Diss. (sammanfattning) Uppsala : Univ., 2003. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Univ., 2003. / Härtill 4 uppsatser.

Cilengitide was anticipated to be a highly promising potential anti-angiogenic small molecule drug but unfortunately it failed phase III clinical trial in early 2013 (survival rate not passed). Today, there is still not a small molecule αvβ3 integrin drug in use in the clinic, new antagonists were urgently needed to advance the treatment of the αvβ3 integrin related diseases. / Peptide-type antagonists possess unique advantages such as non-accumulative, bio-friendly etc. comparing to other types of antagonists (peptidomimetics, non-peptidic etc.). Cilengitide is one of the peptide-type antagonists and it remains the most successful drug candidate demonstrated. Reinvestigation of the binding between Cilengitide and the αvβ3 integrin is therefore still meaningful. Specifically, the present study will concentrate on revealing the factors responsible for higher binding affinity of Cilengitide, comparing to the parent compound c(RGDfV), c(RADfV) and other N-methylated derivatives of c(RGDfV). Note that c(RADfV) could be also viewed as a methylation product of c(RGDfV) (methylation of side chain of the -G- residue). It is intended that the results of this investigation could provide clear guidance towards future efforts in the design of new peptide αvβ3 integrin antagonists. / Utilizing a novel but successful computational protocol, namely an integrated docking, molecular dynamics simulation and MM_GBSA free energy calculation method, the present thesis found that the binding profile between Cilengitide and the αvβ3 integrin is unique when compared to bindings of other methylation compounds. On the one hand, structural analysis (such as RMSD, ligand structure, hydrogen bond, backbone and side chain orientation) revealed that only c(RGDfV) and Cilengitide substantially retain the binding mode of the parent compound c(RGDfV) (Cilengitide is closer than c(RGDfV)). On the other hand, calculated binding energy results revealed that only c(RGDfV) and Cilengitide bind more strongly than c(RGDfV) to the αvβ3 integrin. Since in experiments only Cilengitide binds more strongly than c(RGDfV), it is therefore believed that both structural and energetic factors are responsible for the high binding affinity of Cilengitide. / Other energetic study revealed that the high binding affinity of Cilengtide compared to c(RGDfV) originates from Coulombic interaction (sum of electrostatic interaction and polar solvation energy), while van der Waals interaction and non-polar solvation energy was not favorable for Cilengitide binding. For the residue contribution, energy changes on the -R- and -G- residues upon methylation were nearly zero. Changes on the -D- and -f- residues were favorable for Cilengitide binding, while change on the -V- residue was not. Total changes on the five residues are favorable however. Pair-wise analysis suggests that interactions of the -D- residue in Cilengitide were very important for the binding, as suggested by its sizeable coupling energies with other residues. Amongst them the coupling between -D- and Mg688 is the most important pair. / In brief, the present study provides a quantitative understanding towards the binding between Cilengitide series ligand and the αvβ3 integrin. Through comparison with bindings between the methylated analogues of c(RGDfV), important features responsible for high binding affinity of Cilengitide were revealed. Some of the results contained in the thesis are in the process of being reported in "C. Yan, S. C. F. Au-Yeung. Investigation of the 'Methyl Effect' in the Cilengitide Binding Ligand Series and Its Implications on Future Integrin Antagonist Design. J. Med. Chem., in preparation (2014)". / Cilengitide是一种曾被高度寄望成为抗血管增生小分子药物的化合物。遗憾的是，2013年初所述化合物没有通过临床三期测试 (存活率不过关)。由于至今临床上仍然没有αvβ3整合素的小分子药物在使用，新的拮抗剂需要被设计出来以推进αvβ3整合素相关疾病的治疗。 / 相比于其它类型的拮抗剂 (类肽物、非肽类等)，多肽型的拮抗剂具有其独特的优点，譬如不积聚、生物友好等。Cilengitide是一种多肽型的拮抗剂，至今为止其仍是已证明的最成功的药物候选。重新研究Cilengitide与αvβ3整合素的结合情况因此仍然具有意义。具体地，本研究将聚焦于揭示相比于母体化合物c(RGDfV)、c(RADfV) 和c(RGDfV) 其它N-甲基化衍生物Cilengitide高亲和性的因素。