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. 2021 Jan-Dec:27:10760296211005544.
doi: 10.1177/10760296211005544.

Protamine Sulfate Neutralization Profile of Various Dosages of Bovine, Ovine and Porcine UFHs and Their Depolymerized Derivatives in Non-Human Primates

Ahmed Kouta  1 , Walter Jeske  1 , Lee Cera  2 , Azarfrooz Farshid  2 , Richard Duff  2 , Debra Hoppensteadt  1 , Jawed Fareed  1
Affiliations

Protamine Sulfate Neutralization Profile of Various Dosages of Bovine, Ovine and Porcine UFHs and Their Depolymerized Derivatives in Non-Human Primates

Ahmed Kouta et al. Clin Appl Thromb Hemost. 2021 Jan-Dec.

Abstract

Introduction: Currently used unfractionated heparins (UFHs) and low molecular weight heparins (LMWHs) are derived from porcine intestinal mucosa. However, heparins have also been manufactured from tissues of other mammalian species such as cow (Bovine) and sheep (Ovine). Protamine sulphate (PS) is an effective inhibitor of heparin and is used clinically to neutralize both LMWH and UFH. In this study, we determined the PS neutralization profile of these agents in non-human primate model using anti-Xa and anti-IIa methods.

Material and methods: UFHs obtained from bovine, ovine and porcine mucosal tissues and their respective depolymerized LMWHs were administered at both, gravimetric (0.5 mg/kg) and potency adjusted (100 U/kg) dosages regimen intravenously to individual groups of primates in cross over studies. PS was administered at a fixed dosage and the relative neutralization of these anticoagulants was measured utilizing amidolytic anti-Xa and anti-IIa methods.

Results: These studies have demonstrated that, the equi-gravimetric dosages of BMH, PMH and OMH have comparable PS neutralization profiles. At potency adjusted dosages, all UFHs were completely neutralized by PS. Although comparable, the LMWHs were not fully neutralized by PS in both the anti-Xa and anti-IIa assays. PS was more efficient in neutralizing the anti-IIa effects of LMWHs.

Conclusion: Heparins of diverse origins showed comparable neutralization profiles by PS in the amidolytic anti-Xa and anti-IIa assays.

Keywords: LMWHs; UFHs; bovine; ovine; porcine; protamine sulfate.

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Conflict of interest statement

Declaration of Conflicting Interests: The author(s) declared the following potential conflicts of interest with respect to the research, authorship, and/or publication of this article. Walter Jeske received research funding from KinMaster, Brazil, whereas Jawed Fareed received research funding from Ronnsi, China.

Figures

Figure 1.
Figure 1.
Comparative neutralization profile of various dosages of UFHs post protamine sulfate (PS) I.V. injection at a dose of 0.5 (mg/kg) as determined by various antiprotease assays in non-human primates (n = 4). Anti-Xa assay (A) and Anti-IIa assay (B). PMH, BMH and OMH at various dosages showed comparable PS neutralization profiles at all time points as determined by both assays. However, BMH at 100 U/kg wasn’t completely neutralized at 15- and 45-mins time points post PS injection as determined by both assays. The data represent the mean ± standard deviation (n = 4).
Figure 2.
Figure 2.
Comparative neutralization profile of various LMWHs post protamine sulfate (PS) I.V. injection at a dose of 0.5 (mg/kg) as determined by various antiprotease assays in non-human primates (n = 4). Anti-Xa assay (A) and Anti-IIa assay (B). All LMWHs were partially neutralized by PS by the same degree as determined by both assays. The data represent the mean ± standard deviation (n = 4).
Figure 3.
Figure 3.
Comparative AUC for protamine sulfate (PS) neutralization time curves of various UFHs post I.V. injection at a dose of 0.5 (mg/kg) as determined by various antiprotease assays in non-human primates (n = 4). Anti-Xa assay (A) and Anti-IIa assay (B). BMH at 0.5 mg/kg showed significant smaller AUC (P* < .05) post saline I.V. injection compared to others as measured by both assays. All UFHs at various dosages showed comparable AUC-% reduction as determined by both assays. The data represent the mean ± standard deviation (n = 4).
Figure 4.
Figure 4.
Comparative AUC for protamine sulfate (PS) neutralization time curves of various LMWHs post I.V. injection at a dose of 0.5 (mg/kg) as determined by various antiprotease assays in non-human primates (n = 4). Anti-Xa assay (A) and Anti-IIa assay (B). All LMWHs showed comparable AUC-% reduction as determined by both assays. The data represent the mean ± standard deviation (n = 4).
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