Unlock instant, AI-driven research and patent intelligence for your innovation.

Application of histone deacetylase-4 inhibitor in the preparation of medicaments for the treatment of multiple myeloma

A multiple myeloma, deacetylase technology, applied in the field of biomedicine, can solve problems such as increased autophagy, apoptosis, and increased energy requirements of the body

Active Publication Date: 2019-09-27
SECOND AFFILIATED HOSPITAL SECOND MILITARY MEDICAL UNIV
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In multiple myeloma, the malignant proliferation of plasma cells produces a large number of immunoglobulins, causing endoplasmic reticulum stress and increased misfolded proteins. Cells need autophagy to clear these proteins to maintain survival. In addition, the rapid proliferation of myeloma cells and A large amount of immunoglobulin is synthesized, and the energy demand of the body increases, which also increases autophagy. At this time, autophagy can protect cells, but when the external stimulus is too strong, excessive autophagy may lead to apoptosis. When the folded protein exceeds the processing capacity of proteasome and autophagy, it will cause the accumulation of misfolded protein and excessive autophagy, which makes autophagy switch from inhibiting apoptosis to inducing death (Grandér D, Kharaziha P, Laane E, et al. as the main means of cytotoxicity by glucocorticoids in hematological malignancies. Autophagy. 2009 Nov; 5(8):1198-200.)

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of histone deacetylase-4 inhibitor in the preparation of medicaments for the treatment of multiple myeloma
  • Application of histone deacetylase-4 inhibitor in the preparation of medicaments for the treatment of multiple myeloma
  • Application of histone deacetylase-4 inhibitor in the preparation of medicaments for the treatment of multiple myeloma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1. Search and verification of miR-145-3p target genes

[0047] Bioinformatics method to predict the target genes of microRNAs: use TargetScan, Bibiserve, Pictar and other microRNA target gene prediction websites and software to predict and analyze the secondary structure and target genes of microRNAs, and search for high sequence matching, stable secondary structures, and target genes. Genes whose sequences are highly conserved across species were subsequently verified.

[0048] The target gene of microRNA is detected by double fluorescent reporter gene method: the sequence that can interact with microRNA in the 3' untranslated region (Untranslated Region, UTR) of the target gene is cloned into the 3'UTR of luciferase in the pGL3 plasmid, and the recombinant Luciferase reporter gene plasmid. Co-transfect 293T cells with the pSuper vector expressing the corresponding microRNA in a certain ratio. After 24 hours, the cells were lysed, and the expression level of ...

Embodiment 2

[0056] Example 2: Design, synthesis and inhibition efficiency verification of siRNA

[0057] The full sequence and mRNA sequence of HDAC4 were retrieved from NCBI GeneBank, the biological analysis of HDAC4 was carried out using existing network resources and commonly used software, and the coding region was selected as the target sequence for siRNA design. Refer to the siRNA design principle, and compare it with the human genome sequence through the blast function of the GeneBank database to ensure that there is no homology; exclude potential siRNAs that have 8 consecutive bases at the 5' end of the aitisense chain paired with other genes; exclude any consecutive 14 bases base-pairing potential siRNAs with other genes.

[0058] The siRNA sequence designed and synthesized in this example is as follows, and was commissioned to be synthesized by Guangzhou Ruibo Biotechnology Co., Ltd.:

[0059]

[0060] The inhibition efficiency of the three siRNAs was verified by the followi...

Embodiment 3

[0069]Example 3: Regulation of MM cell proliferation ability after HDAC4 interference

[0070] By transfecting HDAC4-siRNA, observe the regulation of cell proliferation by interfering with HDAC4. The implementation process is as follows:

[0071] (1) LP-1 cells in the logarithmic growth phase were collected and seeded in a 96-well plate at a density of 20,000 per well.

[0072] (2) Transfection was mediated by RNAimax transfection reagent, and cell proliferation was detected at 72 hours.

[0073] (3) 110 μL of fresh medium containing 10 μL of CCK-8 was added to each well, and after culturing for 3 hours, the absorbance value of each well was detected at a wavelength of 560 nm with a multifunctional microplate reader.

[0074] (4) The experiment was repeated 3 times,

[0075] The experimental results showed that, compared with the negative control group, the absorbance value of the cells transfected with HDAC4-siRNA group was significantly reduced. The negative control group ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the technical field of biological medicine, and in particular relates to application of a histone deacetylase-4 (HDAC4) inhibitor in the preparation of a medicament for treating multiple myeloma (MM) disease. According to the application, firstly, the HDAC4 is found to possibly be a target spot for treating the MM; secondly, aiming at an HDAC4 gene, siRNA (Small interfering Ribonucleic Acid) for inhibiting the expression of the HDAC4 gene is designed; the siRNA can be used for inhibiting the proliferation of multiple myeloma cells, inducing cell apoptosis to generate autophagy, and causing the up-regulation of downstream transcription factors. The invention provides a new target spot for preventing and treating the MM disease and also provides a stable, effective and low-toxicity biological targeting preparation or medicament for inhibiting the expressing of the HDAC4 and treating the MM.

Description

technical field [0001] The invention relates to the technical field of biomedicine, and more specifically relates to the application of histone deacetylase-4 inhibitors in the preparation of drugs for treating multiple myeloma diseases. Background technique [0002] Multiple myeloma (Multiple myeloma, MM) is a malignant blood disease characterized by abnormal clonal proliferation of plasma cells in the bone marrow (BM), secreting a large amount of monoclonal immunoglobulin or its fragments, causing kidney, bone, etc. Damage to tissues or organs. The incidence of MM accounts for about 10% of all hematological malignancies, and it is more likely to occur in the elderly. With the progress of population aging in my country, the number of MM incidences tends to increase. The common clinical manifestations are bone pain, anemia, renal insufficiency or low immunity. At present, the main means of treating MM are hormones, immunomodulators, alkyl drugs, protease inhibitors and so o...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61K48/00A61K31/7088A61P35/00C12N15/113
Inventor 周琳吴洪坤刘畅樊笑霞吴雯吴萍
Owner SECOND AFFILIATED HOSPITAL SECOND MILITARY MEDICAL UNIV