**6.3 Microbial and plant derived lipases**

With the aim of developing porcine-free enzyme supplements and in order to avoid the short-life of lipolytic enzymes of pancreatic origin, microbial lipases of fungal or bacterial origin were suggested for replacement therapy. Therefore, potential efficacy of many fungal Lipases derived from *Aspergillus niger* (Griffin & al., 1989), *Rhizopus arrhizus* (Iliano & Lodewijk, 1990), *Rhizopus delemar* (Galle & al., 2004 ), *Candida cylindracea* (Schuler & Schuler, 2008) and *Yarrowia lipolytica* (Turki & al., 2010a) was investigated. The *Yarrowia lipolytica* lipase seemed to be of potential interest because of its acid and protease-stable properties and its resistance to the detergent action of bile salts as shown *in vitro* (Turki & al., 2010a). Supporting its use as a pharmaceutical, safety assessment of the enzyme in rats showed that there were no toxicologically severe changes in clinical signs, growth, hematology, clinical chemistry, organ weight and pathology related to oral administration of *Yarrowia lipolytica* lipase in animals (Turki & al., 2010b). Actually, a substitute for EPI treatment based on Lip2p is under investigation by Laboratoire Mayoly Spindler, a French pharmaceutical company specialized in gastroenterology therapeutics (Fickers & al., 2011, as cited in http://www.mayoly-spindler.com/). The process development in cGMP conditions for the production of the *Yarrowia lipolytica* MS1819 lipase was completed in 2009 with the Swiss biotech company DSM Nutritional Product Ltd (Fickers & al., 2011, as cited in http://www.dsm.com, press release, December 22th, 2009). In 2010, a drug development partnership was established with Protea Biosciences to initiate phase I/IIA clinical trials in France with the aim of demonstrating safety and proof-of-concept of the therapeutic use of this recombinant lipase (Fickers & al., 2011).

A pipeline preparation Liprotamase (formerly known as ALTU 135 and Trizytek) containing bacterial lipase, fungal protease and amylase was developed by Eli Lilly company (Eli Lilly, IN, USA, www. Lilly.com). An open-label Phase III safety study was carried out in order to evaluate 214 patients, of which 145 CF patients, ages 7 and above, completed 12 months of treatment with Liprotamase. Investigators found that 96 percent of all CF patients who received liprotamase for 12 months maintained or gained weight. Based on key nutritional parameters, the study showed that patients who completed 12 months of treatment with

Emerging Approaches for the Treatment of

remains yet to be assessed.

**8. References** 

Fat Malabsorption due to Exocrine Pancreatic Insufficiency 287

of nutrients by the intestines may result, leading to deficiencies of essential nutrients and the occurrence of loose stools containing unabsorbed fat (steatorrhea). A shortage of the digestive enzymes necessary to break down food is the main cause of this dietary malabsorption. Unlike protein and starch digestion, lipid malabsorption is the overriding problem and the main cause of clinical symptoms and nutritional deficiencies. Until recently, approaches used to address problem of fat malabsorption due to pancreatic insufficiency have been focusing primarily on oral administration of exogenous pancreatic enzymes extracted from porcine source. Standard clinical practices dictate administration of lipase 25,000-75,000 units/meal by using pH-sensitive pancrelipase microspheres, along with dosage increases, compliance checks, and differential diagnosis in cases of treatment failure. Various pancreatin preparations are available, however, differences in galenic properties and release kinetics and other factors such as early acid inactivation, under dosage and patient incompliance may decrease clinical efficacy of the treatment. The FDA decreed that all manufacturers of pancreatic enzyme supplements must file new drug applications (NDA) to ensure consistent efficacy, safety, and quality of these agents. Accordingly, improved approaches to treat efficiently problem of fat malabsorption secondary to pancreatic insufficiency are investigated. New alternatives of enzyme substitution therapy are being developed. Emerging therapeutic landscape includes use of porcine free - lipase preparations. Enzyme supplements either from human, mammalian, microbial or plant origins are wisely suggested. Interestingly, newest approaches state the design of acid -stable variants of human pancreatic lipase as well as creation of functional dietary food with specific more digestible/absorbable lipid sources. However, how these pipeline therapies may help meet the ongoing challenges in treating lipid malabsorption in patients with pancreatic insufficiency and improve the long-term outcomes of these patients

