Wai Kei Chan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 155-164). / Abstracts in English and Chinese. / Chapter Chapter 1 --- Introduction / Chapter 1.1. --- Sulfonylureas --- p.1 / Chapter 1.2. --- Biological action of sulfonylurea drugs --- p.2 / Chapter 1.2.1 --- Effects on pancreatic β cells --- p.5 / Chapter 1.2.2. --- Effects on cardiac myocytes --- p.7 / Chapter 1.2.3. --- Effects on smooth muscle cells --- p.11 / Chapter 1.2.4. --- Effects on endothelial cells --- p.14 / Chapter 1.3. --- Side effects and toxicity --- p.15 / Chapter 1.4. --- Objectives of the present study --- p.17 / Chapter Chapter 2 --- Methods and Marterials / Chapter 2.1. --- Tissue and Cell Preparation --- p.20 / Chapter 2.1.1. --- Preparation of the isolated rat aorta and mesenteric artery --- p.20 / Chapter 2.1.2. --- Removal of the functional endothelium --- p.20 / Chapter 2.1.3. --- Cell culture --- p.21 / Chapter 2.1.3.1. --- Materials --- p.21 / Chapter 2.1.3.2. --- Aortic smooth muscle cells in primary culture --- p.21 / Chapter 2.1.3.3. --- Aortic endothelial cells in primary culture --- p.23 / Chapter 2.1.3.4. --- Cultured rat aortic smooth muscle cell line (A7r5) --- p.23 / Chapter 2.1.3.5. --- Cultured human umbilical vein endothelial cell line (ECV-304) --- p.24 / Chapter 2.1.3.6. --- Cell subculture --- p.24 / Chapter 2.1.3.7. --- Immunostaining of endothelial cells in primary culture --- p.24 / Chapter 2.2. --- Organ Bath Set-up --- p.25 / Chapter 2.3. --- Force Measurement --- p.28 / Chapter 2.3.1. --- Vascular action of glibenclamide --- p.28 / Chapter 2.3.1.1. --- Antagonistic effect of glibenclamide on relaxation induced by K+ channel openers --- p.28 / Chapter 2.3.1.2. --- Relaxant response of glibenclamide --- p.29 / Chapter 2.3.1.3. --- Role of endothelium-derived vasoactive factors in glibenclamide induced relaxation --- p.29 / Chapter 2.3.1.4. --- Effect of endothelial prostanoids in glibenclamide-induced relaxation --- p.30 / Chapter 2.3.1.5. --- Effects of putative K+ channel blockers on glibenclamide-induced relaxation --- p.30 / Chapter 2.3.1.6. --- Effect of glibenclamide on high K+- and CaCl2-induced contraction --- p.31 / Chapter 2.3.1.7. --- Effect of glibenclamide on prostaglandin F2α-induced contraction --- p.32 / Chapter 2.3.1.8. --- Effect of glibenclamide on protein kinase C-mediated contraction --- p.32 / Chapter 2.3.2. --- Vascular action of glipizide --- p.33 / Chapter 2.3.3. --- Vascular action of tolbutamide --- p.33 / Chapter 2.3.3.1. --- Contractile response of tolbutamide --- p.33 / Chapter 2.3.3.2. --- Effects of inhibitors of endothelium-derived factors --- p.33 / Chapter 2.3.3.3. --- Effects of inhibitors of Ca2+ influx --- p.34 / Chapter 2.3.3.4. --- Effect of protein kinase C inhibitor --- p.34 / Chapter 2.3.3.5. --- Effects of neural factors --- p.34 / Chapter 2.4. --- Cyclic GMP measurement --- p.35 / Chapter 2.4.1. --- Material --- p.35 / Chapter 2.4.2. --- Methods --- p.35 / Chapter 2.4.2.1. --- Tissue preparation --- p.35 / Chapter 2.4.2.2. --- Plasma and tissue according to protocols provided by Amersham --- p.35 / Chapter 2.4.2.3. --- Cyclic GMP content measurement --- p.36 / Chapter 2.4.2.4. --- Protein content measurement --- p.39 / Chapter 2.4.2.5. --- Cyclic GMP measurement protocol --- p.40 / Chapter 2.5. --- Ca2+ measurement --- p.40 / Chapter 2.5.1. --- Materials --- p.40 / Chapter 2.5.1.1. --- PTI RatioMaster Fluorescence System --- p.40 / Chapter 2.5.1.2. --- Confocal Imaging System --- p.42 / Chapter 2.5.2. --- Method --- p.42 / Chapter 2.5.3. --- Protocols for Ca2+ measurement --- p.45 / Chapter 2.5.3.1. --- Effect of glibenclamide in endothelial cells --- p.45 / Chapter 2.5.3.2. --- Effect of glibenclamide in vascular smooth muscle cells --- p.45 / Chapter 2.5.3.3. --- Effect of tolbutamide in vascular smooth muscle cells --- p.46 / Chapter 2.6. --- Cell