<p>During oncogenesis, cancer cells go through metabolic reprogramming to maintain their high growth rates and adapt to changes in the microenvironment and the lack of essential nutrients. Several types of cancer are dependent on de novo fatty acid synthesis to sustain their growth rates by providing precursors to construct membranes and produce vital signaling lipids. Fatty acid synthase (FASN) catalyze the terminal step of de novo fatty acid synthesis and it is highly expressed in many types of cancers where it’s up-regulation is correlated with cancer aggressiveness and low therapeutic outcome. Many FASN inhibitors were developed and showed potent anticancer activity however, only one inhibitor advanced to early stage clinical trials with some dose limiting toxicities. Using a modified fluorescence-linked enzyme chemoproteomic strategy (FLECS) screen, we identified HS-106, a thiophenopyrimiden FASN inhibitor that has anti-neoplastic activity against breast cancer in vitro and in vivo. HS-106 was able to inhibit both; purified human FASN activity and cellular fatty acid synthesis activity as evaluated by radioactive tracers incorporation into lipids experiments. In proliferation and apoptosis assays, HS-106 was able to block proliferation and induce apoptosis in several breast cancer cell lines. Several rescue experiment and global lipidome analysis were performed to probe the mechanism by which HS-106 induces apoptosis. HS-106 was found to induce several changes in lipids metabolism: (i) inhibit fatty acids synthesis. (ii) Inhibit fatty acids oxidation as indicated by the ability of inhibiting Malonyl CoA accumulation to block HS-106 induced apoptosis and the increase in the abundance of ceramides. (iii) Increase fatty acids uptake and neutral lipids formation as confirmed 14C Palmitate uptake assay and neutral lipids staining. (iv)Inhibit the formation of phospholipids by inhibiting de novo fatty acid synthesis and diverting exogenous fatty acids to neutral lipids. All of these events would lead to disruption in membranes structure and function. HS-106 was also tested in Lapatinib resistant cell lines and it was able to induce apoptosis and synergizes Lapatinib activity in these cell lines. This may be due the disruption of lipid rafts based on the observation that HS-106 reduces the expression of both HER2 and HER3. HS-106 was found to be well tolerated and bioavailable in mice with high elimination rate. HS-106 efficacy was tested in MMTV neu mouse model. Although did not significantly reduced tumor size (alone), HS-106 was able to double the median survival of the mice and showed potent antitumor activity when combined with Carboplatin. Similar results were obtained when same combinations and dosing schedule was used in C3Tag mouse model except for the inability of HS-106 affect mice survival.</p><p>From the above, HS-106 represent a novel FASN inhibitor that has anticancer activity both in vivo and in vitro. Being a chemically tractable molecule, the synthetic route to HS-106 is readily adaptable for the preparation of analogs that are similar in structure, suggesting that, the pharmacological properties of HS-106 can be improved.</p> / Dissertation

Background: Chemotherapeutic agents directed against human epidermal growth factor receptor 2 (HER-2) have significantly improved the prognosis of patients who are positive for this receptor. However, cardiomyopathy remains as a common adverse effect of using these agents. Materials and Methods: Literature search was conducted via PubMed using the keywords of "Trastuzumab Cardiomyopathy," "Lapatinib Cardiomyopathy" and "Pertuzumab Cardiomyopathy," which provided 104 results. These articles were then screened for relevance to the targeted subject based on their title and abstracts. Case reports and articles that were not discussing any aspect of cardiomyopathy secondary to targeted therapy for breast cancer and articles not in English were eliminated. After elimination, a bibliography search among selected articles was done and a total of 46 articles were identified. The collected articles were then meticulously analyzed and summarized. Results: The use of human epidermal growth factor receptor 2 (HER-2) receptor targeted chemotherapy in breast cancer is limited because of a higher incidence (19-22%) of cardiomyopathy. The incidence of cardiomyopathy is not dose dependent and in most cases it is reversible after discontinuation of the drug and treatment with heart failure medications. Severe adverse outcomes including death or permanent disability are rare. Conclusion: HER-2 targeted chemotherapy for breast cancer has a higher incidence of associated reversible cardiomyopathy. Patients should be monitored by serial echocardiography starting at the beginning of the treatment and followed by every 3 months until the completion of chemotherapy. Co-ordination between oncologists and cardiologists is needed to develop evidence-based protocols to prevent, identify, monitor and treat trastuzumab-induced cardiomyopathy.