c(RADfV) 可视为c(RGDfV) 另一甲基化的产物（-G- 残基侧链甲基化）。本研究的结果希望能给未来新型多肽型αvβ3整合素拮抗剂的设计以清晰的指导。 / 通过采用一种新的但颇成功的计算协议，即一种分子对接、分子动力学和MM_GBSA的综合方法，本论文发现相比于上述非Cilengitide的配体，Cilengitide与 αvβ3整合素的结合模式非常独特。一方面，结构分析 (RMSD、配体结构、氢键和骨架及侧链取向) 显示只有c(RGDfV) 与Cilengitide保留了其母体化合物，即c(RGDfV) 的结合模式（Cilengitide比c(RGDfV) 更接近）。另一方面，结合能计算结果显示只有Cilengitide和c(RGDfV) 与αvβ3整合素的结合比母体化合物c(RGDfV) 更强。由于实验上只有Cilengitide的结合强于c(RGDfV)，因此有理由相信结构和能量因素均对Cilengitide高亲和性负责。 / 其它能量研究显示相比于c(RGDfV)，Cilengitide的高亲和性来源于库仑作用 (静电作用和极性溶剂化能的加和)。范德华作用和非极性溶剂化能则对Cilengitide结合不利。残基贡献方面，甲基化后 -R- 和 -G- 的能量变化基本为零；-D- 和 -f- 残基的能量变化则有利于Cilengitide的结合，而 -V- 能量变化则不利。然而这五个残基的总变化对结合是有利的。配对分析结果显示Cilengitide中 -D- 残基的作用对于结合是非常重要的，这从其与其它残基可观的耦合能可以得知。其中，-D- 与Mg688的耦合是最重要的一对。 / 简言之，本研究提供了对Cilengitide系列配体与αvβ3整合素结合的定量理解。通过c(RGDfV) 甲基化后同类物的结合相互间的比较，揭示了负责Cilengitide高亲和性的重要特征。本论文中的一些研究结果正将报道于： / “严长青，欧阳植勋。关于Cilengitide配体系列中“甲基效应”的研究及其对未来αvβ3整合素拮抗剂设计的启示。药物化学杂志，准备中(2014)” / Yan, Changqing. / Thesis Ph.D. Chinese University of Hong Kong 2014. / Includes bibliographical references (leaves 138-146). / Abstracts also in Chinese. / Title from PDF title page (viewed on 18, January, 2017). / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only.

Thesis (Ph.D.)--University of Alabama at Birmingham, 2009. / Title from first page of PDF file (viewed on June 10, 2009). Includes bibliographical references.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A talidomida, anteriormente usada para aliviar desconfortos gÃstricos da gravidez, atualmente à usada para tratamento do cÃncer e de doenÃas inflamatÃrias. VÃrios anÃlogos ftalimÃdicos da talidomida tem sido pesquisados quanto a sua atividade antiangiogÃnica e imunomodulatÃria. Em estudos prÃvios, estes anÃlogos mostraram ausÃncia de atividade imunossupressora. O trabalho determinou, inicialmente, a atividade citotÃxica, de oito anÃlogos ftalimÃdicos da talidomida, frente a linhagens tumorais e de cÃlulas mononucleadas isoladas de sangue perifÃrico (CMSP) apÃs 72h de incubaÃÃo. Nenhum dos compostos desencadeou atividade citotÃxica in vitro, induÃÃo de hemÃlise em eritrÃcitos e potencial em induzir dano à fita de DNA, portanto nÃo foram genotÃxicos em CMSP humanas. O efeito antitumoral (in vivo) da talidomida e dos oito anÃlogos foi analisado em camundongos transplantados com o tumor Sarcoma 180 e tratados na dose 50mg/Kg/7 dias, via i.p. A inibiÃÃo do crescimento tumoral foi significante apenas para talidomida (53,5%) e anÃlogos SC-10 e SC-11 (67,9% e 67,4%, respectivamente), p<0,05. A anÃlise histopatolÃgica dos ÃrgÃos dos animais mostrou que a talidomida e seus anÃlogos provocam efeitos tÃxicos moderados, principalmente no fÃgado, mas esses podem ser considerados como reversÃveis. Os estudos acerca do mecanismo de aÃÃo foram aprofundados usando cÃlulas isoladas do Sarcoma 180, apÃs 72 horas de incubaÃÃo, nas doses de 10, 50 e 100Âg/mL, por citometria de fluxo, atravÃs dos seguintes ensaios: 1- AvaliaÃÃo da integridade de membrana, viabilidade celular e concentraÃÃo de cÃlulas; 2- DeterminaÃÃo do conteÃdo de DNA nuclear da cÃlula. Todos os compostos induziram a perda de integridade de membrana celular e fragmentaÃÃo internucleossomal do DNA nas maiores concentraÃÃes, indicando presenÃa de cÃlulas apoptÃticas em estÃgios finais ou em processo de necrose secundÃria. A avaliaÃÃo do potencial antiangiogÃnico, pelo ensaio do Wound Healing revelou que para a linhagem endotelial (HUVEC) o anÃlogo SC-10 causou uma inibiÃÃo maior (65,28%  1,58), enquanto que na linhagem tumoral o efeito maior foi do anÃlogo SC-11 (98,51%  0,25). A talidomida nÃo foi capaz de reduzir a migraÃÃo celular em nenhuma das linhagens testadas. No ensaio da membrana corioalantÃide (CAM), potencial antiangiogÃnico foi observado para os anÃlogos SC-10 e SC-11(5mg/mL), que inibiram o nÃmero de vasos (12, 88%  2,3 e 14,81%  3,3), a Ãrea de neovascularizaÃÃo (13,14%  1,7 e 14,26%  1,7) e o comprimento total de vasos em mm2 (9,19%  1,5 e 9,86%  1,9). A talidomida, nÃo foi capaz de inibir a vascularizaÃÃo embrionÃria. O efeito antitumoral (in vivo) da talidomida e anÃlogos SC-10 e SC-11 foi analisado em camundongos transplantados com