Chemical small molecule composition and method for directly reprogramming in-vitro and in-vivo chemically induced fibroblasts into hepatocytes

A fibroblast, chemically induced technology, applied in biochemical equipment and methods, animal cells, drug combinations, etc., can solve the problems of incomplete differentiation, high cost and low differentiation efficiency of stem cells

Pending Publication Date: 2021-03-12
TRANSCEND CYTOTHERAPY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing directed differentiation methods have one or more of the following defects: such as low differentiation efficiency, low transformation purity, lack of function of differentiated hepatocytes, possible immune rejection, and potential cancer risk due to incomplete differentiation of stem cells And the high cost, etc., cannot meet the clinical needs
Fibrotic diseases caused by abnormal fibroblasts or fibroblasts, such as liver cirrhosis (hepatic fibrosis), currently lack effective therapeutic drugs and means in clinical practice, and it is urgent to develop effective new methods and new drugs

Method used

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  • Chemical small molecule composition and method for directly reprogramming in-vitro and in-vivo chemically induced fibroblasts into hepatocytes
  • Chemical small molecule composition and method for directly reprogramming in-vitro and in-vivo chemically induced fibroblasts into hepatocytes
  • Chemical small molecule composition and method for directly reprogramming in-vitro and in-vivo chemically induced fibroblasts into hepatocytes

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Experimental program
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preparation example Construction

[0126] The preparation method of the small molecule composition of the present invention is determined according to the dosage form to be prepared and the route of administration. Those skilled in the art, after referring to the combination and proportioning provided by the present invention, adopt the conventional preparation method of the pharmaceutical composition The chemical small molecule composition of the present invention can be prepared.

[0127] It should be understood that although in the specific embodiment, the inventor listed several composition forms, those skilled in the art can also deduce from this that any other small molecule combination form of the present invention also has outstanding effects.

[0128] The present inventors have confirmed for the first time that the small molecule composition of the present invention can induce fibroblasts to transform into hepatocytes in situ in vivo, and reduce or reduce the fibroblasts or fibroblasts in tissues / organs...

Embodiment 1

[0156] Example 1. Small molecule composition for direct reprogramming (transdifferentiation) of fibroblasts induced by small molecules into hepatocytes, and preparation of transdifferentiation medium and reagents thereof

[0157] Design the following composition or transdifferentiation medium, which can be prepared according to molar concentration or weight concentration:

[0158] 1. Formula of small molecule composition for fibroblast transdifferentiation liver cell

[0159] (1) Fibroblast transdifferentiation liver cell composition 1

[0160] GSK3β inhibitor CHIR-99021: final concentration 3 μM;

[0161] G9aHMT inhibitor BIX01294: final concentration 3 μM;

[0162] (2) Fibroblast transdifferentiation liver cell composition 2

[0163] GSK3β inhibitor LY2090314: final concentration 0.3 μM;

[0164] G9aHMT inhibitor UNC0642: final concentration 2 μM;

[0165] (3) Fibroblast transdifferentiation liver cell composition 3

[0166] GSK3β inhibitor LiCl: final concentration 10...

Embodiment 2

[0212] Example 2, Fibroblast Transdifferentiation Hepatocyte Medium 1, 11 Chemically induced hepatic fibroblast (hepatic stellate cell HSC) transdifferentiation into hepatocyte comparative experiment

[0213] 1. Fibroblast transdifferentiation hepatocyte culture

[0214] Mouse hepatic stellate cells (HSCs), that is, hepatic fibroblasts, were suspended in DMEM, a cell base medium containing 10% calf serum, plated, and cultured at 37°C. Set up control group and treatment group;

[0215] After hepatic stellate cells adhered to the wall, the original medium was discarded, and the treatment groups were replaced with Fibroblast Transdifferentiated Hepatocyte Culture Substrates 1 and 11 were cultured at 37°C, and the medium was changed every 3 days; passage was performed once every 3-7 days.

[0216]2. Subculture of Hepatic Stellate Cells (HSC) Transdifferentiated Hepatocytes

[0217] Subculture steps: Discard the original culture medium, wash once with PBS, add cell digestion so...

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Abstract

The invention relates to a chemical small molecule composition and a method for chemically inducing in-vitro and in-vivo fibroblasts to be directly reprogrammed (transdifferentiated) into hepatocytesonly by using chemical small molecules (small molecules for short) without introducing any exogenous gene / transcription factor / MicroRNA gene, RNA, protein, polypeptide and other inducible factors. Themicromolecule composition induces fibroblasts to be transdifferentiated into hepatocytes in vitro or chemically induces in-situ fibroblasts to be transdifferentiated into hepatocytes in vivo, fibrosis of fibrotic tissues or organs is reduced or reduced through the in-situ transdifferentiation effect of the hepatocytes in vivo, and the effect of treating fibrotic diseases such as hepatic fibrosis / liver cirrhosis is achieved. The small molecule composition can be added with a drug carrier and an excipient to prepare a drug or a prodrug for treating fibrotic diseases, or added with normal saline, a buffer solution, a cell basal culture medium and the like to prepare a transdifferentiation reagent or a culture medium; the obtained transdifferentiated hepatocytes can provide hepatocyte sourcesfor scientific research, medicine, clinical hepatocyte transplantation and the like.

Description

technical field [0001] The present invention belongs to the interdisciplinary fields of cell biology, stem cell biology (cell reprogramming), medicine, and pharmacy; more specifically, the present invention relates to a method that does not introduce or use any exogenous gene / transcription factor / MiRNA (miRNA) gene, Or its gene combined with a carrier, or its RNA, protein, polypeptide and other induction factors, does not use any cytokines or growth factors, only chemical small molecules (small molecules for short) combination, chemically induce fibroblasts in vivo and in vitro to directly regenerate Chemical small molecule compositions and methods for programming (transdifferentiation) into hepatocytes. Background technique [0002] China is a country with a large number of liver diseases. There are a large number of patients with hepatitis A, hepatitis B, fatty liver, alcoholic liver, liver cirrhosis (liver fibrosis), and acute and chronic liver failure caused by various l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/067C12N2501/727C12N2501/72C12N2506/1307C12N2506/13A61K35/545A61P1/16
Inventor 张培霖
Owner TRANSCEND CYTOTHERAPY CO LTD
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