In the United States, breast cancer is the most commonly diagnosed cancer and is the second most common cause of cancer-related deaths among women. Among the various subtypes of breast cancer, 25-30% of diagnoses present themselves as human epidermal growth factor receptor 2 positive (HER-2+). HER-2 is a protein receptor located on the cell surface that interacts with other proteins and signaling molecules to translate extracellular signals into cellular process such as cell growth and replication. However, in breast cancer, there is a drastic increase in the number of HER-2 proteins on the cell surface, that causes excessive cell growth and proliferation, and ultimately tumor formation. The most frequent treatment of HER-2+ breast cancers includes the use of a single agent inhibitor that directly blocks the HER-2 protein to prevent over-signaling and cell growth. However, after continuous use, breast cancer cells develop drug resistance, as other proteins such as the insulin-like growth factor 1 receptor (IGF-1R) and the protein kinase B (AKT) can also interfere and cause cell growth and replication. In this study, we propose that the use of a multi-agent treatment targeting the HER-2, IGF-1R, and AKT proteins will be more effective than a single-agent treatment of HER-2 alone. Through protein analysis by mass spectrometry, we intend to illuminate the different cellular responses to both treatment types. The results indicate that the single drug treatment targeting Her-2 appears to increase processes related cellular repair, while the multi-drug treatment indicates an increase in processes related to programmed cell death; both treatments appear to block the transmission of protein signaling. / MS / In the United States, breast cancer is the most commonly diagnosed cancer and is the second most common cause of cancer-related deaths among women. Among the various subtypes of breast cancer, 25-30% of diagnoses present themselves as human epidermal growth factor receptor 2 positive (HER-2+). HER-2 is a protein receptor located on the cell surface that interacts with other proteins and signaling molecules to translate extracellular signals into cellular process such as cell growth and replication. However, in breast cancer, there is a drastic increase in the number of HER-2 proteins on the cell surface, that causes excessive cell growth and proliferation, and ultimately tumor formation. The most frequent treatment of HER-2+ breast cancers includes the use of a single agent inhibitor that directly blocks the HER-2 protein to prevent over-signaling and cell growth. However, after continuous use, breast cancer cells develop drug resistance, as other proteins such as the insulin-like growth factor 1 receptor (IGF-1R) and the protein kinase B (AKT) can also interfere and cause cell growth and replication. In this study, we propose that the use of a multi-agent treatment targeting the HER-2, IGF-1R, and AKT proteins will be more effective than a single-agent treatment of HER-2 alone. Through protein analysis by mass spectrometry, we intend to illuminate the different cellular responses to both treatment types. The results indicate that the single drug treatment targeting Her-2 appears to increase processes related cellular repair, while the multi-drug treatment indicates an increase in processes related to programmed cell death; both treatments appear to block the transmission of protein signaling.

La P-glycoprotéine (P-gp) est une protéine transmembranaire de la famille des ATP binding cassette transporteurs. Elle est impliquée dans l’efflux du milieu intracellulaire vers le milieu extracellulaire d’une grande variété de médicaments anticancéreux. Elle peut être responsable de la diminution de la biodisponibilité orale et de la concentration intra-tumorale des médicaments qui en sont substrats. Elle peut notamment être surexprimée par les cellules cancéreuses des adénocarcinomes du colon naïfs de tout traitement, suggérant une résistance naturelle de cette tumeur et également après une chimiothérapie. Notre premier travail in vivo a documenté le caractère substrat de la P-gp de l’evérolimus, inhibiteur de mTOR indiqué dans divers cancers (rein, tumeurs neuroendocrines d’oringine pancréatique et sein), jusqu’à maintenant uniquement étudié dans des modèles in vitro. Une augmentation significative de l’AUC de l’evérolimus administré par voie orale est observée chez des souris mdr1a-/b- comparées à des souris mdr1a+/1b+. Une amélioration significative de la biodisponibilité orale de l’evérolimus est aussi notée chez des souris prétraitées par le lapatinib (Tyverb®), inhibiteur des tyrosines kinases (EGFR et HER2) indiqué dans le cancer du sein, par rapport aux souris ayant reçu l’evérolimus seul. Ce résultat est accompagné d’une inhibition de l’expression de la P-gp intestinale par le lapatinib mesurée par la technique de Western Blot. Enfin, une étude préclinique menée chez des souris porteuses d’une xénogreffe colorectale mutée KRAS montre une activité anti-tumorale certaine des deux médicaments utilisés seuls et en schéma séquentiel. Notre seconde étude a montré pour la première fois que le cétuximab (Erbitux®), anticorps anti-EGFR, inhibe la fonctionnalité de la P-gp dans deux lignées cellulaires surexprimant la P-gp (les cellules IGROV-1 et les HEK P-gp) indépendamment de leur statut EGFR et entraîne chez des souris porteuses d’une xénogreffe colorectale une augmentation significative de la biodisponibilité orale et de la concentration intra-tumorale du SN-38, métabolite actif de l’irinotécan (Campto®) administré par voie orale. Le cétuximab étant prescrit en association avec l’irinotécan chez des patients