EFFECT OF LAMOTRIGINE SOLID DISPERSIONS ON VIABILITY OF A549 AND RG-2 CELLS

Öz

Objective: LMT is an antiepileptic drug that is among the drugs being repositioned/reprofiled beyond its original medical indications, thanks to its promising anticancer activity profiles on various cancer cells. In this study, the improvement of the poor aqueous solubility of LMT using solid dispersion technique with different polymeric/surfactant carriers in different ratios and the antiproliferative effect potentials of the obtained optimum LMT solid dispersion on A549 and RG-2 cells were determined. Material and Method: LMT solid dispersions were prepared using four different carrier types (PEG 4000, PEG 6000, poloxamer 188 and poloxamer 407) and five different drug to carrier ratios (1:2 - 1:4 - 1:6 - 1:8 - 1:10) according to the melting method and the change in the solubility profile was determined by solubility studies performed according to the shake-flask method. The MTT assay protocol was used to determine the cytotoxicity profile of the selected optimum solid dispersion (F8) on A549 and RG-2 cell lines. The interaction of LMT with the formulation components and the solid-state characteristics within the formulation were evaluated by XRD, DSC, FTIR, and SEM analyses. Result and Discussion: The solubility studies showed that LMT exhibits a pH-dependent solubility profile, and the highest solubility increase (2.3 fold) in solid dispersion formulations was obtained with the F8 formulation using poloxamer 407 at a 1:6 ratio. XRD, DSC, FTIR, and SEM analysis results explain the partial amorphization of LMT in the F8 formulation. In vitro cytotoxicity studies have shown that LMT has a dose-dependent antiproliferative effect on RG-2 cells, but no significant cytotoxic effect at low doses (10-80 µg/ml). For A549 cells, cell viability decreased with increasing doses, suggesting promising results regarding antiproliferative effects at higher doses.

Anahtar Kelimeler

• Antiproliferative effect
• poloxamers
• polyethylene glycols
• lamotrigine
• solid dispersion
• solubility

Etik Beyan

The authors declare that there is no real, potential, or perceived conflict of interest for this article.

Teşekkür

SEM and XRD analyses were conducted at İnönü University Scientific and Technological Research Center (İBTAM), and DSC and FTIR analyses were performed at Erzincan Binali Yıldırım University Fundamental Sciences Application and Research Center. Author would like to thank Asst. Prof. Naile Öztürk and Abdulaziz Güneş for their kind help in experimental studies. Also, the author would gratefully like to thank Prof. Dr. Imran Vural and Prof. Dr. Selma Şahin for their support.

LAMOTRİJİN KATI DİSPERSİYONLARININ A549 VE RG-2 HÜCRELERİNİN CANLILIĞI ÜZERİNE ETKİSİ

Öz

Amaç: LMT, çeşitli kanser hücreleri üzerindeki ümit verici antikanser aktivite profilleri sayesinde orijinal tıbbi endikasyonlarının ötesinde yeniden konumlandırılan/yeniden profillenen ilaçlar arasında yer alan bir antiepileptik ilaçtır. Bu çalışmada LMT'nin zayıf sulu çözünürlüğünün farklı oranlarda farklı polimerik/yüzey aktif madde taşıyıcıları ile katı dispersiyon tekniği kullanılarak iyileştirilmesi ve elde edilen optimum LMT katı dispersiyonunun A549 ve RG-2 hücreleri üzerindeki antiproliferatif etki potansiyelleri belirlenmiştir. Gereç ve Yöntem: LMT katı dispersiyonları eritme yöntemine göre dört farklı taşıyıcı türü (PEG 4000, PEG 6000, poloksamer 188 and poloksamer 407) ve beş farklı ilaç:taşıyıcı oranı (1:2 - 1:4 - 1:6 - 1:8 - 1:10) kullanılarak hazırlanmış ve çözünürlük profilindeki değişim şişe çalkalama yöntemine göre yapılan çözünürlük çalışmaları ile belirlenmiştir. Seçilen optimum katı dispersiyonun (F8) A549 ve RG-2 hücre hatları üzerindeki sitotoksisite profilinin belirlenmesinde MTT deney protokolü kullanılmıştır. LMT'nin formülasyon bileşenleri ile etkileşimi ve formülasyon içindeki katı hal karakteristiği XRD, DSC, FTIR ve SEM analizleri ile değerlendirilmiştir. Sonuç ve Tartışma: Çözünürlük çalışmaları LMT'nin pH bağımlı bir çözünürlük profili sergilediğini göstermiş olup katı dispersiyon formülasyonlarında en yüksek çözünürlük artışı (2.3 kat) poloksamer 407'nin 1:6 oranında kullanıldığı F8 formülasyonu ile elde dilmiştir. XRD, DSC, FTIR ve SEM analiz sonuçları F8 formülasyonu içindeki LMT'nin kısmi amorfizasyonu açıklar niteliktedir. İn vitro sitotoksisite çalışmaları, RG-2 hücreleri için LMT'nin doza bağlı bir antiproliferatif etkisinin olmakla birlikte düşük dozlarda (10-80µg/ml) anlamlı bir sistotoksik etkiye sahip olmadığını göstermiştir. A549 hücreleri için, doz arttıkça hücre canlılığının azalması, daha yüksek dozlarda antiproliferatif etkiye ulaşılması ile ilgili umut verici sonuçlar alınabileceğini düşündürmektedir.

Anahtar Kelimeler

• Antiproliferatif etki
• çözünürlük
• katı dispersiyon
• lamotrigin
• polietilen glikoller
• poloksamerler

Etik Beyan

Yazarlar bu makale için gerçek, potansiyel veya algılanan bir çıkar çatışması olmadığını beyan ederler.

Teşekkür

SEM ve XRD analizleri İnönü Üniversitesi Bilimsel ve Teknolojik Araştırma Merkezi'nde (İBTAM), DSC ve FTIR analizleri ise Erzincan Binali Yıldırım Üniversitesi Temel Bilimler Uygulama ve Araştırma Merkezi'nde gerçekleştirilmiştir. Yazar, deneysel çalışmalardaki nazik yardımları için Dr. Öğretim Üyesi Naile Öztürk ve Abdulaziz Güneş'e teşekkür eder. Ayrıca, Prof. Dr. İmran Vural ve Prof. Dr. Selma Şahin'e de destekleri için minnettarlığını sunar.

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