CN104693075A - p18 small-molecule inhibitor and application thereof in human hematopoietic stem cell in-vitro amplification - Google Patents
p18 small-molecule inhibitor and application thereof in human hematopoietic stem cell in-vitro amplification Download PDFInfo
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- CN104693075A CN104693075A CN201510081641.9A CN201510081641A CN104693075A CN 104693075 A CN104693075 A CN 104693075A CN 201510081641 A CN201510081641 A CN 201510081641A CN 104693075 A CN104693075 A CN 104693075A
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- LHFSIYNKTKMPPE-UHFFFAOYSA-N N=[O]C(c(cc1)ccc1S(NC1CCCCC1)(=O)=O)=O Chemical compound N=[O]C(c(cc1)ccc1S(NC1CCCCC1)(=O)=O)=O LHFSIYNKTKMPPE-UHFFFAOYSA-N 0.000 description 1
- PTCSSXYPZOFISK-UHFFFAOYSA-N OC(c(cc1)ccc1S(Cl)(=O)=O)=O Chemical compound OC(c(cc1)ccc1S(Cl)(=O)=O)=O PTCSSXYPZOFISK-UHFFFAOYSA-N 0.000 description 1
- XGOXPTXOQXHIDL-UHFFFAOYSA-N OC(c(cc1)ccc1S(NC1CCCCC1)(=O)=O)=O Chemical compound OC(c(cc1)ccc1S(NC1CCCCC1)(=O)=O)=O XGOXPTXOQXHIDL-UHFFFAOYSA-N 0.000 description 1
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Abstract
The invention relates to a p18 small-molecule inhibitor and application thereof in human hematopoietic stem cell in-vitro amplification. The structure is disclosed in the specification. The invention also relates to a preparation method and application of the p18 small-molecule inhibitor. The p18 small-molecule inhibitor has favorable promotion action on self-renewal of human hematopoietic stem cells, and can provide a new method for subsequent clinical treatment.
Description
Technical field
The present invention relates to field of medicaments, particularly p18 micromolecular inhibitor and the purposes in the amplification in vitro of human cord blood source hemopoietic stem cell thereof.
Background technology
Stem cell becomes one of effective way of clinical treatment various diseases because it has the ability of self and Multidirectional Differentiation, and this also represent the arrival of regenerative medicine New Times.Wherein hemopoietic stem cell research starts the earliest, studies the most deeply and be most widely used.The malignant hematologic diseases such as application bone marrow transplantation therapy leukemia have the history of more than 50 year.But hemopoietic stem cell quantity is lower in marrow, be difficult to effectively carry out hematopoietic reconstitution after transplanting in patient body, this also becomes a bottleneck of stem cell clinical application.
Hemopoietic stem cell has multiple choices in vivo, such as self, differentiation, migration (going back to the nest), tranquillization, apoptosis etc., has the different fate of a very complicated signal path network adjustment hemopoietic stem cell in these different selections behind.Such as Wnt, Notch, Shh/BMP, TGF-β, IGF etc. grow relevant Function protein, and the chromatin correlation factors such as Bmi, the transcription factors such as HoxB, NF κ, the cyclins etc. such as INK4, KIP, PTEN all take part in this Signal Regulation network.For hemopoietic stem cell, its of paramount importance characteristic is self.Existing multinomial research is at present pointed out, the self of hemopoietic stem cell regulates with the adjustment of cell cycle closely bound up.Entering the cell cycle is the final step that hemopoietic stem cell carries out self, therefore cell cycle, and especially the adjustment of G1 phase may produce vital effect to the self of hemopoietic stem cell.
Cell cycle regulates and controls mainly through the activity change of continuous print cell cycle protein dependent kinase (CDKs), and the activity of CDK must depend on the contents level of Cyclin.In vivo, except the phosphorylation/dephosphorylized regulation and control of Cyclin and catalytic subunit, CDK is to a great extent also by the regulation and control of CKIs (cyclin-dependent kinase inhibitors).Two class lower molecular weight family proteins in current known CKI, Cip/Kip and INK4 can react with CDK, and capable of inhibiting cell by the G1 phase.Wherein, Cip/Kip family, mainly comprises p21, p27 and p57, can react with a lot of Cyclin-CDK mixture; INK family, mainly comprises p16, p15, p18 and p19, can specific suppression CDK4,6.