o tumor Sarcoma 180, tratados na dose 50mg/Kg/10 dias, via i.p. A inibiÃÃo do crescimento tumoral para a talidomida (56.6%) e para os anÃlogos SC-10 e SC-11 (48,2% e 41,3%, respectivamente), p<0,05. A imunocoloraÃÃo de cÃlulas endoteliais intratumorais com CD-31, revelou-se, na forma de densidade microvascular, diminuÃda nos grupos tratados com anÃlogos SC-10 e SC-11 (64,59% e 46,51%), mas nÃo nos grupos tratados com a talidomida. Estes resultados, unidos a estudos prÃvios de imunomudulaÃÃo sugerem imunidade antitumoral e antiangiogÃnica, dependente de cÃlulas T, para os anÃlogos ftalimÃdicos. / The thalidomide was previously used to relieve gastric discomforts during pregnancy, and currently it is used for cancer treatment and inflammatory illnesses. Various phthalimides analogues of thalidomide have been researched for its antiangiogenic and immunemodulatory activity. In previous studies, these analogues showed absence of immunesuppressor activity. In this context, this work initially determined the cytotoxic activity of eight phthalimides analogues of thalidomide, in a series of cancer cell lines and of isolated peripheral mononuclear blood cells (PMBC) after 72h of incubation. None of compounds revealed cytotoxic activity in vitro in cancer cell lines and hemolytic activity towards of PBMC. They also showed no potential damage to the DNA strand, therefore they were not considered genotoxic towards human PMBC. The antitumor effect (in vivo) of thalidomide along with eight analogues was analyzed in mice transplanted with the Sarcoma 180 tumor and treated in a dose of 50mg/Kg/7 days, i.p. The inhibition of the tumor growth was only significant in mice treated with thalidomide (53.5%) and the analogues SC-10 and SC-11 (67.9% and 67.4%, respectively), p< 0,05. The histopathological analysis of the animalsâ organs showed mainly that thalidomide and its analogues cause moderate toxic effects, mostly in the liver, but these can be considered reversible. The studies concerning the mechanism of action were deepened using isolated cells from the Sarcoma 180, after 72 hours of incubation, in the doses of 10, 50 and 100Âg/mL. The following flow cytometry assays were carried out: 1- Evaluation of the membrane integrity, cellular viability and concentration of cells; 2- Determination of the nuclear DNA content. All the compounds led to the loss of membrane cell integrity and it was found internucleossomal DNA fragmentation in the higher concentrations, indicating the presence of cells with late stage apoptotic characteristics or in the process of secondary necrosis. The evaluation of the antiangiogenic potential, with the Wound Healing assay revealed that for the endothelial cell line (HUVEC) the analogue SC-10 caused a greater inhibition (65.28%  1,58), whereas in the tumor cell line the highest effect was of the analogue SC-11 (98.51%  0,25). The thalidomide was not capable of reducing cellular migration in none of the tested cell lines. In the chorioallantoic membrane assay (CAM), an antiangiogenic potential was observed with analogous SC-10 and SC-11 (5mg/mL), where there was an inhibition in the number of vessels (12, 88%  2,3 and 14.81%  3,3), the area of neovascularization (13.14%  1,7 and 14.26%  1,7) and the total length of vessels in mm2 (9.19%  1,5 and 9.86%  1,9). The thalidomide was not capable of inhibiting embryonic vascularization. The antitumor effect (in alive) of thalidomide and the analogues SC-10 and SC-11 was analyzed in mice transplanted with the Sarcoma 180 tumor, treated with the dose of 50mg/Kg/10 days, i.p. It was observed inhibition of the tumor growth in the thalidomide treatment (56,6%) and for analogues SC-10 and SC-11 (48.2% and 41.3%, respectively), p< 0,05. The immunostaining of intratumor endothelial cells with CD-31, showed, in the form of microvascular density, reduction in the groups treated with analogues SC-10 and SC-11 (64.59% and 46.51%), but not in the groups treated with thalidomide. These results, along with previous studies of immunemodulation suggest antitumor and antiangiogenic immunity, T cells dependent, for the phthalimides analogues.

