CN115029299A - Application of JAK2 inhibitor in induced differentiation of islet beta cells - Google Patents

Application of JAK2 inhibitor in induced differentiation of islet beta cells Download PDF

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CN115029299A
CN115029299A CN202210812902.XA CN202210812902A CN115029299A CN 115029299 A CN115029299 A CN 115029299A CN 202210812902 A CN202210812902 A CN 202210812902A CN 115029299 A CN115029299 A CN 115029299A
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islet beta
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jak2
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beta cells
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CN115029299B (en
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杜媛媛
徐栋
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Hangzhou Ruipu Chenchuang Technology Co ltd
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Abstract

The invention relates to the technical field of biology, in particular to application of a JAK2 inhibitor in pancreatic islet beta cell induced differentiation. The JAK2 inhibitor in the invention comprises at least one selected from CEP-33779, TG101209 and TG 101348. The JAK2 inhibitor can greatly improve the differentiation efficiency of endocrine cells to islet beta cells, so that the differentiation of the islet beta cells in vivo can be regulated and controlled, and the method is applied to the field of disease treatment.

Description

Application of JAK2 inhibitor in induced differentiation of islet beta cells
Technical Field
The invention relates to the technical field of biology, in particular to application of a JAK2 inhibitor in inducing differentiation of islet beta cells.
Background
Diabetes is one of the major diseases affecting human health, and insulin hyposecretion and glucose metabolism disorder caused by the decrease of pancreatic endocrine beta cell function are important causes of diabetes, and it is an important part of the research on the pathogenesis and the functional state of pancreatic islet cells. Among them, the treatment of diabetes using islet beta cells is an effective method.
However, the differentiation to obtain islet beta cells requires further investigation.
Disclosure of Invention
The present invention is directed to solving, at least to some extent, one of the technical problems in the related art. Therefore, the invention aims to provide the application of the JAK2 inhibitor in the field of islet beta cell induced differentiation and the application of the JAK2 inhibitor in preparing a medicament for preventing and/or treating diabetes.
The inventor of the invention finds out in the research process that: the JAK2 inhibitor can play an important role in the process of directional differentiation of Pancreatic precursor cells (Pancreatic progeniors) to islet beta cells (Pancreatic beta cells), and the JAK2 inhibitor can greatly improve the efficiency of differentiation of endocrine cells to the islet beta cells, so that cells to be acted express specific molecular markers of Proinsulin (Proinsulin), Insulin (Insulin), C-Peptide (C-Peptide) and NKX 6.1. Therefore, the JAK2 inhibitor can regulate the directional differentiation of the islet beta cells in vivo or in vitro, and further can be used for preventing or treating diseases caused by the deletion or abnormality of the islet beta cells.
To this end, according to a first aspect of the invention, the invention provides the use of a JAK2 inhibitor in the field of induced differentiation of islet beta cells. The inventor finds that the JAK2 inhibitor can greatly improve the differentiation efficiency of endocrine cells to islet beta cells, so that the differentiation of the islet beta cells in vivo can be regulated; or can be used to induce differentiation into islet beta cells in vitro for specific assays.
According to the embodiment of the invention, the application of the JAK2 inhibitor in the field of islet beta cell induced differentiation can be further added with the following technical characteristics:
in some embodiments of the invention, the JAK2 inhibitor is a JAK2 protein kinase inhibitor.
In some embodiments of the invention, the JAK2 protein kinase inhibitor comprises at least one selected from CEP-33779, TG101209, TG 101348. Representative substances of the JAK2 inhibitor, CEP-33779, TG101209, or TG101348, can inhibit JAK2 protein kinase, so that endocrine cells or pancreatic precursor cells can be induced to differentiate into islet beta cells, and the islet beta cells can be used as an induction differentiation agent or a regulatory factor to mediate the differentiation of the islet beta cells.
In some embodiments of the invention, the JAK2 inhibitor can be used to promote the directed differentiation of pancreatic endocrine precursor cells (pancreatic endocrine precursors) into islet beta cells. The term "pancreatic endocrine precursor cell" as used herein refers to a precursor cell having the potential to differentiate to obtain islet cells, which express a specific molecular marker for pancreatic endocrine.
In some embodiments of the invention, the JAK2 inhibitor is capable of promoting the committed differentiation of pancreatic precursor cells into islet beta cells.