Increasing research shows, for other CKI, the self regulating and controlling effect of p18 to hemopoietic stem cell is more powerful.Such as, the disappearance of p18 can cause the remarkable rising of mouse hemopoietic implantation rate, and this kind of effect is that the ratio divided by improving hemopoietic stem cell symmetry self realizes, compared to the proliferate efficiency improving merely hemopoietic stem cell in the past, improve differentiation and exhaustion that the division of its symmetry can avoid causing hemopoietic stem cell.Therefore, p18 is the INK albumen of the uniqueness of a specific effect hemopoietic stem cell self.
Current amplifying candidate stem cell in vitro mainly relies on and add the cytokine such as serum and SCF, TPO, Flt3L in substratum, but expanding effect is still undesirable, and hemopoietic stem cell just breaks up in vitro very soon, exhaustion.Once adopted in research before add cytokine profiles liquid nutrient medium, with stroma cell co-cultivation or the method such as to cultivate in bio-reactor with proliferation of hematopoietic stem/progenitor cell in vitro, but aforesaid method still can not obtain the sufficient hemopoietic stem cell with transfer ability to carry out clinical treatment.
Micromolecular compound shows the advantage of its uniqueness in then cultivating in vitro.First, the action effect of compound by regulating the structure of compound, concentration changes, its process is simply controlled; Secondly, micromolecular compound is more stable compared to its character such as cytokine or transcription factor, the process in early stage of compound and administration very convenient.Therefore, in the method for conventional amplification hemopoietic stem cell, add micromolecular compound, adopt micromolecular compound to substitute the materials such as existing serum, cytokine even completely, may play a significant role to vitro culture hemopoietic stem cell.
The present invention is intended to the obtained a kind of new p18 micromolecular inhibitor of research, and by this p18 micromolecular inhibitor vitro culture human hematopoietic stem cell, to reach the object of the amplification in vitro of hemopoietic stem cell.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, provides a kind of p18 micromolecular inhibitor and the purposes in human hematopoietic stem cell amplification in vitro thereof.
The technical solution used in the present invention is:
A kind of p18 micromolecular inhibitor, its structure is:
Present invention also offers the preparation method of this p18 micromolecular inhibitor, comprise the steps:
Under condition of ice bath, by hexahydroaniline be dissolved in methylene dichloride to carboxylphenylsulphonyl chloride, add triethylamine, stir 1 hour, after rising to room temperature, continue to stir, TLC monitors after reaction completes, rotary evaporation removing reaction soln, and residue purified, obtains intermediate;
Under condition of ice bath, be dissolved in water, add sodium hydroxide by gained intermediate, after rising to room temperature, stir 30 minutes, gained reaction solution vacuum-freeze-dry, obtains finished product.
The reaction equation related to is:
Present invention also offers the purposes of this p18 micromolecular inhibitor in human hematopoietic stem cell amplification in vitro.
Particularly, in described purposes, the concentration of p18 micromolecular inhibitor in amplification culture medium is 10-50nM.
Particularly, present invention also offers a kind of human hematopoietic stem cell amplification in vitro method, comprise the steps:
By the human cord blood CD 34 of fresh separated
+cell use amplification culture medium (StemSpan SFEM substratum+[100ng/mL] SCF/TPO/Flt3L/IL-6) resuspended to cell concn be 5 × 10
4cell/mL, is then laid in 96 orifice plates, and every hole is 190 μ L cell suspensions, and subsequently to the p18 micromolecular inhibitor wherein adding 10 μ L and use amplification culture medium of the same race diluted, the final concentration of p18 micromolecular inhibitor is 20nM.At 37 DEG C, 5%CO
2cultivate 7 days in constant incubator.
The beneficial effect that the present invention has:
P18 micromolecular inhibitor of the present invention has good promoter action for human hematopoietic stem cell self, can be the method that clinical treatment in the future provides new.