proliferation --- p.45 / Chapter 2.6.1. --- Materials --- p.45 / Chapter 2.6.2. --- Method --- p.46 / Chapter 2.6.3. --- Protocols for cell proliferation --- p.47 / Chapter 2.6.3.1. --- Effect of glibenclamide on endothelial cell proliferation --- p.47 / Chapter 2.6.3.2. --- Effect of glibenclamide on aortic smooth muscle cell proliferation --- p.47 / Chapter 2.7. --- Chemicals and solutions --- p.48 / Chapter 2.8. --- Statistical analysis --- p.50 / Chapter Chapter 3 --- Results / Chapter 3.1. --- Glibenclamide --- p.51 / Chapter 3.1.1. --- Effect of glibenclamide on the K+ channel activity --- p.51 / Chapter 3.1.2. --- Relaxant response of glibenclamide --- p.55 / Chapter 3.1.3. --- Effects of inhibitors of nitric oxide activity on glibenclamide- induced relaxation --- p.57 / Chapter 3.1.4. --- Role of endothelial relaxing prostanoids in glibenclamide-induced relaxation --- p.69 / Chapter 3.1.5. --- Effect of putative K+ channel blockers on glibenclamide-induced relaxation --- p.73 / Chapter 3.1.6. --- Effect of glibenclamide on high K+-induced arterial contraction --- p.75 / Chapter 3.1.7. --- Effect of glibenclamide on protein kinase C-mediated contraction --- p.83 / Chapter 3.1.8. --- Effect of glibenclamide on prostaglandin F2α-induced contraction --- p.83 / Chapter 3.2 --- Glipizide --- p.85 / Chapter 3.2.1. --- Relaxant response of glipizide --- p.85 / Chapter 3.3. --- Tolbutamide --- p.91 / Chapter 3.3.1. --- Contractile response to tolbutamide --- p.91 / Chapter 3.3.2. --- Effects of endothelium-derived factors --- p.94 / Chapter 3.3.3. --- Effects of inhibitors of Ca2+ influx on tolbutamide-induced contraction --- p.98 / Chapter 3.3.4. --- "Effects of forskolin, sodium nitroprusside, staurosporine on tolbutamide-induced contraction" --- p.102 / Chapter 3.3.5. --- Effect of neural factors --- p.106 / Chapter 3.4. --- Effect of glibenclamide on cGMP levels --- p.112 / Chapter 3.5. --- Effect of glibenclamide on intracellular[Ca2+ ] in cultured endothelial cells --- p.112 / Chapter 3.6. --- Effect of glibenclamide on intracellular [Ca2+] in cultured aortic smooth muscle cells --- p.115 / Chapter 3.7. --- Effect of tolbutamide on intracellular [Ca2+] in cultured aortic smooth muscle cells --- p.121 / Chapter 3.8. --- Effect of glibenclamide on proliferation of cultured endothelial cells --- p.121 / Chapter 3.9. --- Effect of glibenclamide on proliferation of cultured aortic smooth muscle cells --- p.123 / Chapter Chapter 4 --- Discussion / Chapter 4.1. --- Effect of glibenclamide --- p.133 / Chapter 4.2. --- Effect of glipizide --- p.143 / Chapter 4.3. --- Effect of tolbutamide --- p.144 / Chapter 4.4. --- Conclusion --- p.152 / References --- p.155 / Publications --- p.163

A tolerância da cultura de milho aos herbicidas do grupo das sulfoniluréias, aplicados em condições de pós-emergência é variável em função do híbrido cultivado. Da mesma forma, tem sido observada tolerância diferencial ao nicosulfuron entre as cultivares de arroz melhoradas para tolerar os herbicidas imazethapyr + imazapic (AHAS-tolerante). Desta forma, este trabalho foi desenvolvido com o objetivo de avaliar: a seletividade do herbicida nicosulfuron, aplicado em condições de pós-emergência, a híbridos de milho e cultivares de arroz AHAS-tolerante. No experimento com a cultura do milho, foram estudados dez híbridos designados comercialmente por DKB 370, DKB 990, AG 6020, AG 9040, AS 1551, AS 1572, SWB 585, BX 1149, BM 620 e BM 128, sendo o delineamento experimental adotado em blocos casualizados, com cinco tratamentos e quatro repetições. Os tratamentos utilizados em g i.a. ha-1 foram: nicosulfuron + atrazina a 20 + 1500; nicosulfuron + atrazina a 40 + 3000; nicosulfuron a 50; nicosulfuron a 60, bem como a testemunha capinada, aplicados no estádio fenológico de quatros folhas expandidas (V4). Os híbridos que foram tolerantes a todos os