According to a second aspect of the invention, the invention provides a use of a JAK2 inhibitor in the preparation of a medicament for the prevention and/or treatment of diabetes. The JAK2 inhibitor can greatly improve the differentiation efficiency of pancreatic endocrine precursor cells to islet beta cells, so that the differentiation of the islet beta cells in vivo can be regulated, and the JAK2 inhibitor can be used for treating diseases related to islet beta cell abnormality, such as diabetes.
According to the embodiment of the invention, the application of the JAK2 inhibitor in preparing the medicament for preventing and/or treating diabetes can be further added with the following technical characteristics:
in some embodiments of the invention, the JAK2 inhibitor is a JAK2 protein kinase inhibitor.
In some embodiments of the invention, the JAK2 protein kinase inhibitor comprises at least one selected from CEP-33779, TG101209, TG 101348.
In some embodiments of the invention, the JAK2 inhibitor can be used to promote the directed differentiation of pancreatic endocrine precursor cells into islet beta cells.
In some embodiments of the invention, the JAK2 inhibitor is capable of promoting the committed differentiation of pancreatic precursor cells into islet beta cells.
In a third aspect of the invention, the invention provides a pharmaceutical composition capable of inducing differentiation of cells into islet beta cells, comprising a JAK2 protein kinase inhibitor.
In some embodiments of the invention, the JAK2 protein kinase inhibitor comprises at least one of CEP-33779, TG101209, TG 101348.
In some embodiments of the invention, the above-described islet beta cell-inducing differentiation further comprises: other pharmaceutically acceptable carriers and/or excipients.
In a fourth aspect of the invention, the invention provides a kit which can be used to promote differentiation of cells into insulin beta cells, the kit comprising a JAK2 inhibitor.
According to an embodiment of the invention, the cells are selected from pancreatic endocrine precursor cells or pancreatic precursor cells.
According to an embodiment of the invention, in the kit, the JAK2 inhibitor comprises at least one selected from CEP-33779, TG101209, TG 101348.
In a fifth aspect of the present invention, the present invention provides a method of promoting differentiation of cells into islet beta cells, comprising: contacting and culturing the cells with a JAK2 inhibitor, such that islet beta cells are obtained.
In some embodiments of the present invention, the method for promoting differentiation of cells into islet beta cells described above may further comprise the following technical features:
in some embodiments of the invention, the cells are pancreatic endocrine precursor cells or pancreatic precursor cells.
In some embodiments of the invention, the cell is cultured in contact with the JAK2 inhibitor for 5 to 7 days. The cells and JAK2 inhibitor are continuously cultured for 5-7 days, so that pancreatic endocrine precursor cells or pancreatic precursor cells can be differentiated into islet beta cells, and the expression of the islet beta cells in vivo can be regulated. It can also be used to induce differentiation of cells in vitro, thereby acting as an inducer.
Drawings
FIG. 1 is a flow cytogram of increasing the expression level of the specific molecular markers Proinulin/Insulin/C-Peptide and NKX6.1 of islet beta cells by adding a chemical small molecule inhibitor related to the JAK2 signal path according to an embodiment of the present invention.
Figure 2 is a graph showing a significant analysis of the production of islet beta cells by the addition of a chemical small molecule inhibitor associated with the JAK2 signaling pathway, provided in accordance with an embodiment of the present invention.
FIG. 3 is a graph of islet beta cells obtained by continuous addition of TG101348 for 6 days, according to an embodiment of the present invention.
FIG. 4 is a graph showing that the addition of TG101348 increases the production ratio of islet beta cells, provided in accordance with an embodiment of the present invention.
FIG. 5 shows the content ratio of each molecular marker after TG101348 is added, wherein 5a shows the content ratio of NKX6.1 and proinsulin, insulin and C-peptide after TG101348 is added, and 5b shows the content ratio of NKX6.1 and C-peptide after TG101348 is added.
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the drawings are illustrative and intended to be illustrative of the invention and are not to be construed as limiting the invention.
The invention finds that the JAK2 inhibitor plays an important role in the process of directional differentiation of Pancreatic precursor cells (Pancreatic progenitors) to islet beta cells (Pancreatic beta cells), thereby providing the application of the JAK2 inhibitor in the field of induced differentiation of the islet beta cells, greatly improving the efficiency of differentiation of the Pancreatic endocrine precursor cells to the islet beta cells, and expressing specific molecular markers Proinsulin (Proinsulin), Insulin (Insulin), C-Peptide (C-Peptide) and NKX6.1 of the islet beta cells.
