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Pharmaceutical composition as well as preparation method and application thereof

A composition and drug technology, applied in the field of pharmaceutical compositions and their preparation, can solve problems such as unpublished pulmonary degenerative diseases, and achieve the purpose of preventing lung cell apoptosis, inhibiting the secretion of inflammatory cytokines, and promoting the secretion of lung regeneration cytokines. Effect

Pending Publication Date: 2021-02-05
源铭(上海)生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no public record about the use of mesenchymal stem cells and their exosome pharmaceutical composition in the prevention or treatment of pulmonary degenerative diseases including chronic obstructive pulmonary disease

Method used

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  • Pharmaceutical composition as well as preparation method and application thereof
  • Pharmaceutical composition as well as preparation method and application thereof
  • Pharmaceutical composition as well as preparation method and application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0104] As the method for preparing CD31 and CD73 positive mesenchymal stem cells of the present invention, the method comprises the following steps:

[0105] Step 1: obtaining and culturing mesenchymal stem cells from the source tissue;

[0106] Step 2: dispersing the mesenchymal stem cells obtained in Step 1 to obtain a single cell suspension;

[0107] Step 3: Sorting and extracting CD31- and CD73-positive mesenchymal stem cells from the single-cell suspension obtained in Step 2.

[0108] These steps are described in detail below.

[0109] (1) Step 1: Obtain and culture mesenchymal stem cells from the source tissue. That is, a collection of mesenchymal stem cells obtained by isolating and culturing mesenchymal stem cells from tissues such as umbilical cord, fat, bone marrow, and placenta, preferably placental tissue, according to methods known to those skilled in the art.

[0110] The growth medium used for culturing mesenchymal stem cells of the present invention is a med...

Embodiment 1

[0199] Example 1: Preparation of CD31 and CD73 positive mesenchymal stem cells

[0200] Placentas were obtained from healthy pregnant women undergoing caesarean section, and informed consent statements were signed according to ethical requirements. After the placenta was collected, the decidua membrane and umbilical cord were peeled off, and the cotyledons were preserved. The cotyledons were then washed in Hanks buffered saline solution (HBSS) and then digested in a solution of 1 mg / mL collagenase I, 1 mg / mL Dnase-1 and 75 μg / mL Dispase at 37°C for 2 hours. After digestion, filter the single cell suspension through a 100 µm sieve and spin at 750 x g for 5 min. The supernatant was decanted and the cell pellet was then resuspended in erythrocyte lysis buffer and incubated in a 37°C incubator for 10 minutes. The suspension was then spun at 510 x g for 5 h min. The supernatant was then decanted, and the cell pellet was washed in Hanks buffered saline solution (HBSS) and spun ag...

Embodiment 2

[0203] Example 2: Detection of CD31 and CD73 positive mesenchymal stem cell surface markers

[0204]The third-generation mesenchymal stem cells cultured in Example 1 were taken, and the expressions of CD31, CD73, CD34 and HLA-DR were detected by flow cytometry. Reagents used are BD Pharmingen, USA, respectively FITC mouse anti-human CD31 (Cat#560984), PE mouse anti-human CD73 (Cat#550257, BD), FITC mouse anti-human CD34 (Cat#555821), FITC mouse anti-human HLA-DR (Cat#555811). Control antibodies are FICT mouseIgG1, k isotype (Cat#555909), PE mouse IgG2b, k isotype control (Cat#559529).

[0205] Staining and flow detection were carried out according to the experimental method provided by BD Pharmingen in the United States, and the results are shown in figure 2 .

[0206] The results showed that: the mesenchymal stem cells cultured in Example 1 were positive for expression of CD31 and CD73, and negative for expression of CD34\CD45 and HLA-DR.

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Abstract

The invention discloses a pharmaceutical composition as well as a preparation method and application thereof. In the application, the pharmaceutical composition comprises CD31- and CD73-positive mesenchymal stem cells and / or exosomes thereof, has functions of inhibiting inflammatory cytokine secretion, promoting lung regeneration cytokine secretion, inhibiting pulmonary fibrosis, preventing pulmonary cell apoptosis and the like, and can prevent or treat pulmonary degenerative diseases including chronic obstructive pulmonary disease (COPD).

Description

technical field [0001] The embodiments of the present invention relate to the field of pharmaceutical manufacturing, in particular to a pharmaceutical composition and its preparation method and application. Background technique [0002] Degenerative lung diseases, including chronic obstructive pulmonary disease (COPD), are a group of diseases with high morbidity and mortality worldwide. Chronic obstructive pulmonary disease (COPD) is an umbrella term for chronic bronchitis and emphysema, associated with a chronic inflammatory response of the lungs that causes airway abnormalities including airway constriction and distortion of the structure of the lung parenchyma. Patients typically experience a progressive decline in lung function, characterized by an intensified cough, shortness of breath, and sputum production. But not only that, the systemic immune changes in patients with chronic obstructive pulmonary disease (COPD) are characterized by an increase in the number of inf...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61K9/08A61P11/00A61P11/06
CPCA61K35/28A61K9/08A61P11/00A61P11/06A61K2300/00
Inventor 吴冬梅林峰周宁李淑亭伊希姆·E·托马斯
Owner 源铭(上海)生物技术有限公司
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