Low molecular weight protamine acetate (LMWP acetate;TDSP5 acetate) is a truncated arginine-rich protamine peptide, which also acts as an antidote for heparin/low molecular weight heparin and a cell-penetrating delivery vector. Low molecular weight protamine acetate neutralizes heparin-induced anticoagulant activities, including aPTT, anti- Xa and anti- IIa activities. Low molecular weight protamine acetate translocates across mammalian cell membranes, delivers conjugated impermeable molecules through tumor tissues, enhances the skin permeability of conjugated epidermal growth factor, and accelerates wound healing when conjugated with epidermal growth factor. Low molecular weight protamine acetate retains the in vitro cell proliferation activity of conjugated EGF, and also enables site-specific conjugation with peptides or proteins via genetic recombination. Low molecular weight protamine acetate can be used in studies related to colon cancer, skin wounds and diabetic skin wounds [1] [2] [3].
In Vivo, Low molecular weight protamine-gelonin conjugate (110 μg; peritumoral injection; nine times over 15 days) acetate achieves significant colon adenocarcinoma tumor regression, reducing average tumor weight to 0.33 g in BALB/c mice[2].
In Vitro, Low molecular weight protamine (18 μg/U; 3 min) acetate completely neutralizes the aPTT activity of 5 U/mL heparin in human plasma at a ratio of 18 μg/U, requiring a dose ~2.4-fold higher than protamine[1]. Low molecular weight protamine (12 μg/U; 10 min) acetate completely neutralizes the anti-IIa activity of 1 U/mL heparin in human plasma at a ratio of 12 μg/U, requiring a dose ~1.6-fold higher than protamine[1]. Low molecular weight protamine (30 μg/μg) acetate completely neutralizes the anti-Xa activity of Mono-Embolex at a ratio of 30 μg/μg, with efficacy similar to protamine[1]. Low molecular weight protamine (100 μg/U) acetate completely neutralizes the anti-Xa activity of both Fragmin and LMWH3000 at a ratio of 100 μg/U, demonstrating broad efficacy against low molecular weight heparins[1]. Low molecular weight protamine (4.5 mg/mL; 60 min at 37°C) acetate induces only 30% complement consumption in diluted human serum, showing drastically reduced complement activation potential compared to protamine[1]. Heparin-Low molecular weight protamine acetate complexes (22 μg/U; 60 min at 37°C), formed at a ratio of 22 μg/U, induce only ~50% complement consumption in human serum with 2.5-10 U/mL heparin, showing reduced complement activation potential compared to heparin-protamine complexes[1]. Low molecular weight protamine (63 μM; 2 h at 37°C) acetate acetate cross-reacts with mouse antiprotamine antibodies with a IC50 of 63 μM, showing ~5.25-fold lower cross-reactivity than protamine[1]. The Low molecular weight protamine acetate-gelonin conjugate potently inhibits protein synthesis in a cell-free rabbit reticulocyte lysate system with an IC50 of ~23 pM[2]. FITC-labeled Low molecular weight protamine (TDSP5) (0.2 μM; 30 min) acetate is efficiently taken up by 293T, HeLa, CT-26, MCF-7, NIH3T3, and MG63 cells, with uptake efficiency equivalent to TAT peptide and protamine and unaffected by 10% serum[2]. Low molecular weight protamine (TDSP5) acetate (0.2 μM; 30 min) efficiently translocates into HeLa, CT-26, and MG63 cells, localizes to the cytoplasm, and effectively delivers conjugated phalloidin and gelonin into cells[2]. Low molecular weight protamine (TDSP5) (0.01-10.0 mM; 72 h) acetate exhibits minimal cytotoxicity toward 293T cells, with less than 10% reduction in cell viability at concentrations up to 10.0 mM after 72 h incubation[2]. The Low molecular weight protamine acetate -gelonin conjugate (1:1 molar ratio) (48 h) potently inhibits CT-26 colon adenocarcinoma cell growth with an IC50 of 39 pM after 48 h incubation, while free gelonin or free LMWP plus gelonin have no effect[2].
References:
[1]. Chang LC, et al. Low molecular weight protamine (LMWP) as nontoxic heparin/low molecular weight heparin antidote (II): in vitro evaluation of efficacy and toxicity. AAPS PharmSci. 2001;3(3):E18.