Le cancer est l’une des principales causes de mortalité en France, après les maladies cardiovasculaires. Il est responsable de plus de 11 millions de décès dans le monde chaque année. Le cancer résulte d’une prolifération anarchique de cellules qui mène à la formation d’une tumeur. Les cellules tumorales peuvent ensuite migrer vers d’autres tissus pour former des métastases. La chimiothérapie est l’un des traitements les plus utilisés pour traiter le cancer. Elle consiste en l’utilisation d’agents antitumoraux qui provoquent la mort cellulaire en bloquant la mitose. Dans le but d’induire cette apoptose, nous nous sommes intéressés aux poisons du fuseau mitotique, agents cytotoxiques qui ont pour cible les microtubules et qui ont la particularité de se fixer sur leur constituant majeur, la tubuline. La dynamique des microtubules joue un rôle crucial dans la multiplication cellulaire. Bloquer cette dynamique est suffisant pour bloquer la mitose. Par ailleurs, suite à cet arrêt de la polymérisation, un second mécanisme se mettrait en place, notamment au niveau des cellules endothéliales, pour empêcher la néovascularisation, ce qui inhiberait ainsi l’angiogénèse. Notre travail consiste en la conception et la synthèse de nouveaux inhibiteurs de la polymérisation de la tubuline, potentiellement anti-angiogéniques et anti-vasculaires. Il s’agit de tricycles, qui ont la particularité d’interagir spécifiquement avec le site de fixation de la colchicine, au niveau de la tubuline, ce qui inhibe la polymérisation des microtubules et par conséquent la division cellulaire. Des tests d’inhibition enzymatique et de cytotoxicité sur plusieurs lignées cellulaires cancéreuses ont été réalisés et les résultats sont présentés dans ce rapport. / Cancer is one of the leading cause of death in France after cardiovascular diseases. Cancer results from an abnormally excessive cell proliferation which leads to the formation of a tumor. The following processes in invasion of tumoral cells and metastasis in the other tissues. Stopping mitosis by chemotherapy can cause death of cancer cells. In order to induce this apoptosis, we are interested by a class of antitumoral drugs which disrupt mitotic spindle function by focusing on its major component: the microtubules. These agents set exactly on their main structural element: the tubulin. The microtubule was recognised as a subcellular target of major strategic importance with regard to anticancer therapeutics. Our work consisted of the design and the synthesis of new antitumor drugs which influence microtubules dynamics. Theses molecules should inhibit tubulin polymerization by binding to the colchicine site. Moreover, we expect theses compounds selectively target tumor vasculature and thus can also be considered vascular disrupting agents. Enzymatic inhibition and cytotoxic assays were performed on different cell lines and are presented in this report.

