Chemical modification of recombinant hirudin with palmitic acid in mixed aqueous-organic solutions

Wang, Shuchang; Wang, Xudong; Teng, Xin‐Nan; Xiu, Zhilong

doi:10.1016/j.procbio.2018.11.022

Cited by 5 publications

(3 citation statements)

References 28 publications

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“…Moreover, the secondary structure of rhG-CSF in DMSO solvent could restore its correct form after being PEGylated and separated into the aqueous solution. Another previous study on the fatty chain modification of HV2 with PAL-BTA ( Wang et al, 2019 ) showed that both the two substrates had desirable solubility in mixed aqueous-DMSO solutions. Therefore, a mixed aqueous-DMSO solution was attempted to use as the solvent system for the modification of rhG-CSF with C15-MAL.…”

Section: Resultsmentioning

confidence: 98%

Preparation and Characterization of Site-Specific Fatty Chain-Modified Recombinant Human Granulocyte Colony Stimulating Factor

Wang

Liu

et al. 2022

Front. Bioeng. Biotechnol.

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The clinical use of recombinant human granulocyte colony-stimulating factor (rhG-CSF) is limited by its short serum half-life. In this study, a long-acting strategy for site-specific modification of rhG-CSF with 1-pentadecyl-1H-pyrrole-2,5-dione (C15 fatty chain-maleimide, C15-MAL) was studied in mixed DMSO-aqueous solutions. The factors influencing the conjugation reaction were investigated and optimized, and a high yield of the desired product (C15-rhG-CSF) was achieved. Subsequently, C15-rhG-CSF product was efficiently purified using preparative liquid chromatography, and further characterized. Circular dichroism spectroscopy analysis showed that the secondary structure of C15-rhG-CSF had no significant difference from unmodified rhG-CSF. C15-rhG-CSF retained 87.2% of in vitro bioactivity of unmodified rhG-CSF. The pharmacokinetic study showed that the serum half-life of C15-rhG-CSF in mice was 2.08-fold longer than that of unmodified rhG-CSF. Furthermore, C15-rhG-CSF by single-dose subcutaneous administration showed better in vivo efficacy than those of both PEG10k-rhG-CSF by single-dose administration and rhG-CSF by multiple doses administration. This study demonstrated the potential of C15-rhG-CSF being developed into a novel drug candidate as well as an efficient process for the development of long-acting protein and peptide drugs.

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Section: Resultsmentioning

confidence: 98%

Preparation and Characterization of Site-Specific Fatty Chain-Modified Recombinant Human Granulocyte Colony Stimulating Factor

Wang

Liu

et al. 2022

Front. Bioeng. Biotechnol.

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“…44 The half-life of hirudin could be prolonged via structural modification such as the incorporation of D-amino acids, noncanonical amino acids, conjugation with fatty acids, and others. [45][46][47][48] In the present work, we suggested several modifications of the desulfo-hirudin molecule to enhance its affinity for thrombin that might enable the reduction of hirudin dose in the clinic and, therefore, the risk of bleeding. We introduced modifications that mimic sulfation on Tyr63, such as phosphotyrosine (pTyr), carboxymethylphenylalanine (CMF), and glutamate residue.…”

Section: Introductionmentioning

confidence: 93%

“…Also, the improved pharmacological properties were shown for encapsulated hirudin in the engineered nanogel, promoting its accumulation in the blood clots and its self‐regulatory release during the pathological proceeding of thrombus and alleviating adverse effects 44 . The half‐life of hirudin could be prolonged via structural modification such as the incorporation of D‐amino acids, noncanonical amino acids, conjugation with fatty acids, and others 45–48 …”

Section: Introductionmentioning

confidence: 99%

Tyrosine phosphorylation of recombinant hirudin increases affinity to thrombin and antithrombotic activity

Volkova,

Semenyuk

2023

Proteins

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Thrombin is one of the key enzymes of the blood coagulation system and a promising target for the development of anticoagulants. One of the most specific natural thrombin inhibitors is hirudin, contained in the salivary glands of medicinal leeches. The medicinal use of recombinant hirudin is limited because of the lack of sulfation on Tyr63, resulting in a 10‐fold decrease in activity compared to native (sulfated) hirudin. In the present work, a set of hirudin derivatives was tested for affinity to thrombin: phospho‐Tyr63, Tyr63(carboxymethyl)Phe, and Tyr63Glu mutants, which mimic Tyr63 sulfation and Gln65Glu mutant and lysine‐succinylated hirudin, which enhance the overall negative charge of hirudin, as well as sulfo‐hirudin and desulfo‐hirudin as references. Using steered molecular dynamics simulations with subsequent umbrella sampling, phospho‐hirudin was shown to exhibit the highest affinity to thrombin among all hirudin analogs, including native sulfo‐hirudin; succinylated hirudin was also prospective. Phospho‐hirudin exhibited the highest antithrombotic activity in in vitro assay in human plasma. Taking into account the modern methods for obtaining phospho‐hirudin and succinylated hirudin, they are prospective as anticoagulants in clinical practice.

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Surface functionalization of anticoagulation and anti-nonspecific adsorption with recombinant hirudin modification

Zheng

Dai

et al. 2022

Biomaterials Advances

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Chemical modification of recombinant hirudin with palmitic acid in mixed aqueous-organic solutions

Cited by 5 publications

References 28 publications

Preparation and Characterization of Site-Specific Fatty Chain-Modified Recombinant Human Granulocyte Colony Stimulating Factor

Preparation and Characterization of Site-Specific Fatty Chain-Modified Recombinant Human Granulocyte Colony Stimulating Factor

Tyrosine phosphorylation of recombinant hirudin increases affinity to thrombin and antithrombotic activity

Surface functionalization of anticoagulation and anti-nonspecific adsorption with recombinant hirudin modification

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