Accompanying drawing explanation
Fig. 1: cell phenotype ratio change statistical study figure after vitro culture in embodiment 3;
Fig. 2: cell phenotype absolute quantity change statistical study figure after vitro culture in embodiment 3;
Fig. 3: embodiment 4 cobble sample region forms cell Poisson statistics distribution plan;
Fig. 4: in embodiment 5, flow cytometer showed detects and transplants people CD45 in rear marrow
+cell content scatter diagram;
Fig. 5: in embodiment 5, statistical study derives from the change of the cell content of people in each group mouse bone marrow cells after transplanting.
Embodiment
In order to understand the present invention, below in conjunction with specific embodiment, the invention will be further described, but do not limit protection scope of the present invention.
Embodiment 1
A kind of p18 micromolecular inhibitor, its structure is:
The preparation method of this p18 micromolecular inhibitor, comprises the steps:
Under condition of ice bath, by hexahydroaniline (495 milligrams, 5.0 mmoles) with to carboxylphenylsulphonyl chloride (1100 milligrams, 5 mmoles) be dissolved in methylene dichloride (100 milliliters), add triethylamine (707 milligrams, 7 mmoles), stir 1 hour, after rising to room temperature, continue to stir, after TLC monitoring reaction completes, rotary evaporation removing reaction soln, the quick preparative column of resistates is purified, and obtains finished product (intermediate) 300 milligrams, yield 21%.
1H NMR(400MHz,DMSO-d6):δ7.94(d,J＝8.4Hz,2H),7.68(d,J＝8.4Hz,2H),7.51(bs,1H),2.90-2.91(m,1H),1.42-1.55(m,5H),0.99-1.11(m,5H).LC-MS(ESI):m/z 284.0(M+H)
+.
Under condition of ice bath, by (103 milligrams, above-mentioned intermediate, 0.36 mmole) be dissolved in 5 ml waters, add sodium hydroxide (14.56 milligrams, 0.36 mmole), after rising to room temperature, stir 30 minutes, gained reaction solution vacuum-freeze-dry, obtains finished product 100 milligrams, yield 90%.
Sodium 4-(N-cyclohexylsulfamoyl)benzoate(005A).
1H NMR(400MHz,DMSO-d6):δ7.74(d,J＝8.4Hz,2H),7.41(d,J＝6.8Hz,2H),3.60-3.61(m,1H),1.46-1.58(m,5H),0.99-1.16(m,5H).
LC-MS(ESI):m/z 283.1(M+H)
+。
Embodiment 2
A kind of human hematopoietic stem cell amplification in vitro method, comprises the steps:
By the human cord blood CD 34 of fresh separated
+cell use amplification culture medium (StemSpan SFEM substratum+[100ng/mL] SCF/TPO/Flt3L/IL-6) resuspended to cell concn be 5 × 10
4cell/mL, is then laid in 96 orifice plates, and every hole is 190 μ L cell suspensions, and subsequently to the p18 micromolecular inhibitor wherein adding 10 μ L and use amplification culture medium of the same race diluted, the final concentration of p18 micromolecular inhibitor is 20nM.At 37 DEG C, 5%CO
2cultivate 7 days in constant incubator.
Embodiment 3: cell phenotype analysis
Adopt flow cytometer detection CD34
+, CD34
+cD49f
+the ratio of cell and absolute quantity are verified for the effect of p18 micromolecular inhibitor amplifying candidate stem cell in vitro, and result display p18 micromolecular inhibitor is obvious for the amplification in vitro effect of hemopoietic stem cell.