OBJETIVO: demonstrar as alterações histológicas secundárias ao uso de bevacizumabe na aorta descendente de suínos submetida à interrupção dos vasa vasorum. MATERIAIS E MÉTODOS: em doze suínos, divididos em grupos tratamento e controle (n=6 cada) foi realizada dissecção por 5 cm da aorta torácica, ligadura das artérias intercostais e cobertura com polivinil; o grupo tratamento recebeu uma dose endovenosa única de bevacizumabe. Após quinze dias, os animais foram sacrificados para retirada da artéria e preparo das lâminas histológicas dos grupos tratamento, controle e área não manipulada dos dois grupos. As lâminas foram analisadas com relação aos graus de angiogênese, injúria, inflamação e espessamento intimal. A análise estatística foi conduzida através da média e do desvio-padrão dos escores e as comparações entre os grupos foram realizadas pelo teste de Mann-Whitney. Para obtenção de intervalos de confiança de 95% para as médias das contagens dos escores utilizou-se a distribuição de Poisson, a fim de determinar o efeito estatístico. RESULTADOS: o bevacizumabe causou parefeitos em todos os suínos tratados, com um óbito. As variáveis analisadas através da Escala de Magnitude para Efeito Estatístico, demonstram tendência de redução da angiogênese e da injúria e de aumento da inflamação no limite do moderado. Não ocorreu modificação do espessamento intimal entre os grupos. CONCLUSÃO: a medicação utilizada na lesão da parede arterial induzindo hipóxia, mostrou tendência de redução da angiogênese e da injúria, mas não reduziu o processo inflamatório ou o espessamento intimal da parede arterial. O bevacizumabe mostrou toxicidade no modelo suíno. / OBJECTIVE: To demonstrate histological changes secondary to the use of bevacizumab in the descending aorta of pigs after interruption of vasa vasorum flow. METHODS: Twelve pigs were divided into control and treatment groups (n = 6 each). It was performed a 5-cm dissection of the thoracic aorta, ligation of the intercostal arteries and protection with polyvinyl chloride. Pigs in the treatment group received a single intravenous dose of bevacizumab. After 15 days, the animals were euthanized and the aorta removed. Histological slides were prepared for control and treatment groups and for the non-surgically manipulated part of the aorta in both groups. The slides were analyzed for the degree of angiogenesis, injury, inflammation, and intimal thickening. Data were expressed as mean (SD) of scores and groups were compared using the Mann-Whitney test. The Poisson distribution was used to calculate 95% confidence intervals for the mean scores in order to determine effect statistics. RESULTS: Bevacizumab had adverse effects on all treated pigs, leading to one death. The analysis using a scale of magnitudes for effect statistics showed a trend toward a decrease in angiogenesis and injury and an increase in inflammation for moderate effects. There was no change in intimal thickening in either group. CONCLUSION: The medication used for arterial wall injury inducing hypoxia showed a trend toward reduced angiogenesis and injury, but with no reduction in the inflammatory process or intimal thickening of the arterial wall. Bevacizumab showed toxicity in the porcine model.

OBJETIVO: demonstrar as alterações histológicas secundárias ao uso de bevacizumabe na aorta descendente de suínos submetida à interrupção dos vasa vasorum. MATERIAIS E MÉTODOS: em doze suínos, divididos em grupos tratamento e controle (n=6 cada) foi realizada dissecção por 5 cm da aorta torácica, ligadura das artérias intercostais e cobertura com polivinil; o grupo tratamento recebeu uma dose endovenosa única de bevacizumabe. Após quinze dias, os animais foram sacrificados para retirada da artéria e preparo das lâminas histológicas dos grupos tratamento, controle e área não manipulada dos dois grupos. As lâminas foram analisadas com relação aos graus de angiogênese, injúria, inflamação e espessamento intimal. A análise estatística foi conduzida através da média e do desvio-padrão dos escores e as comparações entre os grupos foram realizadas pelo teste de Mann-Whitney. Para obtenção de intervalos de confiança de 95% para as médias das contagens dos escores utilizou-se a distribuição de Poisson, a fim de determinar o efeito estatístico. RESULTADOS: o bevacizumabe causou parefeitos em todos os suínos tratados, com um óbito. As variáveis analisadas através da Escala de Magnitude para Efeito Estatístico, demonstram tendência de redução da angiogênese e da injúria e de aumento da inflamação no limite do moderado. Não ocorreu modificação do espessamento intimal entre os grupos. CONCLUSÃO: a medicação utilizada na lesão da parede arterial induzindo hipóxia, mostrou tendência de redução da angiogênese e da injúria, mas não reduziu o processo inflamatório ou o espessamento intimal da parede arterial. O bevacizumabe mostrou toxicidade no modelo suíno. / OBJECTIVE: To demonstrate histological changes secondary to the use of bevacizumab in the descending aorta of pigs after interruption of vasa vasorum flow. METHODS: Twelve pigs were divided into control and treatment groups (n = 6 each). It was performed a 5-cm dissection of the thoracic aorta, ligation of the intercostal arteries and protection with polyvinyl chloride. Pigs in the treatment group received a single intravenous dose of bevacizumab. After 15 days, the animals were euthanized and the aorta removed. Histological slides were prepared for control and treatment groups and for the non-surgically manipulated part of the aorta in both groups. The slides were analyzed for the degree of angiogenesis, injury, inflammation, and intimal thickening. Data were expressed as mean (SD) of scores and groups were compared using the Mann-Whitney test. The Poisson distribution was used to calculate 95% confidence intervals for the mean scores in order to determine effect statistics. RESULTS: Bevacizumab had adverse effects on all treated pigs, leading to one death. The analysis using a scale of magnitudes for effect statistics showed a trend toward a decrease in angiogenesis and injury and an increase in inflammation for moderate effects. There was no change in intimal thickening in either group. CONCLUSION: The medication used for arterial wall injury inducing hypoxia showed a trend toward reduced angiogenesis and injury, but with no reduction in the inflammatory process or intimal thickening of the arterial wall. Bevacizumab showed toxicity in the porcine model.