Therefore, the invention provides an application of the JAK2 inhibitor in preparing a medicament for preventing and/or treating diabetes. According to a specific embodiment of the invention, the invention provides the application of the JAK2 protein kinase inhibitor in the field of preparation of medicines for preventing and/or treating diabetes. According to embodiments of the invention, the JAK2 protein kinase inhibitor may be CEP-33779, TG101209, or TG 101348. JAK2 inhibitors, especially JAK2 protein kinase inhibitors, can induce pancreatic precursor cells or pancreatic endocrine precursor cells to differentiate into islet beta cells, thereby allowing the regulation of blood glucose balance for the prevention or treatment of diabetes.
Herein, the term "JAK 2 inhibitor" means all inhibitor substances capable of acting on the JAK2 signaling pathway, which substances are capable of inhibiting the expression of a signaling molecule in the JAK2 signaling pathway.
Herein, the term "JAK 2 protein kinase inhibitor" means that JAK2 protein kinase is targeted and is capable of inhibiting the activity of JAK2 protein kinase. According to an embodiment of the invention, the JAK2 protein kinase inhibitor comprises at least one selected from CEP-33779, TG101209, TG101348 as shown in table 1. According to an embodiment of the invention, the JAK2 protein kinase inhibitor is at least two selected from CEP-33779, TG101209, TG 101348.
Table 1 structure and related information for chemical small molecule inhibitors related to the JAK2 signaling pathway
Figure BDA0003730619140000051
According to a specific embodiment of the present invention, there is provided a pharmaceutical composition comprising a JAK2 protein kinase inhibitor, said pharmaceutical composition being capable of inducing differentiation of cells into islet beta cells.
According to an embodiment of the present invention, the pharmaceutical composition further comprises other pharmaceutically acceptable carriers and/or excipients. Different excipients are used for matching JAK2 protein kinase inhibitors, so that different pharmaceutical compositions can be prepared, and the pharmaceutical compositions can be prepared into different dosage forms according to the target administration route. Examples of routes of administration include, but are not limited to: parenteral routes, such as intravenous, intradermal, subcutaneous, intramuscular, subcutaneous, oral, buccal, sublingual, inhalation, intranasal, transdermal, topical, transmucosal, intratumoral, intrasynovial and rectal administration. In particular embodiments, the compositions are formulated in accordance with conventional procedures as pharmaceutical compositions suitable for intravenous, subcutaneous, intramuscular, oral, intranasal or topical administration to humans. In one embodiment, the pharmaceutical composition is formulated for subcutaneous administration to a human according to conventional methods. Typically, compositions for intravenous administration are solutions formulated with sterile buffers. If desired, the pharmaceutical composition may also contain a solubilizing agent and a local anesthetic to relieve pain at the site of injection.
The invention also provides a pharmaceutical dosage form. Examples of such pharmaceutical dosage forms include, but are not limited to: a tablet; a caplet; capsules, such as soft elastic gelatin capsules; a cachet; a lozenge; a dispersant; suppositories; an ointment; ointment; powder preparation; a dressing agent; a cream; a plaster; a solution; a patch; aerosols (e.g., nasal sprays or inhalants); gelling; liquid dosage forms suitable for oral or mucosal administration to a subject, including suspensions (e.g., aqueous or non-aqueous liquid suspensions, oil-in-water emulsions, or water-in-oil liquid emulsions); a liquid dosage form suitable for parenteral administration to a subject; and solid dosage forms.
In the case of oral dosage forms, pharmaceutical compositions of the invention suitable for oral administration are presented in discrete dosage forms, including, but not limited to: tablets (e.g., chewable tablets), caplets, capsules, and liquids (e.g., flavored syrups). In certain embodiments, the oral dosage form is a solid and is prepared with anhydrous ingredients under anhydrous conditions. The oral dosage forms of the invention are typically prepared by intimately mixing the active ingredient with at least one excipient according to conventional pharmaceutical compounding techniques. Excipients may take a wide variety of forms depending on the form of preparation desired for administration. For example, excipients suitable for oral liquid or aerosol dosage forms include, but are not limited to: water, glycols, oils, alcohols, flavoring agents, preservatives, and coloring agents. Examples of excipients suitable for use in solid oral dosage forms (e.g., powders, tablets, capsules, and caplets) include, but are not limited to: starch, sucrose, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, and disintegrating agents.
