Inflammatory activity in vivo

Researchers (Nuijens et al., 1997) at the Leiden University (the Netherlands) in collaboration with Pharming, NV (Leiden, the Netherlands) compared recombinant human lactoferrin (rhLF) expressed in the milk of transgenic mice with natural human milk-derived lactoferrin (hLF). They concluded that the unsaturated rhLF and natural hLF had comparable properties, indicating that hLF produced in bovine milk will exert similar, if not identical, antibacterial and anti-inflammatory activities in vivo. Pharming also developed the first transgenic bull in the late... 

Amentoflavone (1), a biflavone also isolated from Selaginella species, showed potent anti-inflammatory activity in vivo, by i.p. route had approximately 1/2-1/5 of the anti-inflammatory activity of indomethacin or prednisolone against several animal models of acute inflammation, including acetic acid-induced writhings in mice. However, amentoflavone (1) did not significantly reduce adjuvant-induced arthritis (AIA) in rats.[31]... 

Ungar, Damgaard and Weinstein have shown that certain well recognized anti-inflammatory drugs, salicylic acid, phenazone, amidopyrine, cinchophen and also jf)-aminophenol inhibit fibrinolysin (plasmin) in concentrations close to those which show anti-inflammatory activity in vivo. Salicylate may inhibit kinin formation by this indirect mechanism involving plasmin. The failure of some workers to observe any inhibition of kallikrein by salicylate in vitro does not conflict with this. 

ES-62 is a large and hence immunogenic molecule whose posttranslational addition of PC is dependent on a nematode-spedfic pathway. For these reasons, it is not suitable for use as drug. However, small PC-based derivatives oflFer a viable alternative. Towards this end we have shown that small PC-containii molecules (e.g., PC- cans) can mimic some of the activities of ES-62 in vitro. The next step will be to demonstrate that the same is true with respect to ES-62 s anti-inflammatory activity in vivo. 

Guilford W. J., Bauman J. G., SkubaUa W., Bauer S., Wei G. P, Davey D., Schaefer C. et al. 2004. Novel 3-oxa lipoxin A4 analogues with enhanced chemical and metabolic stability have anti-inflammatory activity in vivo. 

Rajan S G et al. [74] designed and S3mthesized a sequence of 2-(2,4-disubstituted-thiazole-5-yl)-3-aryl-3H-quinazoline-4-one 23 compounds. S3mthesized molecules were estimated for their inhibitory activity in the course of record factors, nuclear factor-kB (NF-kB) and activating factor (AP-1) interceded transcriptional activation in a cell line based in vitro assay as well as for their anti-inflammatory activity in vivo model of severe inflammation. 

Bristol-Myers Squibb has recently disclosed two different series of carbamate-based FAAH inhibitors. The first of these is a series of 4,5-diaryl-imidazoles in which 30 compounds are specifically claimed, an example being compound (57). This compound is reported to have an IC50 value of < 10 nM. In addition, (57) was also active in vivo in rodent models of chemo-induced, thermal and neuropathic pain [72]. The second series of compounds is based on oxime carbamoyl FAAH inhibitors such as (58). Compound (58) is reported to have an IC50 value of < 10 nM and activity in rodent models of inflammatory pain, thermal pain and inflammatory oedema [73]. 

In other cases, the widespread application of a biopharmaceutical may be hindered by the occurrence of relatively toxic side effects (as is the case with tumour necrosis factor a (TNF-a, Chapter 9). Finally, some biomolecules have been discovered and purified because of a characteristic biological activity that, subsequently, was found not to be the molecule s primary biological activity. TNF-a again serves as an example. It was first noted because of its cytotoxic effects on some cancer cell types in vitro. Subsequently, trials assessing its therapeutic application in cancer proved disappointing due not only to its toxic side effects, but also to its moderate, at best, cytotoxic effect on many cancer cell types in vivo. TNF s major biological activity in vivo is now known to be as a regulator of the inflammatory response. 

Go 6850 (GF-109203X) (485), a dimethylaminoethyl analog of 2,3-bis(indol-3-yl) maleimide was shown to be a selective inhibitor of PKC with anti-inflammatory activity in several in vivo and in vitro murine models of acute inflammation (432) (Scheme 4.7). 

Rodent KC and HC, as well as human HC, express an inducible NO synthase under septic or inflammatory conditions. In vivo in endotoxemia, this expression is transient. Our in vivo data indicate that this induced -NO serves a protective role in the liver and reduces hepatic injury in endotoxemia. This protective action may be mediated by the capacity of NO to neutralize oxygen radicals and prevent platelet adherence and aggregation. Our in vitro studies show that HC-derived -NO can activate soluble guanylate cyclase. Other in vitro effects include the nonspecific suppression of protein synthesis and a small reduction in mitochondrial aconitase activity. The relevance of these in vitro actions to hepatic function in vivo remains to be determined. 

Five lignans isolated from the heartwood of T. baccata L. growing in Turkey were evaluated for their anti-inflammatory and antinociceptive activities in vivo. All of the compounds were shown to possess significant antinociceptive activities against p-bcnzoquinone-induccd abdominal contractions and significantly inhibited carrageenan-induced hind paw edema in mice [100]. 

Azo Prodrugs Amines have been incorporated into an azo linkage to form prodrugs that can be activated through azo reduction. In fact, sulfa dmgs were discovered because of prontosil (93), an inactive azo dye that was converted in vivo to the active sulfanilamide (95) (Scheme 15). Clinically useful balsalazide (23), olsalazine (25), and sulfasalazine (26) are azo prodrugs of mesalazine (27). They are converted in vivo by bacterial azo reductases in the gut to the active 5-aminosalicylic acid (5-ASA or mesalazine, 27), which is responsible for their anti-inflammatory activity in the treatment of ulcerative colitis, as discussed earlier. 

The 2-ainino-l-(i3-D-ribofuranosyl)ethanol derivative 702 was condensed with 2,3-dichIoropyrazine to give 703, which was elaborated to 707 [93JHC(31)1213] (Scheme 180). C-Nucleoside 707 demonstrated weak suppression of mouse splenic NK-cell activity toward YAC lymphoma cells and in vivo anti-inflammatory activity in rats [93JHC(31)1213]. 

Whilst it seems possible that certain of the effects of anti-inflammatory compounds in vivo may be explained in terms of uncoupling activity (e.g. the increased oxygen consumption and hyperthermia in animals and man subjected to overdoses of acetylsalicylic acid), further work is needed to establish that lower doses of the drugs do, in fact, cause uncoupling, and that their anti-inflammatory activity is dependent on this. 

Zebrafish have emerged as a powerfiil model organism to study neutrophil chemotaxis and inflammation in vivo. Studies of neutrophil chemotaxis in animal models have previously been hampered both by the limited number of specimens available for analysis and by the need for invasive procedures to perform intravital microscopy. Due to the transparency and cell permeability of zebrafish embryos these limitations are circumvented, and the zebrafish system is amenable to both live time-lapse imaging of neutrophil chemotaxis and for screening of the effects of chemical compounds on the inflammatory response in vivo. Here, we describe methods to analyze neutrophil-directed migration toward wounds using both fixed embryos by myeloperoxidase activity assay, and live embryos by time-lapse microscopy. Further, methods are described for the evaluation of the effects of chemical compounds on neutrophil motility and the innate immune responses in zebrafish embryos. 

---