2, all of which contain PRRs belonging to the NLR family. Among these inflammasomes, the NLRP3 inflammasome plays a pivotal role in shaping immune responses and regulating the integrity of intestinal homeostasis in many common inflammatory diseases. NLRP3, a multiprotein complex composed of an NLRP3 scaffold, an adapter (ASC - apoptosis speck-like protein), and an effector (procaspase-1), interacts with the ASC to initiate the formation of the inflammasome. The ASC recruits and activates procaspase-1 to produce active caspase-1, which converts the cytokine precursors pro-IL-1β and pro-IL-18 into mature, biologically active IL-1β and IL-18, respectively. Once activated, active IL-1β and IL-18 trigger a series of inflammatory responses and pyogenic apoptosis. NLRP3 inflammasomes are produced by bone marrow-derived macrophages in the presence of stimulation by microbial and non-microbial factors, such as bacterial toxins, particulate matter, and lipopolysaccharides (LPS). Although the mechanism of NLRP3 activation has not yet been clearly elucidated, several molecular and cellular events have been proposed to explain their involvement in inflammasome activation, including K + efflux, Ca2 + signaling, mitochondrial dysfunction, and reactive oxygen species (ROS) production. For example, particulate matter induces endocytosis and damage to the lysosomal membrane, activating NLRP3 inflammasomes to release cathepsin B into the cytoplasm. Figure 1A shows the results of confirming the binding of NLRP3 and Rv0747/PE_PGRS10, Figure 1B shows the results of confirming the binding site with Rv0747/PE_PGRS10 through a deletion mutation of NLRP3, Figure 1C shows the results of confirming the binding activity of NLRP3 according to the position of Rv0747/PE_PGRS10, and Figure 1D shows the peptide sequence predicted as the site in Rv0747/PE_PGRS10 that exhibits binding ability with NLRP3. FIG. 2A is the result of confirming the degree of binding between NLRP3 and ASC following peptide treatment according to one embodiment of the present invention, FIG. 2B and FIG. 2C are the result of confirming the specific secretion inhibitory ability against IL-1β, IL-18, and caspase-1 following peptide treatment according to one embodiment of the present invention, and FIG. 2D is the result of confirming that the peptide according to one embodiment of the present invention has superior binding ability to NLRP3 compared to ASC. Figure 3 shows the results of confirming the change in ubiquitination (Figure 3A) and phosphorylation (Figure 3B) of the NRR domain by peptide treatment according to one embodiment of the present invention. Figure 4 is the result of identifying the amino acid positions essential for binding with NLRP3 among the peptides according to one embodiment of the present invention. FIGS. 5 and 6 show peptide and protein sequences and nucleic acid sequences encoding them according to one embodiment of the present invention. Hereinafter, the present invention will be described in detail with reference to the attached drawings for embodiments of the present invention. However, the following embodiments are presented as examples of the present invention, and if it is determined that a detailed description of a technology or configuration well known to those skilled in the art may unnecessarily obscure the essence of the present invention, such detailed description may be omitted, and the present invention is not limited thereby. The present invention is capable of various modifications and applications within the scope of the claims set forth below and the equivalents interpreted therefrom. Furthermore, the terminology used in this specification is used to appropriately describe preferred embodiments of the present invention, and may vary depending on the intent of the user or operator, or the conventions of the field to which the present invention belongs. Accordingly, the definitions of these terms should be based on the content throughout this specification. Throughout the specification, when a part is described as "comprising" a certain component, unless specifically stated otherwise, this means that it does not exclude other components but may include additional components. All technical terms used in this invention, unless otherwise defined, are used in the sense generally understood by those skilled in the art in the relevant field of this invention. Additionally, while preferred methods or samples are described herein, similar or equivalents are also included within the scope of this invention. The contents of all publications c