OBJETIVO: demonstrar as alterações histológicas secundárias ao uso de bevacizumabe na aorta descendente de suínos submetida à interrupção dos vasa vasorum. MATERIAIS E MÉTODOS: em doze suínos, divididos em grupos tratamento e controle (n=6 cada) foi realizada dissecção por 5 cm da aorta torácica, ligadura das artérias intercostais e cobertura com polivinil; o grupo tratamento recebeu uma dose endovenosa única de bevacizumabe. Após quinze dias, os animais foram sacrificados para retirada da artéria e preparo das lâminas histológicas dos grupos tratamento, controle e área não manipulada dos dois grupos. As lâminas foram analisadas com relação aos graus de angiogênese, injúria, inflamação e espessamento intimal. A análise estatística foi conduzida através da média e do desvio-padrão dos escores e as comparações entre os grupos foram realizadas pelo teste de Mann-Whitney. Para obtenção de intervalos de confiança de 95% para as médias das contagens dos escores utilizou-se a distribuição de Poisson, a fim de determinar o efeito estatístico. RESULTADOS: o bevacizumabe causou parefeitos em todos os suínos tratados, com um óbito. As variáveis analisadas através da Escala de Magnitude para Efeito Estatístico, demonstram tendência de redução da angiogênese e da injúria e de aumento da inflamação no limite do moderado. Não ocorreu modificação do espessamento intimal entre os grupos. CONCLUSÃO: a medicação utilizada na lesão da parede arterial induzindo hipóxia, mostrou tendência de redução da angiogênese e da injúria, mas não reduziu o processo inflamatório ou o espessamento intimal da parede arterial. O bevacizumabe mostrou toxicidade no modelo suíno. / OBJECTIVE: To demonstrate histological changes secondary to the use of bevacizumab in the descending aorta of pigs after interruption of vasa vasorum flow. METHODS: Twelve pigs were divided into control and treatment groups (n = 6 each). It was performed a 5-cm dissection of the thoracic aorta, ligation of the intercostal arteries and protection with polyvinyl chloride. Pigs in the treatment group received a single intravenous dose of bevacizumab. After 15 days, the animals were euthanized and the aorta removed. Histological slides were prepared for control and treatment groups and for the non-surgically manipulated part of the aorta in both groups. The slides were analyzed for the degree of angiogenesis, injury, inflammation, and intimal thickening. Data were expressed as mean (SD) of scores and groups were compared using the Mann-Whitney test. The Poisson distribution was used to calculate 95% confidence intervals for the mean scores in order to determine effect statistics. RESULTS: Bevacizumab had adverse effects on all treated pigs, leading to one death. The analysis using a scale of magnitudes for effect statistics showed a trend toward a decrease in angiogenesis and injury and an increase in inflammation for moderate effects. There was no change in intimal thickening in either group. CONCLUSION: The medication used for arterial wall injury inducing hypoxia showed a trend toward reduced angiogenesis and injury, but with no reduction in the inflammatory process or intimal thickening of the arterial wall. Bevacizumab showed toxicity in the porcine model.