SYNTHETIC LIPID-A WITH ENDOTOXIC AND RELATED BIOLOGICAL-ACTIVITIES COMPARABLE TO THOSE OF A NATURAL LIPID-A FROM AN ESCHERICHIA-COLI RE-MUTANT
SYNTHETIC LIPID-A WITH ENDOTOXIC AND RELATED BIOLOGICAL-ACTIVITIES COMPARABLE TO THOSE OF A NATURAL LIPID-A FROM AN ESCHERICHIA-COLI RE-MUTANT
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DOI：
10.1128/iai.49.1.225-237.1985
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发表时间：
1985-01-01
影响因子：
3.1
通讯作者：
YOSHIMURA, H
中科院分区：
文献类型：
--
作者：
KOTANI, S;TAKADA, H;YOSHIMURA, H
一种合成化合物（506），(1-6)D-葡糖胺二糖1，4 ″-二磷酸，其在2 ″-氨基和3 ″-羟基分别被（R）-3-十二烷酰氧基十四烷酰基和（R）-3-十四烷酰氧基十四烷酰基酰化，并且在2-氨基和3-羟基具有（R）-3-羟基十四烷酰基，表现出与来自大肠杆菌再突变体（菌株F515）的参比脂质A相同或有时更强的完全内毒素活性。测试的内毒素活性包括家兔致热原性和白细胞减少诱导活性、正常小鼠体重降低毒性、半乳糖胺致敏小鼠和鸡胚致死毒性以及家兔局部Shwartzman反应的制备和激发。相比之下，化合物406（脂质A分子的生物合成前体的合成对应物）在所有上述测定系统中仅显示出弱活性，除了在半乳糖胺负载的小鼠中的致死性。这一发现有力地表明，在二糖骨架的C-2“和C-3”位置上存在酰氧基酰基是脂质A分子的最重要的决定性结构之一，其表现出脂多糖及其脂质A部分的强生物活性特征。相应的4“-单磷酸酯（化合物504）和1-单磷酸酯（505）类似物的活性显著低于母体分子506和参比F515脂质A的活性。关于其他生物活性，不仅化合物506，而且化合物504、505和406也显示出确定的活性，有时与F515脂质A和其他参考天然产物的活性相当。这些是通过经典途径激活中国鲎变形细胞凝血酶级联和人补体、小鼠脾细胞的促有丝分裂和多克隆B细胞激活、豚鼠腹腔巨噬细胞刺激、人血液多形核白细胞迁移增强以及诱导牛分枝杆菌BCG致敏小鼠中对L-929细胞具有细胞抑制和细胞杀伤作用的血清因子。供试合成化合物的相对效价取决于测定系统，并且在不同系统之间存在差异。脱磷酸化合物503缺乏在磷酸化化合物中明确观察到的大部分生物活性，可能是因为其不溶性。本研究表明，成功的化学合成的E。大肠杆菌型脂质A.
A synthetic compound (506), .beta.(1-6) D-glucosamine disaccharide 1,4''-bisphosphate, which is acylated at 2''-amino and 3''-hydroxyl groups with (R)-3-dodecanoyloxytetradecanoyl and (R)-3-tetradecanoyloxytetradecanoyl groups, respectively, and has (R)-3-hydroxytetradecanoyl groups at 2-amino and 3-hydroxyl groups, exhibited full endotoxic activities identical or sometimes stronger than those of a reference lipid A from an Escherichia coli Re-mutant (strain F515). Endotoxic activities tested include pyrogenicity and leukopenia-inducing activity in rabbits, body weight-decreasing toxicity in normal mice, lethal toxicity in galactosamine-sensitized mice and chicken embyros, and the preparation and provocation of the local Shwartzman reaction in rabbits. Compound 406, a synthetic counterpart of a biosynthetic precursor of lipid A molecule, showed by contrast only weak activities in all of the above assay systems except for the lethality in galactosamine-loaded mice. This finding strongly suggests that the presence of acyloxyacyl groups at the C-2'' and C-3'' positions of the disaccharide backbone is one of the most important determinant structures of the lipid A molecule for exhibition of strong biological activities characteristic of lipopolysaccharide and its lipid A moiety. The activities of the corresponding 4''-monophosphate (compound 504) and 1-monophosphate (505) analogs were considerably less than those of the parent molecule 506 and the reference F515 lipid A. Regarding other biological activities, not only compound 506 but also compounds 504, 505, and 406 showed definite activities, sometimes comparable to those of F515 lipid A and other reference natural products. These are the activation of Tachypleus tridentatus amoebocyte clotting enzyme cascade and human complement via the classical pathway, mitogenic and polyclonal B-cell activation of murine splenocytes, stimulation of peritoneal macrophages in a guinea pig, enhancement of migration of human blood polymorphonuclear leukocytes, and induction of a serum factor that is cytostatic and cytocidal to L-929 cells in Mycobacterium bovis BCG-primed mice. Relative potencies of test synthetic compounds depended on the assay systems and varied from one system to another. Dephospho-compound 503 lacked most of the biological activities that were definitely observed with phosphorylated compounds, probably because of its insolubility. This study demonstrates the successful chemical synthesis of an E. coli-type lipid A.