atteints d’un cancer colorectal métastasé, initialement réfractaire à l’irinotécan, ces résultats pourraient en partie expliquer la réversion de la résistance à l’irinotécan par le cétuximab par une inhibition de l’efflux de la P-gp. Grâce à l’étude de deux associations de médicaments «lapatinib-evérolimus» et «cétuximab-irinotécan», nous avons démontré l’intérêt de l’étude de l’inhibition de la P-gp avec les traitements les plus récents, notamment son rôle dans l’amélioration de la biodisponibilité orale de chimiothérapies utilisées par voie orale. / P-glycoprotein (P-gp) is a membrane transporter and belongs to the ATP-binding cassette (ABC) transporter super family. P-gp decreases oral bioavailability of substrate drugs and can cause multidrug resistance in tumor cells by decreasing intracellular drug levels. P-gp is overexpressed in colorectal carcinoma naturally resistant to chemotherapy. The aim of our first study was to document the in vivo transport of everolimus (Afinitor®), a mTOR inhibitor, by P-gp. A significant increase of everolimus oral bioavaibility was observed in mdr1a-/1b- mice compared to the wild type. In addition, a significant increase of everolimus oral bioavaibility was showed in mice that received a lapatinib pre-treatment (a dual EGFR/HER2 tyrosine kinase inhibitor) compared to mice that received everolimus alone. These results were accompanied by a significant decrease of P-gp expression in duodenum segment in lapatinib pre-treated group as compared to control group. Finally, each drug given alone or in association showed a major antitumor activity in a xenograft model of human colorectal carcinoma with KRAS mutation. Our second study showed for the first time that cetuximab (Erbitux®), a monoclonal antibody directed towards EGFR, inhibits P-gp functionality in two cell lines overexpressing P-gp (IGROV-1 and HEK P-gp cells) independently of EGFR status and leads to significant increases of oral bioavailability and intratumoral concentration of SN-38, the active metabolite of irinotecan (Campto®) in mice bearing colorectal carcinoma xenograft. Cetuximab is used in combination with irinotecan in patients with metastatic colorectal cancer, initially refractory to irinotecan, our results may partly explain the reversion of resistance to irinotecan by inhibiting P-gp efflux by cetuximab. In conclusion, our results showed the interest to study the effect of recent anticancerous drugs on P-gp, including their ability to improve oral bioavailability of oral chemotherapy used.

Les résultats des projets actuels apportent une information, sur différents aspects des rôles de la voie PI3K, dans le développement du cancer du sein, et la réponse au traitement. Les projets particuliers couvrent des sujets liés à la voie aux niveaux concernant les récepteurs de la famille HER, activant la voie PI3K, ainsi que PI3K et les effecteurs en découlant. Les effets pronostic et prédictif de la dérégulation de PI3K sont les sujets centraux de la recherche décrite ici. Une baisse d’expression de PI3KR1 est associée à une survie réduite dans notre cohorte de patients. Une attention particulière a été portée aux mutations de PIK3CA communes dans le cancer du sein. Tandis que les mutations de PIK3CA agissent comme des marqueurs de bon pronostic chez les patients anti-HER2-naïfs, ces mutations agissent au contraire comme prédicteurs négatifs de la réponse au traitement par trastuzumab. Les résultats décrits mènent un peu plus vers l’implication de plusieurs voies moléculaires altérées, en particulier la voie de signalisation Wnt, dans la tumorigénèse des cancers du sein PIK3CA mutés. De plus, nous avons testé les taux de lapatinib plasmatique montrant une augmentation pertinente dans les périodes d’état d’équilibre du traitement. Par ailleurs, nous avons démontré des incohérences dans l’évaluation de l’EGFR et proposé des approches pour l’interprétation des comptages d’immunohistochimie et de FISH. Tous ces sujets sont connectés par la 170 voie PI3K, et le besoin d’approfondir les connaissances actuelles, et d’apporter de nouvelles informations utiles applicables dans le futur dans les pratiques cliniques / Results of the presented research projects bring information about several aspects of the PI3K signaling pathway roles in breast cancer development and treatment response. The particular projects covered the subjects connected with the signaling pathway, ranging from the HER family receptors activating the pathway, and PI3K to the downstream levels of signalisation. The prognostic and predictive effect of PI3K deregulation was the central subject of the described research. The decreased expression of PIK3R1 associated with reduced survival of our patients. A special focus was put on the PIK3CA mutations which are common in breast cancer. Whereas the PIK3CA mutations act as a good prognostic marker in patients non-treated with the HER2 inhibitors, these mutations predict a negative response to trastuzumab treatment. The described results, furthermore, draw attention to the role of several altered molecular signaling pathways in breast cancer development, especially to the Wnt signaling pathway. The lapatinib plasma levels showing the relevant increase in comparison with the already described efficient steady-state levels were also described in one of the projects. Moreover, various modifications to EGFR status assessment were compared and showed that EGFR FISH and IHC count interpretation depended significantly on method and thresholds used. All these subjects are connected by the PI3K pathway, the need to deepen current knowledge and bring new useful information applicable in future clinical practice.