[2]. Park YJ, et al. Nontoxic membrane translocation peptide from protamine, low molecular weight protamine (LMWP), for enhanced intracellular protein delivery: in vitro and in vivo study. FASEB J. 2005;19(11):1555-1557.
[3]. Choi JK, et al. The effect of epidermal growth factor (EGF) conjugated with low-molecular-weight protamine (LMWP) on wound healing of the skin. Biomaterials. 2012;33(33):8579-8590.
In Vivo, Low molecular weight protamine-gelonin conjugate (110 μg; peritumoral injection; nine times over 15 days) acetate achieves significant colon adenocarcinoma tumor regression, reducing average tumor weight to 0.33 g in BALB/c mice[2].
In Vitro, Low molecular weight protamine (18 μg/U; 3 min) acetate completely neutralizes the aPTT activity of 5 U/mL heparin in human plasma at a ratio of 18 μg/U, requiring a dose ~2.4-fold higher than protamine[1]. Low molecular weight protamine (12 μg/U; 10 min) acetate completely neutralizes the anti-IIa activity of 1 U/mL heparin in human plasma at a ratio of 12 μg/U, requiring a dose ~1.6-fold higher than protamine[1]. Low molecular weight protamine (30 μg/μg) acetate completely neutralizes the anti-Xa activity of Mono-Embolex at a ratio of 30 μg/μg, with efficacy similar to protamine[1]. Low molecular weight protamine (100 μg/U) acetate completely neutralizes the anti-Xa activity of both Fragmin and LMWH3000 at a ratio of 100 μg/U, demonstrating broad efficacy against low molecular weight heparins[1]. Low molecular weight protamine (4.5 mg/mL; 60 min at 37°C) acetate induces only 30% complement consumption in diluted human serum, showing drastically reduced complement activation potential compared to protamine[1]. Heparin-Low molecular weight protamine acetate complexes (22 μg/U; 60 min at 37°C), formed at a ratio of 22 μg/U, induce only ~50% complement consumption in human serum with 2.5-10 U/mL heparin, showing reduced complement activation potential compared to heparin-protamine complexes[1]. Low molecular weight protamine (63 μM; 2 h at 37°C) acetate acetate cross-reacts with mouse antiprotamine antibodies with a IC50 of 63 μM, showing ~5.25-fold lower cross-reactivity than protamine[1]. The Low molecular weight protamine acetate-gelonin conjugate potently inhibits protein synthesis in a cell-free rabbit reticulocyte lysate system with an IC50 of ~23 pM[2]. FITC-labeled Low molecular weight protamine (TDSP5) (0.2 μM; 30 min) acetate is efficiently taken up by 293T, HeLa, CT-26, MCF-7, NIH3T3, and MG63 cells, with uptake efficiency equivalent to TAT peptide and protamine and unaffected by 10% serum[2]. Low molecular weight protamine (TDSP5) acetate (0.2 μM; 30 min) efficiently translocates into HeLa, CT-26, and MG63 cells, localizes to the cytoplasm, and effectively delivers conjugated phalloidin and gelonin into cells[2]. Low molecular weight protamine (TDSP5) (0.01-10.0 mM; 72 h) acetate exhibits minimal cytotoxicity toward 293T cells, with less than 10% reduction in cell viability at concentrations up to 10.0 mM after 72 h incubation[2]. The Low molecular weight protamine acetate -gelonin conjugate (1:1 molar ratio) (48 h) potently inhibits CT-26 colon adenocarcinoma cell growth with an IC50 of 39 pM after 48 h incubation, while free gelonin or free LMWP plus gelonin have no effect[2].
References:
[1]. Chang LC, et al. Low molecular weight protamine (LMWP) as nontoxic heparin/low molecular weight heparin antidote (II): in vitro evaluation of efficacy and toxicity. AAPS PharmSci. 2001;3(3):E18.
[2]. Park YJ, et al. Nontoxic membrane translocation peptide from protamine, low molecular weight protamine (LMWP), for enhanced intracellular protein delivery: in vitro and in vivo study. FASEB J. 2005;19(11):1555-1557.
[3]. Choi JK, et al. The effect of epidermal growth factor (EGF) conjugated with low-molecular-weight protamine (LMWP) on wound healing of the skin. Biomaterials. 2012;33(33):8579-8590.