tratamentos, não apresentando reduções de produção, foram DKB 370, AG 9040, AS 1551 e BM 620. Os tratamentos de nicosulfuron em mistura com atrazina nas doses de 20 + 1500 e 40 + 3000 g ha-1 de i.a. foram seletivos para todos os híbridos testados. As doses de nicosulfuron a 50 e 60 g i a ha-1 foram as mais fitotóxicas do experimento, os híbridos DKB 990, AG 6020, AS 1572, SWB 585, BX 1149 e BM 128 apresentaram redução de produção para ambas as dose. Para os híbridos AG 6020 e SWB 585 ocorreu diferenças de producão entre os tratamentos de nicosulfuron a 50 e 60 g i a ha- 1, o que indica suscetibilidade ao nicosulfuron. A tolerância ao nicosulfuron das duas cultivares de arroz AHAS-tolerante IRGA 422 CL e Puitá CL INTA foi testada em um outro experimento. Os tratamentos resultaram de esquema do tipo fatorial 2 x 8, em que duas foram as cultivares de arroz e oito foram as doses de nicosulfuron (0; 6,25; 12,5; 25; 50; 100; 200 e 400 g i.a.ha-1). Foram realizadas avaliações visuais de sintomas de fitotoxicidade aos 15 e 20 dias após a aplicação (DAA) e aos 20 DAA avaliou-se o comprimento de raiz, comprimento da parte aérea das plantas e massa seca de cada cultivar separadamente. Os dados foram submetidos à aplicação do teste F na análise da variância posteriormente a aplicação de regressões não lineares do tipo log-logístico. A cultivar IRGA 422 CL tolerou maiores doses de nicosulfuron quando comparada com a cultivar Puitá CL INTA. / The tolerance of the crop corn to the sulfonylurea herbicides, sprayed in post emergence conditions, is variable according to the hybrids that is cultivated. Likewise, it has been observed differential tolerance to nicosulfuron among rice cultivars bred to tolerate the herbicides imazethapyr + imazapic (AHAS-tolerant). Therefore, this research was developed in order to evaluate the selectivity of the herbicide nicosulfuron, sprayed in post emergence conditions, to corn hybrids and AHAS-tolerant rice cultivars. In the experiment with the crop corn, it was studied ten hybrids commercially designated as DKB 370, DKB 990, AG 6020, AG 9040, AS 1551, AS 1572, SWB 585, BX 1149, BM 620 e BM 128, being the experimental design adopted romdomized completely blocks, with five treatments and four replications. The treatments used in g a.i. ha-1 were: nicosulfuron + atrazine at 20 + 1,500; nicosulfuron + atrazine at 40 + 3,000; nicosulfuron at 50; nicosulfuron at 60, as well as a check weed free, sprayed in the phenological stage of four expanded leaves (V4). The hybrids that were tolerant to all treatments, with no yield reduction, were DKB 370, AG 9040, AS 1551 e BM 620. The treatments of nicosulfuron in mixture with atrazine at the rates of 20 + 1,500 e 40 + 3,000 g ha-1 of i.a. were selective to all hybrids tested. The rates of nicosulfuron at 50 and 60 g a.i. ha-1 were the least selective of the experiment, the hybrids DKB 990, AG 6020, AS 1572, SWB 585, BX 1149 and BM 128 showed yield reduction. For the hybrids AG 6020 and SWB 585 there were diferences in the yield among treatments of nicosulfuron 50 and 60 g a.i. ha-1, indicating low tolerance to nicosulfuron. The tolerance to nicosulfuron of the AHAS-tolerant rice cultivars IRGA 422 CL and Puita CL INTA was tested in another experiment. The treatments were obtained from a factorial design 2 x 8, with two rice cultivars and eight rates of nicosulfuron (0, 6.25, 12.5, 25, 50, 100, 200 and 400 g a.i.ha- 1). Visual observations of phytotoxicity symptoms were evaluated at 15 and 20 days after treatments (DAA) and at 20 DAA it was evaluated the root and shoot length and dry biomass of each cultivar separately. The data were submitted to F test in the analysis of variance followed by the application of non linear regressions log-logistic type. The cultivar IRGA 422 CL tolerated higher rates of nicosulfuron when compared to the cultivar Puita CL Inta.