Examples of excipients that may be used in the oral dosage forms of the present invention include, but are not limited to: binder, filler, disintegrant and lubricant. Suitable binders for the pharmaceutical compositions and dosage forms include, but are not limited to: corn starch, potato starch or other starches, gelatin, natural and synthetic gums such as acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethylcellulose calcium, carboxymethylcellulose sodium), polyvinylpyrrolidone, methylcellulose, pregelatinized starch, hydroxypropylmethylcellulose, microcrystalline cellulose, and mixtures thereof.
Examples of fillers suitable for use in the pharmaceutical compositions and dosage forms described herein include, but are not limited to: talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextran, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures thereof. Disintegrants are used in the compositions of the invention to disintegrate the tablet when it is exposed to an aqueous environment. Tablets containing too much disintegrant disintegrate upon storage, while tablets containing too little disintegrant do not disintegrate at a desired rate or under desired conditions. Accordingly, an amount of disintegrant that is neither too much nor too little to adversely affect the release of the active ingredient should be used to form the solid oral dosage form of the present invention. Disintegrants that can be used in pharmaceutical compositions and dosage forms of the invention include, but are not limited to: agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, pregelatinized starch, other starches, clays, other algins, other celluloses, gums, and mixtures thereof. Lubricants useful in the pharmaceutical compositions and dosage forms of the present invention include, but are not limited to: calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerol, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oils (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, and mixtures thereof.
The scheme of the invention will be explained with reference to the examples. It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention. The examples do not specify particular techniques or conditions, and are performed according to techniques or conditions described in literature in the art or according to the product specification. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
Example 1
Respectively culturing the pancreatic islet beta cells and pancreatic precursor cells in a suspension manner for 6 days by using CEP-3379, TG101209 and TG101348, then culturing for 8 days again, and detecting Insulin precursors (Proinulin, Insulin (Insulin), C-Peptide (C-Peptide) and NKX6.1, which are specific molecular markers of the pancreatic islet beta cells, by using a flow cytometer, wherein NKX6.1 is a protein expressed by an NKX6.1 gene in the pancreatic islet beta cells, and the specific experimental process is as follows:
1. method for differentiating pancreatic precursor cells into pancreatic endocrine precursor cells
The differentiation of pancreatic precursor cells into pancreatic endocrine precursor cells was verified by suspension culture for 6 days. CEP-3379, TG101209 and TG101348 were added to the culture medium containing pancreatic precursor cells, respectively, wherein the working concentration of CEP-3379 (i.e., the concentration in the culture medium) was 0.05. mu.M, the working concentration of TG101209 was 0.5. mu.M and the working concentration of TG101348 was 0.2. mu.M.
The suspension culture plate is a low-adsorption six-well plate (Beaverbeio, Cat # 40406). The culture medium used was: DMEM (Gibco, Cat # C11965500CP) +50X vitamin A free B-27(Gibco, Cat #12587-010) + 1% glutamate (Glutamax, Invitrogen, Cat #35050079) +0.25mM 2 phosphorus-L-ascorbic acid trisodium salt (Sigma Aldrich; Cat #49752) + 10. mu.M ALK5 inhibitor II (Enzo Life Sciences; Cat # ALX-270-4)45-M100), Triiodo-L-thyronine (5-Triiodo-L-thyronine) (Sigma, Cat # T2877) +0.1 μ M γ -Secretase inhibitor XX (EMD Millipore; cat #565789-1 MG).
Wherein, the culture conditions are as follows: 37 ℃ and 5% CO 2 And 88% humidity at 90 rpm.
The pancreatic endocrine precursor cells are obtained by the suspension culture, and then CEP-3379, TG101209 or TG101348 are continuously added for suspension culture to promote the pancreatic endocrine precursor cells to differentiate into islet beta cells.
2. Method for differentiating pancreatic endocrine precursor cells into pancreatic beta cells
The differentiation of pancreatic endocrine precursor cells into islet beta cells was verified using a suspension culture method. The pancreatic endocrine precursor cells were cultured in the following medium for 8 days to examine whether or not the pancreatic endocrine precursor cells differentiated into islet beta cells.