Flow cytometer showed detects p18 micromolecular inhibitor to the experimental technique of human hematopoietic stem cell amplification in vitro effect:
1. umbilical cord blood HES (5:1) sedimented red cell will collected, after 60 minutes, collects supernatant liquid, with the ACK of 4-5 times of volume in 37 DEG C of water-bath splitting erythrocyte 15 minutes, and centrifugal 1800rpm, 10 minutes;
2. abandon supernatant, observe erythrocyte splitting situation, by cell harvesting in 1 50mL centrifuge tube, with 40ml ACK again in 37 DEG C of water-bath cracking 15 minutes, centrifugal 1800rpm, 10 minutes;
3. by cell 20ml PE buffer solution one time, filter, centrifugal 1500rpm, 8 minutes.Count cell simultaneously, add CD34 magnetic bead according to MNCs number, fully the rear 4 DEG C of lucifuges of mixing hatch 30 minutes;
4. after hatching end, every effective 20mlPE damping fluid washes one time, if there is precipitation, needs to filter, centrifugal 1500rpm, 5 minutes.Abandon supernatant, resuspended with 1mlPE damping fluid;
5. LS post is installed, moistens post 3 times with 1mlPE damping fluid.Dropwise add cell suspension, pass through in magnetic field;
6. post is washed with 2mlPE damping fluid, 2 times;
7. pillar is shifted out magnetic field, be placed on collection tube, add 3mlPE damping fluid, release cell fast with piston, get 10ul counting.
8. by obtain cell amplification culture medium (StemSpan SFEM substratum+[100ng/mL] SCF/TPO/Flt3L/IL-6) re-suspended cell to concentration be 5.0 × 10
4cells/mL, is laid in 96 orifice plates, and immediately add the testing compound (p18 micromolecular inhibitor) of proper concn, control group does not then add p18 micromolecular inhibitor;
9. cell is in 5%CO
2, cultivate 7 days in 37 DEG C of cell culture incubators;
10., after 7 days, flow cytometer showed detects the ratio of CD34 and CD49f;
According to the numerical value of each group's cell quantity drawn and ratio, Prism5 software is adopted to carry out statistical study.
Interpretation of result:
CD34 can be significantly improved in cultivating in vitro from Fig. 1,2, p18 micromolecular inhibitor 005A
+cD49f
+(005A group (20nM concentration) is 30.83% to the ratio of cell, control group is 19.87%), in the rear 005A group of process during 20nM concentration, it is about 1.5 times of control group, and has significant difference (p<0.05).Meanwhile, under this concentration, after p18 micromolecular inhibitor 005A process, CD34
+cD49f
+the absolute quantity of cell also improves about 1.4 times, and (005A group is 5509/10
4cells, control group is 3831/10
4cells, p<0.05).From above result, the level of hemopoietic stem cell after the process of p18 micromolecular inhibitor, really can be significantly improved.
Embodiment 4: cells in vitro functional analysis
Adopt pebbles sample region to form cell analysis (Cobblestone area forming cells analysis) and carry out functional analysis to the human hematopoietic stem cell through the process of p18 micromolecular inhibitor, result display p18 micromolecular inhibitor extracorporeal treatment can significantly improve the level of hemopoietic stem cell.
1. recovery M2-10B4 cell, with Secondary Culture base (high sugared 1640+10%FBS) re-suspended cell, and carries out cell counting;
2. use collagen protein bag by 96 orifice plates.Every hole about adds 50 μ L collagen solutions, and jog orifice plate makes it be laid at the bottom of plate.Partly uncap and be placed in super clean bench, room temperature dries at least 1h;
3. stroma cell is irradiated; Basis of microscopic observation stroma cell in good condition and when reaching required cell count digest collecting cell; Use Secondary Culture base to wash cell twice, and carry out cell counting; Use long period substratum (H5100+10
-6m HC) re-suspended cell, make cell concn be about 10
6-10
8/ mL, irradiation dose is 8000cGy (X-ray);
4. use in PBS damping fluid or long period substratum and the floating acid on collagen protein surface;
5. irradiate rear centrifugal collecting cell, and carry out cell counting.Use long period substratum re-suspended cell, make its concentration be 1.25 × 10
4/ mL;
6. cell being inoculated in collagen protein has wrapped in 96 orifice plates of quilt, and every hole adds 100 μ L M2-10B4 cell suspensions, in incubator (37 DEG C, 5%CO
2) middle cultivation, just can spread confluent monolayer cells (i.e. cell to be measured) after at least spending the night.