SUR2A is a member of the ABC transporter superfamily. SUR2A mediated regulation of KATP channels is essential as mutations in the nucleotide binding domains (NBDs) of SUR2A are associated with cardiovascular disorders. Studies of eukaryotic NBDs, such as SUR2A, are hindered by low solubility of the isolated domain. We hypothesized that the solubility of heterologously expressed SUR2A NBDs depends on the definition of the domain boundaries. Boundaries were initially predicted using a combination of a structure-based sequence alignment and homology modeling, and subsequently verified by testing the solubility of five SUR2A NBD1 constructs with different N- or C-terminal boundaries. The boundaries of SUR2A NBD1 essential for solubility were identified. CD and NMR data indicate that SUR2A NBD1 is folded. Our method may be applied as a general method for developing suitable constructs of other NBDs of ABC proteins such as SUR isoforms, SUR2B and SUR2C, and the vacuolar transporter, Ycf1p.

ATP-sensitive potassium (KATP) channels have crucial roles in several biological processes. KATP channels possess four regulatory sulfonylurea receptors. The SUR proteins are members of the ubiquitous ATP-binding cassette (ABC) superfamily. However, unlike most ABC proteins, SURs do not transport substrates but function strictly as regulators of KATP channel activity. Currently, studies into the molecular basis by which various mutations in SUR2A cause disease are highly limited. This is primarily a consequence of poor solubility of isolated SUR2A NBDs, as is typical for many eukaryotic NBDs. By employing structure-based sequence alignments and biophysical studies, we determined domain boundaries for SUR2A NBD1 that enabled, for the first time, NMR studies of NBD1. Our biophysical studies demonstrate that the isolated SUR2A NBD1 is folded and exhibits differential dynamics upon ATP binding activity. Additional studies are now possible to examine the effects of disease-causing mutations on structure, dynamics, and interactions of NBD1.

SUR2A is a member of the ABC transporter superfamily. SUR2A mediated regulation of KATP channels is essential as mutations in the nucleotide binding domains (NBDs) of SUR2A are associated with cardiovascular disorders. Studies of eukaryotic NBDs, such as SUR2A, are hindered by low solubility of the isolated domain. We hypothesized that the solubility of heterologously expressed SUR2A NBDs depends on the definition of the domain boundaries. Boundaries were initially predicted using a combination of a structure-based sequence alignment and homology modeling, and subsequently verified by testing the solubility of five SUR2A NBD1 constructs with different N- or C-terminal boundaries. The boundaries of SUR2A NBD1 essential for solubility were identified. CD and NMR data indicate that SUR2A NBD1 is folded. Our method may be applied as a general method for developing suitable constructs of other NBDs of ABC proteins such as SUR isoforms, SUR2B and SUR2C, and the vacuolar transporter, Ycf1p.

ATP-sensitive potassium (KATP) channels have crucial roles in several biological processes. KATP channels possess four regulatory sulfonylurea receptors. The SUR proteins are members of the ubiquitous ATP-binding cassette (ABC) superfamily. However, unlike most ABC proteins, SURs do not transport substrates but function strictly as regulators of KATP channel activity. Currently, studies into the molecular basis by which various mutations in SUR2A cause disease are highly limited. This is primarily a consequence of poor solubility of isolated SUR2A NBDs, as is typical for many eukaryotic NBDs. By employing structure-based sequence alignments and biophysical studies, we determined domain boundaries for SUR2A NBD1 that enabled, for the first time, NMR studies of NBD1. Our biophysical studies demonstrate that the isolated SUR2A NBD1 is folded and exhibits differential dynamics upon ATP binding activity. Additional studies are now possible to examine the effects of disease-causing mutations on structure, dynamics, and interactions of NBD1.

