Natural polymer/drug carrier; Biochemical reagent

At lower concentrations of 40 μg/mL, κ-Car-Curcumin (Cur) (0-500 μg/mL; 24-72 hours) significantly inhibits the development of cancer cells[1]. In certain A549 lung cancer cells, the cytotoxicity of the Cur-loaded κ-Car dramatically increases apoptotic activity[1]. In HT-29 cells, κ-carrageenan (1–60 μg/mL; 0.5–24 hours) increases LPS-induced IL-8 secretion[2].

κ-Carrageenan can be applied to animal modeling to create models of edema in the feet of rats and mice.

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Background. The dietary usage of carrageenan as common food additive has increased observably over the last 50 years. But there is substantial controversy about its safety. Methods. We investigated whether the κ-carrageenan could enhance lipopolysaccharide-induced IL-8 expression by studying its actions on the TLR4-NF-κB pathway. The aggravating effect of κ-carrageenan on Citrobacter freundii DBS100-induced intestinal inflammation was also investigated in a mouse model. Results. Our data show that κ-carrageenan pretreatment promoted LPS-induced IL-8 expression in HT-29 cells. Although CD14, MD-2, and TLR4 were upregulated, the binding of LPS was not enhanced. However, the pathway of Bcl10-NF-κB was triggered. Interestingly, κ-carrageenan competitively blocked the binding of FITC-LPS. Furthermore, pretreatment with κ-carrageenan for one week previous to gavage with C. freundii DBS100 markedly aggravated weight loss, mortality, and colonic damage. The secretion of cytokines was unbalanced and the ratio of Tregs was decreased significantly. In addition, κ-carrageenan, together with C. freundii DBS100, enhanced the transcription and secretion of TLR4 and NF-κB. Conclusions. κ-Carrageenan can synergistically activate LPS-induced inflammatory through the Bcl10-NF-κB pathway, as indicated by its aggravation of C. freundii DBS100-induced colitis in mice. General Significance. Our results suggest that κ-carrageenan serves as a potential inflammatory agent that magnifies existing intestinal inflammation[2].

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In the present study, we investigated the effects of κ-carrageenan on TNBS-induced gut inflammation in mice. BALB/c mice were pretreated with κ-carrageenan for 14days prior to the administration of TNBS. Results: Our results showed that κ-carrageenan pretreatment aggravated the loss of body weight and further increased the mortality rate. Histological and morphological analyses revealed that the TNBS-induced colonic inflammation was deteriorated by the κ-carrageenan administration. The ratio of CD4(+)CD25(+)CD127dim/CD4(+) of the κ-carrageenan+TNBS groups was significantly lower than that of the TNBS group. The expression of IL-2, TNF-α and IL-6 was significantly increased, whereas the expression of IL-10 was significantly decreased in the κ-carrageenan+TNBS groups. In addition, κ-carrageenan, together with TNBS, decreased the enzyme activity of SOD and GSH-px and up-regulated the expression of TLR4, NF-κB, p-ERK, p-JNK, p-Jun., IL-8 and MDA in the colonic mucosa. Conclusions: κ-Carrageenan aggravated the TNBS-induced intestinal inflammation, and such an effect could be associated with the oxidative stress and activation of TLR4-NF-κB and MAPK/ERK1/2 pathway[3].

The current study is to develop a natural drug carrier with seaweed derived polymers namely κ-Carrageenan (κ-Car) for drug delivery applications. κ-Car is a natural polysaccharide which derived from edible red seaweeds, they are easily available, non-toxic, cost effective, biodegradable and biocompatible nature. Curcumin (Cur) is a yellow-orange polyphenol existing in turmeric, which is predominantly used as spice and food coloring agent. The ultimate use of polymeric composites, especially those composed of natural polymers, has become a very interesting approach in recent drug delivery applications, due to their non-toxicity and biological origin. In this study the primary approach which depends on the loading of Curcumin into κ-Carrageenan was accomplished, and which (κ-Car-Cur) an active drug carrier was developed for drug delivery against selected lung cancer cells (A549). Thus, the κ-Car-Cur was synthesized by solvent evaporation method followed by freeze drying, and it was further characterized. From this study, it has been reported that the high encapsulation efficiency, good stability, and successful release of Cur from the carrier (κ-Car) was achieved. The drug release was more active at acidic pH 5.0 with the cumulative release of 78%, which is the favorable condition present in tumor microenvironments. The in vitro cellular applications studies of κ-Car-Cur demonstrated that, κ-Car-Cur composites induced higher cytotoxicity against selected cancer cells than free Cur and effectively involved to trigger cellular apoptosis in A549 cancer cells. Further, it was also possessed that inhibition of cell growth and changes in metabolic activity of cancer cells are the unique characteristic features of cellular apoptosis, through reactive oxygen species (ROS) generation. It also observed that there was a decrease in mitochondrial membrane potential (ΔψmΔψm) which leads to a cellular apoptosis during treatment with κ-Car-Cur. Hence, the study outcomes may provide the potential outline for the use of κ-Car-Cur as a promising tool to deliver drugs at intracellular level[1].