Les thérapies ciblées telles que les inhibiteurs de la tyrosine kinase ont permis d'améliorer le traitement du cancer du sein en ciblant HER2. Cependant, il a été observé que le lapatinib était moins efficace chez les patients obèses ou en surpoids que chez les patients de poids normal.Nous avons d'abord reproduit l'effet de résistance des cellules de cancer du sein au lapatinib en présence de tissu adipeux tel qu’il a été observé pour d’autres thérapies. Les cellules tumorales qui surexpriment HER2 étaient partiellement résistantes au lapatinib mais également à d'autres inhibiteurs de la tyrosine kinase lorsqu'elles étaient en contact avec le milieu conditionné d’adipocytes. En implantant du tissu adipeux humain et des tumeurs humaines chez la souris, nous avons pu étudier la résistance du cancer du sein au lapatinib in vivo.Pour comprendre le mécanisme de cette résistance, nous avons exposé les adipocytes à plusieurs modulateurs du métabolisme. La cytotoxicité cellulaire induite par le lapatinib était plus faible pour les cellules tumorales exposées à un milieu conditionné d'adipocytes préalablement incubés avec les alpha-bloquants qu'à un milieu conditionné à partir d'adipocytes seuls. De la même manière, cette toxicité était inférieure pour les agonistes des récepteurs alpha adrénergiques, pour les bêtabloquants et pour les inhibiteurs de lipolyse. Au contraire, la cytotoxicité a été augmentée pour les cellules tumorales en contact avec le milieu conditionné d’adipocytes exposés aux agonistes du récepteur bêta-adrénergiques. Au niveau des cellules cancéreuses, l'arrêt du cycle cellulaire induit par le lapatinib était réduit pour les cellules tumorales exposées au milieu conditionné d’adipocytes en ce qui concerne le pourcentage de cellules dans la phase G0/G1. Ceci s’est vérifié en étudiant l'expression des gènes codant pour plusieurs protéines impliquées dans la progression du cycle cellulaire / Targeted therapies as tyrosine kinase inhibitors permitted an improvement of breast cancer therapies by targeting HER2. However, resistance has been observed in obèse patients for lapatinib treatment.We reproduced the effect of resistance of breast cancer cells to laaptinib in presence of adipose tissue as observed for other therapies. Tumor cells overexpressing HER2 was partly resistant to lapatinib but also for other tyrosine kinase inhibitors when in contact with adipocyte-conditioned medium. By impnating human adipose tissue nad human tumors in mice, we were able to study rhe resistance of breast tumor cells in vivo.In order to elucidate the mechanism of such resistance, we exposed the adipocytes to several metabolism modulators. The lapatinib-induced cell cytotoxicity was lower for the tumor cells exposed to the conditioned medium from adipocytes earlier exposed to alpha blockers than to the conditioned medium from adipocytes alone. In the same manner, the toxicity was lower for the agonists of alpha-adrenergic receptors , for beta-blockers and for the lipolysis inhibitors. At the opposite, the cytotoxicity was enhanced for tumor cells in contact with the conditioned medium of adipocytes exposed to the agonists of beta adrenergic receptors.At the tumor cell level, the laaptinib-induced cell cycle arrest was reduced for the tumor cells exposed to the conditioned medium regarding the G0/G1 phase. That was verified by the study of the expression of genes involved in the cell cycle progression

Glioblastoma multiforme (GBM) is the most common and malignant brain tumor in adults, affecting thousands of people worldwide every year, with a life expectancy, post diagnosis of 12 months. Surgery, radiotherapy and chemotherapy together, result in an overall mean survival not exceeding 15 months. Targeted therapeutic agents sorafenib, an oral multi kinase inhibitor, and lapatinib, an epidermal growth factor receptor (EGFR) inhibitor, used in combination have been shown to kill GBM cells be through inhibition of major growth mediating signaling pathways that are frequently over expressed in gliomas, including mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase/ protein kinase B (PI3K/AKT). Sorafenib can restore lapatinib induced cytotoxicity by down regulation of myeloid cell leukaemia-1 (Mcl-1) expression. Prior studies have shown Mcl-1 to play an important role in resistance to lapatinib. Furthermore, data indicated that this drug combination is able to trigger activation of autophagic and apoptotic pathways and induce endoplasmic reticulum (ER) stress response in GBM cells, collectively resulting in cell death. In conclusion, data presented here demonstrates that the combination of sorafenib and lapatinib can kill GBM cells in a greater than additive fashion, through induction of autophagy, apoptotic events (extrinsic and intrinsic) and ER stress.