7. collect cell to be measured, herein cell to be measured detailed preparation method with reference to step in embodiment 3 1.-9..Carry out cell counting, then use long period substratum re-suspended cell; Remove the substratum of in orifice plate 90% carefully, note not blotting or stirring stromal cells layers;
8. every hole adds upper strata cell to be measured by calculating concentration, 100 μ L/ holes;
9. half amount changes liquid (not containing p18 micromolecular inhibitor in fresh culture) weekly, does not carefully stir stromal cells layers when changing liquid.
10. after 5 weeks, the positive hole count of counting containing pebbles sample region, and utilize Poisson's distribution to add up, adopt Prism5 software to draw.
Interpretation of result:
As shown in Figure 3, after p18 micromolecular inhibitor 005A (20nM) process, in cell, primitive hematopoietic cell ratio obviously rises, and is 1/397 (1/313 ~ 1/503), control group is then 1/1031 (1/745 ~ 1/1425), p=0.0174.Therefore can think that after p18 micromolecular inhibitor 005A process, the cell proportion that can form primitive hematopoietic region is increased significantly, therefore think that 005A can amplifying candidate stem cell in vitro.
Embodiment 5: animal model test
Employing immune deficiency mouse model is the gold standard of the hematopoietic reconstitution ability detecting hemopoietic stem cell, the long term hematopoietic ability of transplanted cells can be detected by transplantation experiments, found by this experiment, the hematopoietic reconstitution ability of not damaging cells after p18 micromolecular inhibitor 005A process, and effectively can improve the ratio of hematopoietic reconstitution.
1. by the fresh human cord blood CD 34 got
+cell goes out CD34 through airflow classification
+38
-45RA
-90
+cell;
2. the CD34 of some amount is got
+38
-45RA
-90
+cell is transplanted in Day0, as not cultivating group contrast;
3. the CD34 will obtained
+38
-45RA
-90
+cell amplification culture medium (StemSpan SFEM substratum+[100ng/mL] SCF/TPO/Flt3L/IL-6) re-suspended cell to concentration is 5.0 × 10
4cells/mL, is laid in 96 orifice plates, and immediately add the testing compound (p18 micromolecular inhibitor) of proper concn, control group does not then add p18 micromolecular inhibitor; Subsequently in 5%CO
2, cultivate 7 days in 37 DEG C of cell culture incubators; Then transplant;
4. adopted mouse is 4-5 NOD-SCID mouse in age in week, and before transplanting, 8-10h carries out medial lethal dose (220cGy) irradiation;
5. transplant according to front 500 cells of a cultivation/mouse, injection system adopts one-sided shin bone pulp cavity to transplant;
6. after 12 weeks, put to death mouse, get bilateral marrow and carry out flow cytometer showed detection people CD45 cell proportion respectively.
Interpretation of result:
As shown in Figure 4, utilize above experimental technique, after transplanting, can successfully occur the cell mass deriving from people in mouse.And as shown in Figure 5, after p18 micromolecular inhibitor 005A process, the hematopoietic reconstitution ability of cell is significantly higher than control group, the cell proportion of people is derived from apparently higher than control group (p<0.05) in mouse bone marrow cells, be on close level with the reconstruction of hemopoietic stem cell when not cultivating, and by migrating in injection side marrow (IF) in non-injection side marrow (BM), and in control group mice, in non-injection side marrow, can substantially have no the cell (p<0.05) deriving from people.Meanwhile, after 005A process, the mouse number of elements of hematopoietic reconstitution level higher (>10%) is higher than does not cultivate group (005A group is 6/9, and do not cultivate group be that 2/8,005A group is about 2.67 times that do not cultivate group).Therefore can think, after vitro culture, cell does not lose the ability of its hematopoietic reconstitution, and has carried out effective self.
Claims (5)
1. a p18 micromolecular inhibitor, is characterized in that: its structure is:
2. the preparation method of p18 micromolecular inhibitor described in claim 1, is characterized in that: comprise the steps:
Under condition of ice bath, by hexahydroaniline be dissolved in methylene dichloride to carboxylphenylsulphonyl chloride, add triethylamine, stir 1 hour, after rising to room temperature, continue to stir, TLC monitors after reaction completes, rotary evaporation removing reaction soln, and residue purified, obtains intermediate;
Under condition of ice bath, be dissolved in water, add sodium hydroxide by gained intermediate, after rising to room temperature, stir 30 minutes, gained reaction solution vacuum-freeze-dry, obtains finished product.