Cell Viability Assay[1]
Cell Types: A549 cells
Tested Concentrations: 0-500 μg/mL
Incubation Duration: 24, 48 and 72 hrs (hours)
Experimental Results: The dose response effects of cells treated with Cur loaded κ-Car after incubation of 24, 48 and 72 h demonstrated significant IC50 values of 65, 50 and 40 μg/mL respectively, for 24, 48, 72 h ours.

Animal/Disease Models: Male and female NIH (s) mice[2]
Doses: 1.7 mg/kg, LOW; 8.3 mg/kg, MED; or 41.7 mg/kg, HIG
Route of Administration: Orally administered for 1 week prior to C. freundii DBS100 treatment
Experimental Results: Enhanced the C. freundii DBS100-dependent induction of TLR4 and NF-κB in the intestinal mucosa of infected mice.

[1]. κ-Carrageenan: An effective drug carrier to deliver curcumin in cancer cells and to induce apoptosis. Carbohydr Polym. 2017;160:184-193.

[2]. κ-Carrageenan Enhances Lipopolysaccharide-Induced Interleukin-8 Secretion by Stimulating the Bcl10-NF-κB Pathway in HT-29 Cells and Aggravates C. freundii-Induced Inflammation in Mice. Mediators Inflamm. 2017;2017:8634865.

[3]. Enhanced effect of κ-carrageenan on TNBS-induced inflammation in mice. Int Immunopharmacol. 2016;39:218-228.

In conclusion, we demonstrated enhanced effects of κ-carrageenan on the inflammatory reaction in vitro and in vivo. κ-Carrageenan increased the expression of TLR4 receptor but competitively bound TLR4, blocking LPS binding, which rules out the enhanced TLR4 levels as an explanation for the synergistic effects of κ-carrageenan. However, our results also showed that κ-carrageenan can participate in the Bcl10-NF-κB-mediated pathway to enhance LPS stimulated secretion of IL-8 in HT-29 cells. The C. freundii DBS100-induced intestinal inflammation model further verified that κ-carrageenan could aggravate the inflammatory reaction of the colon to pathogen exposure κ-carrageenan modulated cytokine production, downregulated the proportion of Tregs, and upregulated NF-κB, all of which are likely to contribute to the exacerbatory effects of κ-carrageenan. Collectively, our results suggest that κ-carrageenan serves as a potential inflammatory agent that magnifies existing intestinal inflammation.[2]

788.099

11114-20-8

11966249

Off-white to light yellow solid powder

-8.5

8

25

10

51

1380

20

S(=O)(=O)([O-])O[C@@]1([H])[C@@]([H])(C([H])([H])O[H])O[C@]([H])([C@@]([H])([C@@]1([H])O[C@]1([H])[C@@]([H])([C@]2([H])[C@]([H])([C@@]([H])(C([H])([H])O2)O1)O[C@@]1([H])[C@@]([H])([C@]([H])([C@]([H])([C@@]([H])(C([H])([H])O[H])O1)OS(=O)(=O)[O-])O[H])O[H])O[H])O[H])O[C@@]1([H])[C@@]2([H])C([H])([H])O[C@]1([H])[C@]([H])([C@@]([H])(O[H])O2)O[H]

ZNOZWUKQPJXOIG-XSBHQQIPSA-L

InChI=1S/C24H38O25S2/c25-1-5-14(48-50(33,34)35)9(27)10(28)22(42-5)45-16-8-4-40-19(16)12(30)23(44-8)47-20-13(31)24(43-6(2-26)17(20)49-51(36,37)38)46-15-7-3-39-18(15)11(29)21(32)41-7/h5-32H,1-4H2,(H,33,34,35)(H,36,37,38)/p-2/t5-,6-,7-,8-,9-,10-,11-,12-,13-,14+,15+,16+,17+,18-,19-,20-,21+,22+,23-,24+/m1/s1

[(2R,3S,4R,5R,6S)-6-[[(1R,3S,4R,5R,8S)-3,4-dihydroxy-2,6-dioxabicyclo[3.2.1]octan-8-yl]oxy]-4-[[(1R,3R,4R,5R,8S)-8-[(2S,3R,4R,5R,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-sulfonatooxyoxan-2-yl]oxy-4-hydroxy-2,6-dioxabicyclo[3.2.1]octan-3-yl]oxy]-5-hydroxy-2-(hydroxymethyl)oxan-3-yl] sulfate

11114-20-8; [(2R,3S,4R,5R,6S)-6-[[(1R,3S,4R,5R,8S)-3,4-dihydroxy-2,6-dioxabicyclo[3.2.1]octan-8-yl]oxy]-4-[[(1R,3R,4R,5R,8S)-8-[(2S,3R,4R,5R,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-sulfonatooxyoxan-2-yl]oxy-4-hydroxy-2,6-dioxabicyclo[3.2.1]octan-3-yl]oxy]-5-hydroxy-2-(hydroxymethyl)oxan-3-yl] sulfate; Vegetable gelatin; ZNOZWUKQPJXOIG-XSBHQQIPSA-L; YC30039

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O: 8.33 mg/mL
DMSO: 8.33 mg/mL

Solubility in Formulation 1: ≥ 0.83 mg/mL (Infinity mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 8.3 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 0.83 mg/mL (Infinity mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 8.3 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 0.83 mg/mL (Infinity mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 8.3 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 8.33 mg/mL (Infinity mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication (<60°C).

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 (Please use freshly prepared in vivo formulations for optimal results.)