KATP channels couple cell metabolic status to the membrane excitability by sensing the cytoplasmic ATP/ADP ratio. Present studies examined how conserved motifs (Walker A (WA), signature sequence (L), and Walker B (WB)) within each NBF of SUR2A bind to Syn-1A to affect its actions on cardiac KATP channels. In vitro binding experiments illustrated that Syn-1A binds cardiac SUR2A at WA and L of NBF-1 and WA, L, and WB of NBF-2. Electrophysiology experiments on stably expressing SUR2A/Kir6.2 cell-lines showed that only L and WB of NBF-1 and all three NBF-2 motifs could abrogate the inhibitory effect of Syn-1A on SUR2A/KATP channels. These results lead me to hypothesize that more independent motif in NBF-2 can bind and abrogate Syn-1A’s inhibition than NBF-1 on SUR2A/KATP channels. A corollary postulate is that Syn-1A acts as a scaffold to secure the NBF-1 and -2 in dimer conformation required for SUR2A to modulate Kir6.2 gating.

KATP channels couple cell metabolic status to the membrane excitability by sensing the cytoplasmic ATP/ADP ratio. Present studies examined how conserved motifs (Walker A (WA), signature sequence (L), and Walker B (WB)) within each NBF of SUR2A bind to Syn-1A to affect its actions on cardiac KATP channels. In vitro binding experiments illustrated that Syn-1A binds cardiac SUR2A at WA and L of NBF-1 and WA, L, and WB of NBF-2. Electrophysiology experiments on stably expressing SUR2A/Kir6.2 cell-lines showed that only L and WB of NBF-1 and all three NBF-2 motifs could abrogate the inhibitory effect of Syn-1A on SUR2A/KATP channels. These results lead me to hypothesize that more independent motif in NBF-2 can bind and abrogate Syn-1A’s inhibition than NBF-1 on SUR2A/KATP channels. A corollary postulate is that Syn-1A acts as a scaffold to secure the NBF-1 and -2 in dimer conformation required for SUR2A to modulate Kir6.2 gating.

Objective: To describe clinical presentation/longterm outcomes of patients with ABCC8/KCNJ11 variants in a large cohort of patients with diabetes. Research Design and Methods: We analyzed patients in the Diabetes Prospective Follow-up (DPV) registry with diabetes and pathogenic variants in the ABCC8/ KCNJ11 genes. For patients with available data at three specific time-points— classification as K+-channel variant, 2-year follow-up and most recent visit—the longitudinal course was evaluated in addition to the cross-sectional examination. Results: We identified 93 cases with ABCC8 (n = 54)/KCNJ11 (n = 39) variants, 63 of them with neonatal diabetes. For 22 patients, follow-up data were available. Of these, 19 were treated with insulin at diagnosis, and the majority of patients was switched to sulfonylurea thereafter. However, insulin was still administered in six patients at the most recent visit. Patients were in good metabolic control with a median (IQR) A1c level of 6.0% (5.5–6.7), that is, 42.1 (36.6–49.7) mmol/mol after 2 years and 6.7% (6.0– 8.0), that is, 49.7 (42.1–63.9) mmol/mol at the most recent visit. Five patients were temporarily without medication for a median (IQR) time of 4.0 (3.5–4.4) years, while two other patients continue to be off medication at the last follow-up. Conclusions: ABCC8/KCNJ11 variants should be suspected in children diagnosed with diabetes below the age of 6 months, as a high percentage can be switched from insulin to oral antidiabetic drugs. Thirty patients with diabetes due to pathogenic variants of ABCC8 or KCNJ11 were diagnosed beyond the neonatal period. Patients maintain good metabolic control even after a diabetes duration of up to 11 years

Mundialmente, a soja é considerada uma das principais fontes de produção de óleos e proteínas vegetais para alimentação humana e animal. Constitui-se atualmente um dos produtos de maior importância na economia brasileira. É notório o crescimento das áreas ocupadas pelas lavouras de soja no Brasil, que na safra 2014/2015 atingiram 32,09 milhões de hectares, com previsão de expansão para 32,20 milhões de hectares para a safra 2015/2016. Outro aspecto a ser salientado é que as lavouras de soja RR (Roundup Ready) corresponderam a 93,5%, da área total cultivada com soja no Brasil na safra 2014/2015. Cultivares STS (Soja Tolerante à