We investigated the mechanism of toxicity and resistance development of small molecule tyrosine kinase inhibitor lapatinib in HCT 116 colon cancer cells. Lapatinib mediated cell death in HCT 116 cells was caspase independent and involved cytosolic release of apoptosis inducing factor. Treatment of HCT 116 cells with 10µM Lapatinib lead to the outgrowth of lapatinib resistant HCT 116 cells. Our studies show that alterations in the expression and activation of Bcl-2 family proteins allow lapatinib resistant HCT 116 cells to resist cytotoxic effects of lapatinib as well as of other commonly used chemotherapeutic agents. In hepatoma and pancreatic cancer cells, the effects of combining multi-kinase inhibitor sorafenib with histone deacetylase inhibitors (HDACIs) namely, vorinostat and sodium valproate were investigated. It was found that sorafenib synergizes with HDACIs resulting in enhanced cell death compared to death induced by the drugs individually. The mechanism of action of sorafenib and vorinostat combination treatment as well as sorafenib and sodium valproate combined treatment was shown to involve activation of the CD95 death receptor pathway. Alterations in the CD95 pathway can render cancer cells resistant to chemotherapeutic agents. Hence, we combined sorafenib+sodium valproate with a BH-3 domain mimetic named obatoclax (GX-15-070) which resulted in enhanced toxicity to cancer cells. More importantly, knock-down of CD95 (to mimic non-functional CD95 pathway) reduced cell death induced by sorafenib+sodium valproate combined but failed to protect cells from cell death induced by sorafenib+sodium valproate+obatoclax combined. This suggests that combining sorafenib+HDACI with obatoclax may not only enhance toxicity to cancer cells but may also reduce chances of resistance development via alterations in the CD95 pathway. These studies enhance our knowledge of existing treatment strategies for cancer as well as throw light on how current approaches can be improved in order to better diagnose and treat cancer. Understanding mechanisms of drug action as well as resistance development will allow us to combine existing therapies effectively in order best target cancer cells as well as provide us with information that can help us design new cancer treatment strategies.

The cancers of liver, colon and breast are amongst the top five most prevalent and most fatal worldwide. As the Raf/MEK/ERK pathway is frequently deregulated in hepatocellular carcinoma (HCC), sorafenib, a Raf kinase inhibitor, became the first systemic therapy approved for the treatment of patients with HCC. However, sorafenib only produced modest effects with low response rates in the clinic. Similarly, regorafenib, which was approved for the treatment of metastatic colorectal cancer (CRC), has had a poor response rate in the clinic. Since phosphodiesterase type 5 has been reported to be overexpressed in HCC and CRC, we hypothesized that sildenafil, a phosphodiesterase type 5 inhibitor, could enhance the toxicities of sorafenib and regorafenib in HCC and CRC cells, respectively. Our in vitro data indicated that the drugs interacted strongly to kill cancer cells via induction of ER stress, autophagy and apoptosis. In accordance with these findings, our in vivo data demonstrated a significant reduction in tumor growth. The second study in this manuscript was conducted based on the growing body of evidence about the significant contribution of EGFR and JAK/STAT signaling to the breast tumorigenesis. Our preliminary in vitro data demonstrated that the concurrent inhibition of these two pathways by lapatinib, a dual ERBB1/2 inhibitor, and ruxolitinib, a JAK1/2 inhibitor, synergistically killed breast cancer cells of all types, including the resistant triple negative subtype. Our mechanistic studies showed that the combination of ruxolitinib and lapatinib triggered cytotoxic mitophagy, and autophagy-dependent activation of BAX and BAK leading to the mitochondrial dysfunction.

