Efficacy of Intracolonic Administration of Low-Molecular-Weight Heparin CB-01-05, Compared to Other Low-Molecular-Weight Heparins and Unfractionated Heparin, in Experimentally Induced Colitis in Rat

Abstract

Purpose Parenteral administration of lowmolecular-weight heparins (LMWHs) and unfractionated heparin (UFH) resulted effective in improving the symptoms of experimental colitis in rat. Today, there is little information about their activity by intracolonic instillation. The scope of this study was to evaluate the ability of CB-01-05 (a LMWH with a mean molecular weight of about 5,700), compared to a series of other LMWHs and to UFH, directly instilled into the distal colon of the rat, to ameliorate dinitrobenzene (DNB)-induced experimental colitis. Method Adult male Wistar rats underwent colitis induction by intracolonic instillation of DNB. Starting 24 h after colitis induction, CB-01-05 (0.005-0.9 mg), other LMWHs (0.3-0.6 mg), and UFH (0.6 mg) were instilled, by rectal route, into the distal colon once a day for three consecutive days. On the day following the last administration, the animals were sacrificed and the distal colon was isolated, weighed, macroscopically examined, and processed for histology. Additional experiments in rat splenocytes, performed in order to elucidate the anti-inflammatory mechanisms of CB-01-05, were performed. Results Among the tested items, only CB-01-05 at doses ranging from 0.2 to 0.9 mg was significantly effective in reducing colon weight increase and in improving both the mucosal damaged area and the histological score. The other LMWHs resulted far less effective, showing decreasing activity closely related with the decrease of their molecular weight, thus demonstrating their biological nonequivalence. CB-01-05 resulted also more active than UFH. CB-01-05 was shown to interfere with cytokines production by rat splenocytes, mainly inhibiting interferon (IFN)-c expression. Conclusions CB-01-05 instilled into the colon is well tolerated, has strong anti-inflammatory effect on DNBinduced colitis in rat, and is the most effective agent among other LMWHs and UFH. These results suggest that the anti-inflammatory activity of CB-01-05, together with its topical administration, could represent a new approach in the management of ulcerative colitis.

Clinical trial: oral colon‐release parnaparin sodium tablets (CB‐01‐05 MMX®) for active left‐sided ulcerative colitis

“…Previous in vivo 10 and in vitro 14 studies, and a pilot human trial 11 suggested a potential anti‐inflammatory effect of parnaparin sodium in the colon.…”

“…In spite of evidence that LMWHs have been shown to be effective by oral administration, 9 this route has been considered inappropriate to treat UC because of the conviction that the LMWHs are quickly inactivated in the upper gastrointestinal tract therefore not reaching adequate concentrations in the colonic lumen. Recent observations proved that parnaparin sodium, an LMWH with a mean molecular mass around 5000, delivered by catheter into the colon of rat was highly effective in ameliorating dinitrobenzene (DNB)‐induced colitis 10 . This observation suggested that the administration of parnaparin sodium, contained in tablets delivering the product directly into the lumen of the colon, could represent a promising approach to treat UC.…”

Calcium butyrate: Anti-inflammatory effect on experimental colitis in rats and antitumor properties

“…Groups of 8-10 male Wistar rats (Charles River, Lecco, Italy) for a total of 27 animals weighing ~200-250 g were used. The test was performed as previously described (31), with the exception of a longer treatment duration. Colitis was induced in animals fasted overnight and under light ether anaesthesia, by instillation into the colonic lumen of 30 mg DNB dissolved in 0.25 ml ethanol 50%.…”

Central Role of the Gut Epithelial Barrier in the Pathogenesis of Chronic Intestinal Inflammation: Lessons Learned from Animal Models and Human Genetics

“…Data recently obtained using chemically induced colitis models suggest a potential role of two different LMWHs in ameliorating gut inflammation if administered directly on the epithelial surface. In one study, parnaparin, a LMWH of 5000 kD, was administered to rats intracolonically, and improved disease severity (156). In another study, colitic mice treated with microspheres loaded with enoxaparin (4500 kD), specifically designed to release the drug into the colon, resulted in a reduction of inflammatory activity after parenteral administration (157).…”