3. the purposes of p18 micromolecular inhibitor in human hematopoietic stem cell amplification in vitro described in claim 1.
4. purposes according to claim 3, is characterized in that: in described purposes, and the concentration of p18 micromolecular inhibitor in amplification culture medium is 10-50nM.
5. a human hematopoietic stem cell amplification in vitro method, is characterized in that: comprise the steps: the human cord blood CD 34 of fresh separated
+cell use amplification culture medium (StemSpan SFEM substratum+[100ng/mL] SCF/TPO/Flt3L/IL-6) resuspended to cell concn be 5 × 10
4cell/mL, is then laid in 96 orifice plates, and every hole is 190 μ L cell suspensions, and subsequently to the p18 micromolecular inhibitor wherein adding 10 μ L and use amplification culture medium of the same race diluted, the final concentration of p18 micromolecular inhibitor is 20nM, at 37 DEG C, and 5%CO
2cultivate 7 days in constant incubator.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108235708A (en) * | 2017-12-29 | 2018-06-29 | 罗小霞 | Cultivating system and its method, the candidate stem cell and hematopoietic progenitor cells of amplifying candidate stem cell and/or hematopoietic progenitor cells |
| CN110669733A (en) * | 2019-11-05 | 2020-01-10 | 中国医学科学院血液病医院(中国医学科学院血液学研究所) | Application of chrysin in human hematopoietic stem cell in-vitro amplification |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1871342A (en) * | 2003-10-24 | 2006-11-29 | 程涛 | P18 in stem cell manipulations |
| WO2008050167A1 (en) * | 2006-10-27 | 2008-05-02 | Richter Gedeon Nyrt. | New phenylsulfamoyl benzamide derivatives as bradykinin antagonists |
| CN102317282A (en) * | 2009-02-12 | 2012-01-11 | 安斯泰来制药有限公司 | Hetero ring derivative |
-
2015
- 2015-02-13 CN CN201510081641.9A patent/CN104693075B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1871342A (en) * | 2003-10-24 | 2006-11-29 | 程涛 | P18 in stem cell manipulations |
| WO2008050167A1 (en) * | 2006-10-27 | 2008-05-02 | Richter Gedeon Nyrt. | New phenylsulfamoyl benzamide derivatives as bradykinin antagonists |
| CN102317282A (en) * | 2009-02-12 | 2012-01-11 | 安斯泰来制药有限公司 | Hetero ring derivative |
Non-Patent Citations (3)
| Title |
|---|
| SALVATORE FERLA ET AL.: "Small-Molecule Inhibitors of 25-Hydroxyvitamin D‑24-Hydroxylase （CYP24A1）：Synthesis and Biological Evaluation", 《JOURNAL OF MEDICINAL CHEMISTRY》 * |
| SALVATORE FERLA ET AL.: "Small-Molecule Inhibitors of 25-Hydroxyvitamin D‑24-Hydroxylase （CYP24A1）：Synthesis and Biological Evaluation", 《JOURNAL OF MEDICINAL CHEMISTRY》, vol. 57, 22 August 2014 (2014-08-22), pages 7702 - 7715 * |
| 张亚斌等: "探讨脐血造血干细胞移植在临床应用中的研究进展", 《中国医药指南》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108235708A (en) * | 2017-12-29 | 2018-06-29 | 罗小霞 | Cultivating system and its method, the candidate stem cell and hematopoietic progenitor cells of amplifying candidate stem cell and/or hematopoietic progenitor cells |
| CN108235708B (en) * | 2017-12-29 | 2020-12-01 | 诺未科技（北京）有限公司 | Culture system and method for expanding hematopoietic stem cells and/or hematopoietic progenitor cells, hematopoietic stem cells, and hematopoietic progenitor cells |
| CN110669733A (en) * | 2019-11-05 | 2020-01-10 | 中国医学科学院血液病医院(中国医学科学院血液学研究所) | Application of chrysin in human hematopoietic stem cell in-vitro amplification |
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