Wherein, the suspension culture plate is a low-adsorption six-hole plate (Beaverdio, Cat # 40406). The media used were: DMEM (Gibco, Cat # C11965500CP) +50X vitamin A free B-27(Gibco, Cat #12587-010) + 1% Glutamax (Invitrogen, Cat #35050079) + 10. mu.M ALK5 Inhibitor II (Enzo Life Sciences; Cat # ALX-270-445-M100). Flow cytometry detection shows that the islet beta cells can be obtained after 4-8 days of culture.
3. Flow cytometry detection method
The detection method of Insulin precursor (Proinulin), Insulin (Insulin), C Peptide (C-Peptide) and NKX6.1 which are specific molecular markers of the islet beta cells is as follows:
the differentiated cell pellet was digested with preheated Accutase cell digest cell lysate (Sigma, A6964-100ML) for 5-10 minutes into single cells, washed once with PBS, and fixed with 200. mu.L of fixed permeate (BD; Cat #554722) for 20 min.
The Wash solution was then infiltrated with 200. mu.L of BD (BD Perm/Wash) TM Buffer, Cat #554723) were washed twice and incubated at 4 ℃ for 2 hours with primary antibody, which was formulated as follows: 200 μ L BD penetrating Wash (BD Perm/Wash) TM Buffer) +1 μ L of murine anti-NKX6.1 (mouse anti-NKX6.1, DSHB; cat # F55a12) +1 μ L rabbit C peptide antibody (Rat C-pep antibody, DSHB; cat # GN-ID 4).
After the incubation was completed, 200. mu.L of BD was used to permeate the Wash solution (BD Perm/Wash) TM Buffer) and incubated with a fluorescent secondary antibody at 4 ℃ for 1 hour in the dark, wherein the fluorescent secondary antibody was formulated as follows: 200 μ L BD Perm/Wash TM Buffer + monkey anti-rabbit Antibody (Donkey anti-Rat IgG (H + L) Secondary Antibody) with the fluorescent dye Alexa Fluor 488(Life Technologies; Cat # A-21208) + Alexa with the fluorescent dye
Figure BDA0003730619140000091
647 monkey anti-Mouse Antibody (Donkey anti-Mouse IgG (H + L) subcordary Antibody) to conjugate (Life Technologies; Cat # A-31571).
After the incubation was completed, 200. mu.L of BD Perm/Wash was used TM Buffer twice, then 200. mu.L BD Perm/Wash TM Buffer was resuspended, transferred to a flow tube, detected using a flow cytometer (BD Calibur), and flow results analyzed using FlowJo software.
The results of the experiment are shown in FIG. 1. As can be seen from FIG. 1, compared with a control, the addition of the chemical small molecule inhibitor related to the JAK2 signal pathway can effectively improve the expression levels of the specific molecular markers of the islet beta cells, namely, the Proinsulin/Insulin/C-Peptide and NKX6.1 (mainly shown as the content and fluorescence intensity of the molecular marker in Q2 are enhanced). As inhibitors of JAK2, CEP-33779, TG101209 and TG101348 were all effective in increasing differentiation efficiency of pancreatic endocrine precursor cells directed to islet β cells, fig. 1.
Meanwhile, the proportion of islet beta cells generated by the JAK2 inhibitor in the process of promoting the directional differentiation of pancreatic endocrine precursor cells into islet beta cells is remarkably analyzed, and the proportion is shown in figure 2. Wherein, both TG101209 and TG101348 can obviously improve the proportion of islet beta cells.
Furthermore, we investigated the case where addition of TG101348 at different time nodes induced pancreatic endocrine precursor cells to produce islet β cells. Taking the TG101348 as an example, as shown in FIG. 3, the ordinate in FIG. 3 represents different treatment times, wherein "1" represents one day of the treatment with the addition of TG101348, "1-2" represents the treatment with the addition of TG101348 on the first day, and the treatment is continued for two days, and the analogy from "1-3" represents the treatment with the addition of TG101348 on the first day, and the treatment is continued for three days. Similarly, "2-5" represents the addition of TG101348 on the next day until the fifth day. As can be seen from the results shown in FIG. 3, it was found that the highest proportion of islet beta cells was obtained by treating with TG101348 continuously for 6 days.