Abdelkafi, S.; Fouquet, B. ; Barouh, N. ; Durner, S. ; Pina, M. ; Scheirlinckx, F. ; Villeneuve, P.

Aloulou, A. ; Puccinelli, D. ; Sarles, J. ; Laugier, R. ; Leblond, Y. & Carrière, F. (2008). In vitro

Ameis, D.; Stahnke, G.; Kobayashi, J.; McLean, J.; Lee, G.; Buscher, M.; Schotz, M.C. & Will,

Armand, M.; Borel, P.; Dubois, C.; Senft, M.; Peyrot, J.; Salducci, J.; Lafont, H. & Lairon, D.

Armand, M.; Hamosh, M.; Mehta, N.R.; Angelus, P.A.; Philpott, J.R.; Henderson, T.R.;

therapy. *Food Chemistry*, Vol.115, No. 2, pp. 488–494

*Biological Chemistry.* Vol. 265, pp. 6552–6555.

Vol. 27, No. 3, pp. 283-392.

266, No. 3, pp. 372-381.

& Carrière F. (2009). In vitro comparisons between Carica papaya and pancreatic lipases during test meal lipolysis: Potential use of CPL in enzyme replacement

comparative study of three pancreatic enzyme preparations: dissolution profiles, active enzyme release and acid stability. *Alimentary Pharmacology and Therapeutics.*

H. (1990). Isolation and characterization of the human hepatic lipase gene. *Journal of* 

(1994).Characterization of emulsions and lipolysis of dietary lipids in the human stomach. *American Journal of Physiology Gastrointestinal and Liver Physiology*, Vol.,

Dwyer, N.K.; Lairon, D. & Hamosh, P. (1996). Effect of human milk or formula on

liprotamase demonstrated that they maintained their nutritional status; and survival in people living with cystic fibrosis was maintained too (Borowitz & al., 2011). Subsequent to the completion of the stage III clinical study on liprotamase, the drug's manufacturer submitted a New Drug Application to the U.S. Food and Drug Administration (FDA) for approval. However, on January 13th of this year, the FDA panel stated that he was not convinced that Liprotamase was any better than the current pancreatic enzyme products available now. The manufacturer disclosed that another clinical trial must be conducted before the FDA will consider the approval of this drug (Eli Lilly, IN, USA, www. Lilly.com).

In yet other approaches, plant acid-stable lipases were suggested as good alternatives to porcine preparations. Hence, considerable attention has focused in these enzymes and suitable techniques for isolating and purifying them have been well documented. A lipase sourced from *Carica papaya* latex has been recently proposed as suitable candidate for use as a therapeutic tool in patients with pancreatic exocrine insufficiency (Abdelkafi et al., 2009). The enzyme showed several biochemical properties enabling it to act in the gastro-intestinal tract like mammalian digestive lipases (Abdelkafi et al., 2009): (i) its activity on long-chain Triacylglycerols reaches an optimum at pH 6.0 in the presence of bile, (ii) it is only weakly inhibited by bile salts, (iii) it shows a similar pattern of regioselectivity to that of human pancreatic lipase, generating 2-Mono acylglycerol and free fatty acids (FFA), the lipolysis products absorbed at the intestinal level, and (iv) it shows significant levels of stability and activity at low pH values at a temperature of 37 °C. Therefore, *Carica papaya* lipase seems to be tailored to act optimally under the physiological conditions pertaining in the gastrointestinal tract. However, its sensitivity to digestive proteases still needs to be tested.