Sulfoniluréias) contém um gene que aumenta a degradação de alguns herbicidas na planta, como por exemplo, o chlorimuron-ethyl, para o qual são posicionados tolerando doses até quatro vezes as doses recomendadas para cultivares não tolerantes, sem apresentar danos significativos. A soja STS foi desenvolvida através da técnica de mutagênese de sementes utilizando o agente alquilante etilmetanosulfonato (EMS), não sendo uma cultura transgênica. Os objetivos do presente trabalho foram: avaliar a seletividade do herbicida chlorimuron-ethyl aplicado em pós-emergência de soja RR/STS; avaliar a seletividade de herbicidas inibidores da ALS aplicados em pós-emergência de soja RR/STS. Para tanto foram conduzidos dois experimentos principais. O primeiro conduzido durante quatro safras (2011/2012 a 2014/2015), os tratamentos foram constituídos por sete doses do herbicida chlorimuron-ethyl (0, 15, 30, 45, 60, 75 e 90 g i. a. ha-1). Foi realizada avaliação das variáveis relacionadas ao desempenho agronômico (altura, número de vagens por planta, produtividade, e massa de mil sementes). O segundo durante a safra 2014/2015 no campo e casa de vegetação, em que a soja RR/STS foi submetida à aplicação de herbicidas inibidores da ALS, associados ou não ao glyphosate. Foi realizada avaliação de fitointoxicação, índice SPAD, bem como variáveis relacionadas ao desempenho agronômico para o experimento no campo (número de vagens por planta e produtividade). Os cultivares de soja CD 250 e CD 2630 RR/STS apresentaram-se tolerantes para aplicação, em pós-emergência (V4), do herbicida chlorimuron-ethyl até a dose de 90 g i. a. ha-1. O cultivar de soja CD 2630 RR/STS apresentou-se tolerante à aplicação, em pós-emergência, dos herbicidas utilizados, isolada ou em associação com glyphosate. Exceção feita ao metsulfuron-methy (2,4 g i. a. ha-1), associado ou não com glyphosate (960 g e. a. ha-1). / Worldwide soybean is considered one of the major sources of production of vegetable oils and proteins for food and feed. It currently is one of the most important products in the Brazilian economy. It is notable growth areas occupied by soybean crops in Brazil, that in the 2014/2015 season reached 32.09 million hectares, with expansion forecast to 32.20 million hectares for the season 2015/2016. Another aspect to be noted is that the RR soybean (Roundup Ready) crops corresponded to 93.5% of the total area cultivated with soybeans in Brazil in the 2014/2015 season. Cultivars STS (Sulfonylurea Tolerant Soybeans) contains a gene that increases the degradation of some herbicides in the plant, for example, chlorimuron-ethyl, to which are positioned tolerated doses up to four times the recommended dosages for nontolerant cultivars, without presenting significant damage. STS soybean was developed by seed mutagenesis technique using the alkylating agent ethyl methane sulfonate (EMS), not being a transgenic crop. The aims of this study were to evaluate the selectivity of the herbicide chlorimuron-ethyl applied post-emergence of RR/STS soybean; evaluate the selectivity of ALS-inhibiting herbicides applied postemergence of RR/STS soybean. Therefore, two main experiments were conducted. The first conducted for four seasons (2011/2012 to 2014/2015), the treatments were seven doses of the herbicide chlorimuron-ethyl (0, 15, 30, 45, 60, 75 and 90 g a. i. ha-1). Performed an assessment of variables related to agronomic performance (height, number of pods per plant, yield and mass of one thousand seeds). The second during the 2014/2014 season in the field and the greenhouse, where RR/STS soybean was subjected to application of ALS inhibitor herbicides, with or without glyphosate. It was conducted evaluation phytointoxication, SPAD index, as well as variables related to agronomic performance (number of pods per plant and productivity). The soybean cultivars CD 250 and CD 2630 RR/STS were tolerant to application, post-emergence (V4) of herbicide chlorimuron-ethyl up to a dose of 90 g a. i. ha-1. Cultivar CD 2630 RR/STS presented tolerance to application in postemergence herbicides used alone or in combination with glyphosate. Except for the metsulfuron-methy (2.4 g a. i. ha-1), associated or not with glyphosate (960 g a. e. ha-1).