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. 2022 Mar 16:10:859133.
doi: 10.3389/fchem.2022.859133. eCollection 2022.

Synthesis and Characterization of Phenylboronic Acid-Modified Insulin With Glucose-Dependent Solubility

Nai-Pin Lin  1   2 , Nan Zheng  2 , Landa Purushottam  1 , Yi Wolf Zhang  1   2 , Danny Hung-Chieh Chou  1   2
Affiliations

Synthesis and Characterization of Phenylboronic Acid-Modified Insulin With Glucose-Dependent Solubility

Nai-Pin Lin et al. Front Chem. .

Abstract

Glucose-responsive insulin represents a promising approach to regulate blood glucose levels. We previously showed that attaching two fluorophenylboronic acid (FPBA) residues to the C-terminal B chain of insulin glargine led to glucose-dependent solubility. Herein, we demonstrated that relocating FPBA from B chain to A chain increased the baseline solubility without affecting its potency. Furthermore, increasing the number of FPBA groups led to increased glucose-dependent solubility.

Keywords: glucose responsiveness; insulin; insulin modification; peptide; phenylboronate.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
(A) Schematic mechanism of glucose-responsive insulin. (B) Design of FPBA-conjugated glucose-responsive insulin.
SCHEME 1
SCHEME 1
Synthesis of FPBA-conjugated insulin analogs.
FIGURE 2
FIGURE 2
(A) Solubility of insulin analogs in pH 7.4 PBS. Data are expressed as mean ± SD (n = 4 per group). (B) Solubility of insulin analogs in pH 7.4 PBS with different glucose concentrations. Data are expressed as mean ± SD (n = 4 per group). (C) In vitro activity of insulin analogs by using activated pAKT levels as measurements. Data are expressed as mean ± SD (n = 4 per group). EC50 was calculated by Prism 9 (GraphPad Software, CA, United States) with nonlinear regression curve fitting of dose-response asymmetric equation.
SCHEME 2
SCHEME 2
Conjugation of FPBA to peptides by Alloc strategy.
SCHEME 3
SCHEME 3
Synthesis of A chain FPBA-conjugated insulin analogs.
FIGURE 3
FIGURE 3
(A) Solubility of insulin analogs in pH 7.4 PBS. Data are expressed as mean ± SD (n = 4 per group). The statistical comparison of the two groups was evaluated by the unpaired two-tailed Student’s t-test. *p < 0.001. (B) Solubility of insulin analogs in pH 7.4 PBS with different glucose concentrations. Data are expressed as mean ± SD (n = 4 per group). (C) In vitro activity of insulin analogs by using activated pAKT levels as measurements. Data are expressed as mean ± SD (n = 4 per group). EC50 was calculated by Prism 9 (GraphPad Software, CA, United States) with nonlinear regression curve fitting of dose-response asymmetric equation.
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