Example 2
We also evaluated the effect of JAK2 inhibitors on insulin cleavage, using as a control the absence of JAK2 inhibitor. The insulin shearing detection method used was as follows:
the differentiated cell pellet was digested with preheated Accutase cell digest cell separation (Sigma, A6964-100ML) for 5-10 minutes into single cells, washed once with PBS buffer, and fixed with 200. mu.L of fixed permeate (Fixation and Permeabilization Solution) for 20 min.
The Wash solution was then infiltrated with 200. mu.L of BD (BD Perm/Wash) TM Buffer) and incubated for 2 hours at 4 ℃ with primary antibody, wherein the primary antibody was formulated as follows: 200 μ L BD penetrating Wash solution (BD Perm/Wash) TM Buffer) +1 μ L of murine anti-NKX6.1 (mouse anti-NKX6.1, DSHB; cat # F55a12) +1 μ L of porcine anti-C Peptide antibody (Pig anti-C Peptide antibody, Abcam; cat # ab 30477).
After the incubation was completed, 200. mu.L of BD Perm/Wash was used TM Buffer washing twice, and incubating for 1 hour at 4 ℃ in the dark by using a fluorescent secondary antibody, wherein the fluorescent secondary antibody is prepared as follows: 200 μ L BD Perm/Wash TM Buffer + Alexa with fluorescent dye
Figure BDA0003730619140000101
Donkey anti-Mouse Antibody (Donkey anti-Mouse IgG (H + L) subcordary Antibody) to 488conjugate (Life Technologies; Cat # A-21202) + Alexa with fluorescent dye
Figure BDA0003730619140000102
647 Goat anti-Pig Antibody (Goat anti-Guinea Pig IgG (H + L) subcordary Antibody) to conjugate (Life Technologies; Cat # A-21450).
After the incubation was completed, 200. mu.L of BD Perm/Wash was used TM Buffer twice, then 200. mu.L BD Perm/Wash TM Buffer was resuspended, transferred to a flow tube, detected using a flow cytometer (BD Calibur), and flow results analyzed using FlowJo software.
As shown in fig. 4 and 5, it was found that, compared with the control, the addition of TG101348 not only increases the production rate of islet β cells, but also promotes the cleavage of proinsulin to insulin, so that the differentiated islet β cells more tend to functionally mature islet β cells.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.
Although embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are exemplary and not to be construed as limiting the present invention, and that changes, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.

Claims (13)

  1. Use of a JAK2 inhibitor for the preparation of an islet beta cell inducing differentiation agent.
  2. 2. The use of claim 1, wherein said islet beta cell inducing differentiation agent promotes differentiation of pancreatic endocrine precursor cells or pancreatic precursor cells into islet beta cells.
  3. 3. The use according to claim 1, wherein the JAK2 inhibitor is a JAK2 protein kinase inhibitor.
  4. 4. The use according to claim 3, wherein the JAK2 protein kinase inhibitor is selected from at least one of CEP-33779, TG101209, TG 101348.
  5. 5. An islet beta cell induction-differentiation agent comprising a JAK2 inhibitor, DMEM, vitamin a-free B-27, glutamate, and an ALK5 inhibitor.
  6. 6. The islet beta cell induction-differentiation agent according to claim 5, further comprising trisodium phosphate-L-ascorbate, triiodo-L-thyronine and gamma-secretase inhibitors.
  7. 7. An islet beta cell inducing differentiation agent according to claim 5, wherein said JAK2 inhibitor is a JAK2 protein kinase inhibitor.
  8. 8. The islet beta cell inducing differentiation agent according to claim 7, wherein said JAK2 protein kinase inhibitor is selected from at least one of CEP-33779, TG101209, TG 101348.
  9. 9. The islet beta cell inducing differentiation agent according to claim 5, further comprising other pharmaceutically acceptable carriers and/or excipients.
  10. 10. A pharmaceutical composition for preventing and/or treating diabetes mellitus, which comprises the islet beta cell-inducing differentiation agent according to any one of claims 5 to 9.
  11. 11. A method of promoting differentiation of cells into islet beta cells, comprising:
    contacting and culturing the cells with a JAK2 inhibitor for 5-7 days so as to obtain islet beta cells;
    the cells are pancreatic endocrine precursor cells or pancreatic precursor cells.
  12. 12. The method of claim 11, wherein the JAK2 inhibitor is a JAK2 protein kinase inhibitor.
  13. 13. The method according to claim 12, wherein the JAK2 protein kinase inhibitor is selected from at least one of CEP-33779, TG101209, TG 101348.
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