Monkeypox is a disease caused by human infection with monkeypox virus. As an infectious disease (zoonotic infectious disease), monkeypox can be suffered by both humans and vertebrates (such as monkeys).1 Monkeypox virus is classified as Orthopoxvirus in Poxviridae. There are four viruses in Orthopoxvirus that cause disease to humans, and the other three are smallpox virus, vaccinia virus and cowpox virus2.

I7L protein is a Cysteine protease composed of 423 amino acids with a size of about 47 kDa. The phylogenetic tree analysis of Monkeypox I7L shows that it belongs to the same source as other I7Ls. At present, the gene product of I7L has become the core protease of cowpox virus3.

I7L is an essential late-stage gene, as shown by temperature-sensitive mutated viruses and conditional lethal mutated viruses. In the absence of I7L, the viral maturation process is blocked before forming a mature form. Proteases are recognized as ideal antiviral targets for antiviral therapy, because they cut the basic function of precursor multi-protein in virus replication, and protease inhibitors in other viruses (such as HIV) have shown the prospect of virus protease inhibitors for other viruses.3 Therefore, Monkeypox virus protease I7L is also recognized as an ideal target against monkeypox for inhibiting virus replication.

Monkeypox is currently spreading on a small scale in China, and we need to prevent and change it, and conduct research. Herein, A drug screening basing on small molecule drug reactions was performed to discover the drug can inhibit the virus.4 This project does not require direct contact with viruses, ensuring security and making research more secure. The spirit of science, together with new scientific knowledge and scientific thought, is a synthesis of values and norms contained in the deep structure and surface structure of scientific culture. We hope to make contributions to the cure of monkeypox, reduce people's suffering from illness, improve the cure rate of monkeypox, and curb the spread of the entire monkeypox virus, resulting in suppression of monkeypox virus transmission.

Based on the recent research, a small molecule compound TTP-6171 has been discovered, which exhibited selective inhibitory activity against orthopoxvirus, and TTP-6171 treatment (3-12 μM) can inhibit 50% of viral infections3. However, there is still a lack of data about its efficiency in vitro inhibitor screening models, and new screening models need to be designed to develop new antiviral molecules. Thus we decided to utilize Escherichia coli to develop a method for screening other proteins or potential compounds which might possess the ability to inhibit the monkeypox virus.

First, we optimized the monkeypox virus I7L protein sequence, constructed a cloning recombinant vector and transferred it into E. coli for expression. We isolated the I7L-His tag protein in vitro and finally verified its activity through the principle of fluorescence quenching. In the future, we will further use the obtained protein Develop screening kits for monkeypox virus inhibitors.

The implementation of this project contributes to promoting high-throughput drug screening targeting I7L, achieving the development and application of reagent kits, and drug discovery.

We will address the safety issues in the screening process of new drugs for monkeypox which eliminates the need for researchers to direct contact with the virus. It also provides a guarantee that our drugs inhibit monkeypox virus, and improves production and screening efficiency.

Nowadays, there is no drug that can completely cure patients with monkeypox. We are committed to helping society and alleviating patient pain.

1. Adnan N, Haq ZU, Malik A, et al. Human monkeypox virus: An updated review. Medicine (Baltimore). Sep 2 2022;101(35):e30406. doi:10.1097/MD.0000000000030406

2. Lum FM, Torres-Ruesta A, Tay MZ, et al. Monkeypox: disease epidemiology, host immunity and clinical interventions. Nat Rev Immunol. Oct 2022;22(10):597-613. doi:10.1038/s41577-022-00775-4

3. Katritch V, Byrd CM, Tseitin V, et al. Discovery of small molecule inhibitors of ubiquitin-like poxvirus proteinase I7L using homology modeling and covalent docking approaches. J Comput Aided Mol Des. Oct-Nov 2007;21(10-11):549-58. doi:10.1007/s10822-007-9138-7

4. Lam HYI, Guan JS, Mu Y. In Silico Repurposed Drugs against Monkeypox Virus. Molecules. Aug 18 2022;27(16)doi:10.3390/molecules27165277