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / FundaÃÃo de Amparo à Pesquisa do Estado do Cearà / Observa-se exacerbaÃÃo do processo angiogÃnico em diversas doenÃas, entre elas pode ser citada a neovascularizaÃÃo de cÃrnea. Para o tratamento dessas doenÃas, sÃo estudados diversos fÃrmacos com propriedades antiangiogÃnicas. O objetivo deste trabalho foi avaliar a eficÃcia do fÃrmaco LASSBIO-596, estruturalmente designado como um hÃbrido da talidomida, sildenafil e arilsulfonamida, na angiogÃnese corneana inflamatÃria. Dezoito coelhos foram submetidos a uma cauterizaÃÃo alcalina pontual na regiÃo superior da cÃrnea direita. Os animais foram randomicamente alocados em trÃs grupos: veÃculo, dexametasona 4,0% e LASSBio-596 1,0%. Os tratamentos foram administrados em forma de colÃrio trÃs vezes por dia durante 21 dias. AvaliaÃÃes foram realizadas nos dias 3, 6, 9, 12, 15, 18 e 21 pÃs-cauterizaÃÃo. Nesses momentos, imagens digitalizadas da cÃrnea foram capturadas de uma forma padronizada. A resposta angiogÃnica foi mensurada utilizando-se um software que foi desenvolvido especificamente para esta finalidade. Foram calculados os seguintes parÃmetros: Ãrea de neovascularizaÃÃo (AN), comprimento vascular total (CT) e nÃmero de vasos sanguÃneos (NV). A partir da AN, calculou-se a taxa de angiogÃnese (TA) em cada grupo estudado. Observou-se que a dexametasona diminuiu significativamente AN, CT e NV durante todas as avaliaÃÃes. Portanto, a dexametasona inibiu completamente a angiogÃnese inflamatÃria da cÃrnea com uma TA de -0,001  0,006 mm2/dia, que foi significativamente menor (p < 0,001) do que a observada apÃs o tratamento com o veÃculo (0,078  0,024 mm2/dia) e LASSBio-596 (0,054  0,012 mm2/dia). Apesar de o LASSBio-596 ter reduzido o processo de angiogÃnese em relaÃÃo ao veÃculo, conforme AN, CT e NV, essa diferenÃa nÃo foi estatisticamente significante. No entanto, verificou-se que a TA medida no grupo LASSBio-596 foi significativamente menor (p < 0,05) do que a observada nos animais controle, indicando um potencial efeito antiangiogÃnico. ConcluÃmos que a aplicaÃÃo tÃpica de LASSBio-596 a 1,0% tem um efeito inibitÃrio parcial sobre a angiogÃnese corneana inflamatÃria em coelhos. / There is exacerbation of the angiogenic process in various diseases, especially in corneal neovascularization. For the treatment of these diseases, several drugs were studied with antiangiogenic properties. The aim of this study was to evaluate the effect of LASSBio-596, structurally designed as a hybrid of thalidomide, sildenafil and arilsulfonamide, on inflammatory corneal angiogenesis. Eighteen rabbits were submitted to an alkaline cauterization in the right cornea. The animals were randomly allocated in three groups: vehicle, dexamethasone, and LASSBio-596 group. Drugs were administrated by eye drops three times per day for 21 days. Evaluations were performed on days 3, 6, 9, 12, 15, 18, and 21 post-cauterization. At these time points, digital images of the cornea were captured in a standard fashion. Angiogenic response was measured using software that was developed specifically for this purpose. It calculated the following parameters: neovascularization area (NA), total vascular length (TVL), and blood vessel number (BVN). It was observed that dexamethasone significantly decreased NA, TVL, and BVN during all assessments. From NA it was calculated the angiogenesis rate (AR) in each group. Therefore, dexamethasone completely inhibited the inflammatory corneal angiogenesis with angiogenesis rate (AR) of -0.001  0.006 mm2/day, which was significantly lower (p < 0.001) than that observed after treatment with vehicle (0.078  0.024 mm2/dia) and LASSBio-596 (0.054  0.012 mm2/dia). Although LASSBio-596 reduced angiogenesis in relation to vehicle, according to NA, TVL and BVN values, this difference was not statistically significant. However, it was found that the AR as measured in the LASSBio-596 group was significantly lower (p < 0.05) than that seen in control animals, indicating a potential antiangiogenic effect. We conclude that topical application of LASSBio-596 at 1.0% has a partial inhibitory effect on inflammatory corneal angiogenesis in rabbits.