Razvoj in vrednotenje formulacij za peroralno uporabo laktoferina, izdelanih z mikrokapsuliranjem in filmskim oblaganjem

Brečko, Katja (author), Gašperlin, Mirjana (mentor)

Laktoferin je glikoprotein, ki ga izločajo številne žleze v telesu in ga najdemo tudi v mleku. Široka zastopanost v telesu govori o velikem pomenu laktoferina za organizem. Njegovi zaščitni učinki segajo od protimikrobne aktivnosti do protivirusnega in protirakavega delovanja. Tako širok spekter aktivnosti omogočajo mehanizmi delovanja laktoferina, ki izkazujejo njegovo sposobnost vezave železa in interakcije z molekularnimi in celičnimi komponentami gostitelja in patogenov. Zaradi številnih koristi laktoferina in z namenom zmanjšanja okoljskega bremena, ki ga povzroča odpadna kisla sirotka, smo izdelali prehransko dopolnilo v obliki mikrosfer. Za izdelavo mikrosfer smo uporabili metodo kapljanja raztopine Na-alginata s pomočjo peristaltične črpalke v raztopino Ca2+ ionov. Z napravo za mikrokapsuliranje Inotech IE-50R smo izdelovali mikrosfere manjših velikosti. Majhna količina laktoferina se vgradi v alginatne mikrosfere, zato smo z absorpcijsko spektrofotometrijo iskali snovi, ki bi zmanjšale iztekanje laktoferina skozi ovojnice mikrosfer. Zadrževanje beljakovin je bilo najvišje ob dodatku 10 % laktoze in ob znižanju pH raztopine za premreženje. Z namenom ugotovitve optimalnih procesnih parametrov smo s pristopom načrtovanja eksperimentov izvedli 4-faktorski 2-nivojski popolni faktorski načrt. Eksperimenti so se razlikovali po deležu alginata, laktoferina in vrednosti pH raztopine CaCl2. Preučili smo, kako posamezen faktor vpliva na vsebnost laktoferina v mikrosferah. Alternativno pot razvoja formulacije z laktoferinom smo načrtovali s postopkom oblaganja pelet. V vrtinčnoslojni komori smo obložili peletna jedra iz mikrokristalne celuloze. Na nevtralna paletna jedra smo nanesli dve oblogi. Za dosego čim višje vsebnosti laktoferina smo sestavo posamezne obloge spreminjali. Želeli smo izdelati pelete, ki bodo v eni kapsuli vsebovale 200 mg laktoferina in bodo obložene z gastrorezistentno oblogo. Pelete smo vrednotili s farmakopejskimi metodami za testiranje kakovosti farmacevtskih oblik in preverjali ustreznost izdelanih formulacij. Najbolj optimalne pelete so nastale pri oblaganju s formulacijo 1. Preizkus sproščanja pelet smo opravili v sistemih za sproščanje farmacevtskih oblik II in III. Iz profilov sproščanja je razvidno, da se je iz pelet sprostilo le 80 % laktoferina. Možna razloga za nestabilnost laktoferina sta lokalno povišanje pH ob dodatku 0,2 M Na3PO4 ali hitra sprememba pH med nevtralizacijo medija. Uspešno smo izdelali formulacije z obema oblogama in pokazali, da je metoda oblaganja bolj perspektivna od mikrokapsuliranja.

• Language: Slovenian
• Keywords: laktoferin, načrtovanje eksperimentov, mikrokapsule, pelete, kapsule
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2021
• PID: 20.500.12556/RUL-132401
• Publication date in RUL: 25.10.2021

Secondary language

• Language: English
• Title: Development and evaluation of oral lactoferin formulations manufactured by microcapsulation and film-coating

Abstract

Lactoferrin is a glycoprotein which is excreted by numerous glands in the body and can also be found in milk. Its widespread presence in the body shows its a great significance of for the organysm. Its protecting benefits range from antimicrobial to antivirus and anti-cancerous activity. This wide range of activities is enabled by the mechanisms of lactoferrin action that show its capability of binding Ferrum with the cell components of the host and pathogens. Due to many positive benefits of lactofferin and with the purpose of lowering the environmental impact caused by the waste sour whey, we created a dietary supplement in the form of microspheres. For the creation of microspheres a method of dripping a solution of Natrium alginate with the assistance of peristaltic pump into a solution of Ca2+ ions was used. Microspheres of small sizes were made by an encapsulator Inotech IE-50R. Only a small amount of lactofferin is incorporated in alginate microspheres, therefore by using an absorption spectrophotometry we were searching for substances that would reduce lactoferrin leaking through the membranes of the microspheres. Retention of proteins was the highest when adding 10 % of lactose and reducing pH of a cross-linking solution. To find out the optimal process parameters, we started planning the experiments and performed a 4-factor 2-level complete factor plan. Experiments varied according to the amount of alginate, lactoferrin and the pH level of a cross-linking CaCl2 solution. We studied the impact of each factor on the content of lactoferrin in microspheres. The alternative way of developing formulation with lactoferrin was planned by the procedure of pellet coating. The pellets of microcrystalline cellulose were coated in a fluid bed pellet coater. Two coatings were applied on neutral pellet nuclei. In order to achieve the highest possible content of lactoferrin, the composition of each coating was changed. Our aim was to produce pellets with 200 mg of lactoferrin in one capsule and will have a gastro-resistant coating. For the evaluation of the pellets pharmacopeia methods for testing quality of dosage forms were used and the adequacy of the formulations was checked. The most optimal pellets were formed by coating with the formulation 1. The dissolution test of pellets was performed by the dissolution aparatuses II and III. The dissolution profile of releasing showed that only 80 % of lactoferrin had been released from the pellets. Possible reasons for lactoferrin instability are a local increase in pH with the addition of 0.2 M Na3PO4 or a rapid change in pH during neutralization of the medium. We have successfully developed formulations with both coatings, and showed that the coating method is more promising than microencapsulation.

• Keywords: lactoferrin, planning the experiments, microcapsules, pellets, capsules