Tumores de mama podem ser categorizados pela expressão de diferentes receptores, como por exemplo o HER-2 (receptor de fator de crescimento epidérmico 2), sendo amplamente estudada a utilização do fármaco inibidor do receptor da tirosina–quinase, ditosilato de lapatinib. Nesse sentido, o trabalho está centrado no desenvolvimento farmacotécnico e caracterização físico-química de nanocápsulas multiparede revestidas com polissorbato 80 e fosfatidilcolina (LIPOID S75®), seguidas de revestimento com quitosana, contendo ditosilato de lapatinib (25μg/mL e 50μg/mL). Estudos físico-químicos da fase orgânica e das nanocápsulas contendo LIPOID S75®, em diferentes concentrações, foram desenvolvidos através de medidas de viscosidade, tensão superficial e analise de calorimetria exploratória diferencial, correlacionando com o tamanho de partícula e interação da fosfatidilcolina no sistema. A partir disso, escolheu-se a maior concentração de LIPOID S75® testada, para desenvolver as nanocápsulas multiparede complexadas com metal e fenilalanina contendo ditosilato de lapatinib (Lap25-P5-Q-Fe-Phe; Lap50-P5-Q-Fe-Phe; Lap25-P5-Q-Zn-Phe; Lap50-P5-Q-Zn-Phe). As formulações contendo fármaco apresentaram distribuição unimodal de partículas na faixa nanométrica, (111 - 172 nm) e baixo índice de polidispersão. A faixa de potencial zeta obtida (+16 - +22 mV) indicou o revestimento da partícula pela quitosana. O ditosilato de lapatinib foi quantificado por cromatografia líquida de alta eficiência (HPLC). A fase móvel do método consistiu em acetonitrila:acetato de amônio 0,02M (70:30), vazão de 1,0 mL/min com detecção em 260 nm. As suspensões contendo 25 μg/mL e 50 μg/mL de fármaco apresentaram teor entre 93% e 91%, respectivamente. A eficiência de encapsulação (EE%) foi acima de 90%. O conjunto destes estudos demonstra que nanocápsulas desenvolvidas são potenciais candidatas para ensaios biológicos in vitro. / Breast tumors may be categorized by the expression of different receptors such as HER-2 (growth factor epidermal receptor 2), The use of the inhibitor drug receptor tyrosine-kinase lapatinib ditosylate (Tyverb®, GlaxoSmithKline) has been widely studied. In this sense, the work is focused on pharmaceutics development and physicochemical characterization of multiwall nanocapsules coated with polysorbate 80 and phosphatidylcholine (Lipoid S75®), followed by coating with chitosan containing lapatinib ditosylate (25 μ/mL and 50 μg/mL). Physicochemical studies of the organic phase and nanocapsules containing Lipoid S75® at different concentrations were developed by viscosity, surface tension analysis and differential scanning calorimetry, correlated with the particle size and interaction of phosphatidylcholine in the system From this, we have chosen the highest concentration of lipoid S75® tested to develop complexed nanocapsules multiparede with metal and phenylalanine containing lapatinib ditosylate (Lap25-P5-Phe-Phe-Q; Lap50-P5-Q -Fc-Phe; Lap25-P5-O-Zn-Phe; Lap50-P5-Zn-O-Phe). Formulations containing lapatinib ditosylate showed unimodal distribution of particles in the nanometer range (111-172 nm) and low polydispersity index. The obtained zeta potential range (+16 - +22 mV) indicated by the chitosan particle coating. The lapatinib ditosylate was quantified by high-performance liquid chromatography (HPLC). The mobile phase consisted method in acetonitrile: 0.02M ammonium acetate (70:30), flow 1.0 mLmin-1 with detection at 260 nm. The suspensions containing 25 ugmL-1 and 50 ugmL-1 showed drug content of between 93% and 91%, respectively. The encapsulation efficiency (EE%) was above 90%. Together these studies demonstrate that the nanocapsules are potential candidates for in vitro biological assays.