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Compositions and methods for modifying the surface of cells and methods of use

A composition and cell technology, applied in the described field, can solve the problems of cytotoxicity and damage to the normal function of cells

Inactive Publication Date: 2016-11-02
UNIV OF UTAH RES FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, cell surface modifications often cause severe cytotoxicity, or impair normal cell function

Method used

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  • Compositions and methods for modifying the surface of cells and methods of use
  • Compositions and methods for modifying the surface of cells and methods of use
  • Compositions and methods for modifying the surface of cells and methods of use

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Experimental program
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preparation example Construction

[0091] a. Compound preparation

[0092] Certain compounds described herein may be commercially available. Others may require synthetic preparation. For the preparation of compounds of formula I, a phospholipid-PEG intermediate bearing a reactive functional group on the terminus of the PEG can be coupled to a targeting moiety.

[0093] For example, when the reactive functional group is NHS, the intermediate can be coupled to a targeting moiety with a reactive amino group to form an amide. When the reactive functional group is an amine, the intermediate can be coupled with a reactive isothiocyanate to form a thiocarbamate. When the reactive functional group is azide, the intermediate can be coupled with a targeting moiety with a reactive nitrile or reactive alkyne group to form a tetrazole or triazole. When the reactive functional group is maleimide, the intermediate can be coupled to a targeting moiety with a reactive thiol group to form a bond between the sulfur of the thio...

Embodiment 1

[0122] compound synthesis

[0123] General experimental and analytical details

[0124] Functionalized DMPE-PEG (3.4 kDa and 5 kDa) were purchased from Nanocs Corporation (New York, NY). Fluorescein isothiocyanate (FITC) and rhodamine were purchased from ThermoScientific (Rockford, IL).

[0125] in CDCl with 0.03% TMS as internal standard 3 Medium or DMSO-d 6 Recorded on a Bruker AM 400 spectrometer (operated at 400 and 101 MHz, respectively) or a Bruker AVIII spectrometer (operated at 500 and 126 MHz, respectively) 1 H and 13 C NMR spectra. Chemical shifts (δ) are in parts per million (ppm) and coupling constants (J) are in Hertz (Hz). Spin multiplicity is reported as s = singlet, br.s = broad singlet, d = doublet, t = triplet, q = quartet, dd = double doublet, and m = multiplet. LCMS analysis was performed on an Agilent 1200 RRL chromatograph with photodiode array UV detection and an Agilent 6224 TOF mass spectrometer. Chromatography used the following parameters: Wa...

Embodiment 2

[0139] MSC modification

[0140] General Experimental Procedure : Culture-expanded MSCs were harvested and suspended in phenol red-free medium. DMPE-PEG or DMPE-PEG-CXCR4 was added directly to the cell suspension. At predetermined time points, MSCs were harvested, washed, and suspended in HBSS. To optimize the amount of DMPE-PEG, different amounts of DMPE-PEG were mixed with 750,000 MSCs.

[0141] Effect of PEG size on MSC surface modification : Two DMPE-PEG compositions with different PEG sizes, namely 5kDa and 3.4kDa, were labeled with FITC for fluorescence detection, and the DMPE-PEG-FITC compounds were subsequently incubated with MSCs in suspension. The ability of compounds to bind to the surface of MSCs was assessed by confocal microscopy and FACS analysis (see figure 1 ). The results show that the incorporation of 5kDa PEG into the composition is superior to the use of 3.4kDa PEG. All subsequent experiments described herein used DMPE-PEG compositions with 5 kDa ...

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Abstract

Described herein are compounds, compositions and methods for modification of the surface of a living cell with a therapeutically relevant targeting moiety. Also described herein are methods for treating disease states, such as acute myocardial ischemia or infarction, with said compositions, in a subject.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Application 61 / 929,430, filed January 20, 2014, and is hereby incorporated by reference in its entirety. Background technique [0003] Cellular therapies such as stem cell therapy offer great promise for treating physical ailments, diseases and tissue defects. A significant obstacle to the effective delivery of cell therapies is the inability to efficiently target large numbers of living cells to target tissues. Systemic infusion is generally desired because it minimizes the invasiveness of the cell therapy and maximizes the practical aspects of repeated dosing. It also allows the cells to mimic the natural cellular trafficking process and helps ensure that the cells remain close to oxygen and nutrient-rich blood vessels. However, cells often exhibit poor homing ability or lose their homing ability after ex vivo culture expansion. [0004] For example, many preclinica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/127G01N33/53
CPCC12N5/0006C12N5/0663A61K2035/124A61K49/0041A61K49/0047A61K49/0084A61K47/64A61K47/6911A61K47/6425A61K35/28C12N5/0662A61K47/60
Inventor 元映旭大卫·A·布尔阿米特·N·帕特尔
Owner UNIV OF UTAH RES FOUND
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