Pic1 variants with improved solubility and methods of using the same

A technology of complement and donor, applied in the field of PIC1 variant with improved solubility and its use, can solve the problem of DHTR not being recognized

Pending Publication Date: 2021-10-01
瑞尔塔控股有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A primary or anamnestic response, leading to DHTR, may occur 1 to 4 weeks after transfusion with RBCs bearing this antigen [34]
[0007] Many DHTRs are believed to be mild and self-limiting, making DHTR often unrecognized

Method used

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  • Pic1 variants with improved solubility and methods of using the same
  • Pic1 variants with improved solubility and methods of using the same
  • Pic1 variants with improved solubility and methods of using the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Example 1: Peptide Solubility

[0097]To assess the effect of sarcosine residues on the solubility and biological function of the base peptide IALILEPICCQERAA(PA), peptides were synthesized with sarcosine residue substitutions at all 15 positions. Also included are peptides in which adjacent cysteines at positions 9 and 10 (C9, C10) are replaced by a single sarcosine residue. The peptides are shown in Table 2. Water solubility assays were performed for each peptide and the results are shown in Table 3. Substitution of sarcosines at positions A2, L3, I4, L5, I8, C9 and C9,10 resulted in peptides soluble in water. Due to their enhanced solubility in the absence of PEGylation, these peptides were selected for further evaluation of various biological activities.

[0098] Table 3. Results of Peptide Aqueous Solubility Assays

[0099]

Embodiment 2

[0100] Example 2: Complement Inhibition Assay of Peptide Variants

[0101] An assessment of the extent to which the peptide variants inhibit antibody-initiated complement activation was performed. The peptide variants were tested in two hemolytic assays: (i) ABO incompatibility ex vivo assay, and (ii) classical pathway CH50 type assay in factor B deprived serum.

[0102] In the ABO incompatibility hemolysis assay, purified erythrocytes from "AB+ type" donors are incubated with serum from "O" subjects containing anti-A and anti-B antibodies; the peptides tested is 1.8mM. The data is shown in Figure 1A middle. On an equimolar basis, variants A2, I4, I8 and C9 each inhibited ABO incompatibility hemolysis to a greater extent than the PA-dPEG24(PIC1) parent compound (P<0.015). The 18 variant reduced ABO hemolysis by 53% more than PA-dPEG24 (P<0.002). However, the C9,10 variant showed very low inhibition of ABO hemolysis.

[0103] A CH50-type hemolytic assay was then performe...

Embodiment 3

[0105] Example 3: Myeloperoxidase Inhibition and Binding

[0106] Inhibition of myeloperoxidase (MPO) activity was then tested in a TMB-based in vitro assay as previously described for PA-dPEG24 [7]. In this assay, the variants were tested for MPO inhibition over a range of concentrations. The data is shown in Figure 2A middle. Strong MPO inhibition was found for all variants with the exception of cysteine-free variants (C9,10). The half-maximal inhibition values ​​for each variant were calculated from the dose-response curves, and the data are shown in Figure 2B middle. There was a measurable difference in MPO inhibition, with variant 18 again showing the greatest efficacy among the different variants.

[0107] A plate-based assay was performed to test the binding of the peptide variants to immobilized MPO. Binding curves are shown in Figure 2C middle. MPO binding was identified for all variants except C9,10. Since the I8 and PA curves overlap almost completely, t...

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Abstract

A method of improving the lifespan of transfused platelets is described. The method may be useful for patients with alloimmunozation who are refractory to transfused platelets. A method of treating delayed hemolytic transfusion reaction is also described. Also described are PIC1 peptide variants with improved solubility and activity.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of and priority to U.S. Provisional Patent Application No. 62 / 806,432, filed February 15, 2019, and U.S. Provisional Patent Application No. 62 / 949,181, filed December 17, 2019, both of which are incorporated by reference in their entirety Incorporated into this article. Background technique [0003] Human astroviruses, belonging to the family of non-enveloped icosahedral RNA viruses, are endemic worldwide pathogens that cause acute gastroenteritis in human infants [1]. Unlike caliciviruses and rotaviruses, which cause severe acute disease, astrovirus gastroenteritis is noninflammatory [2]. The inactivated inflammatory response to astroviruses likely arises from the 787 amino acid residues of the astrovirus coat protein that form the astrovirus capsid. The coat protein has been found to inhibit the activation of the classical pathway of complement [3]. Complement is the innate immune...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61P37/02A61P37/06A61K38/16A61K45/06
CPCA61P37/02A61P37/06A61K38/1725A61K35/19A61K38/10A61K2300/00
Inventor 尼尔·K·克里希纳肯吉·昆尼昂
Owner 瑞尔塔控股有限公司
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